O tratamento cirúrgico por câncer colorretal (CCR) traz consequências fisiológicas e psicossociais na vida dos pacientes e de seus familiares, cujos desafios estão vinculados ao estigma social do câncer e as repercussões mutilatórias. Este estudo teve por objetivos analisar a produção científica nacional e internacional sobre a assistência de enfermagem perioperatória para pacientes com câncer colorretal e estabelecer as recomendações para a sistematização da assistência perioperatória com vistas à alta hospitalar. Trata-se de uma Revisão Integrativa, fundamentada na Prática Baseada em Evidências, cuja pergunta formulada foi: Quais as evidências científicas sobre a alta hospitalar do paciente cirúrgico com câncer colorretal? Os descritores utilizados para as buscas foram Colorectal Neoplasms, Nursing, Discharge, nas bases de indexação eletrônica PubMed, Web of Sciences, Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Scopus, Cumulative Index of Nursing and Allied Health Literature (Cinahl) e BDEnf , mediante os critérios de inclusão: estudos primários que abordassem a alta hospitalar do paciente cirúrgico adulto ou idoso com câncer colorretal; publicados no período entre 2005 e 2015 em inglês, espanhol e português; obtidos na íntegra via Biblioteca Central do Campus de Ribeirão Preto da Universidade de São Paulo; e os critérios de exclusão que foram estudos de revisão de literatura e em formato de monografia, dissertação ou tese. Do total de 47 artigos científicos, resultantes das buscas, foram selecionados mediante os critérios de inclusão e exclusão, sete (7) estudos que constituíram a amostra final, sendo que todos foram publicados em inglês, com níveis de evidência pouco fortes (VI e VII) e cujos aspectos abordados foram categorizados em três temáticas: Conhecimentos necessários aos enfermeiros e habilidades requeridas para a sistematização da alta hospitalar; Estratégias para implementar alta hospitalar e Necessidades de pacientes com CCR e familiares. Na temática Conhecimentos e habilidades para o planejamento da alta hospitalar, evidenciou-se que aspectos como fisiopatologia, tratamentos e suas consequências, prognósticos e experiência clínica são importantes para oferecer o suporte profissional adequado às demandas de necessidades de pacientes com CCR e família no perioperatório e para prepará-los para o contexto domiciliário. Na temática Estratégias para implementar alta hospitalar foi enfatizado a utilização do Processo de Enfermagem, de materiais educativos e cuidados especializados, com focalização de conhecimentos sobre a fisiopatologia oncológica, tratamentos e suas consequências e aspectos clínicos para atender a demanda de cuidados especializados desta clientela no perioperatório. Na temática Necessidades de pacientes com CCR e seus familiares verificamos que a demanda de intervenções de enfermagem são para prevenção de infecção do sítio cirúrgico, melhoria do catabolismo e da aprendizagem sobre fisiopatologia oncológica, tratamentos e suas consequências, além do seguimento de controle oncológico, com inclusão da família. Acreditamos que os resultados deste estudo possam subsidiar o planejamento da assistência de enfermagem perioperatória aos pacientes com câncer colorretal para o atendimento da demanda de necessidades específicas do paciente e familiar, favorecendo a transição hospital domicílio / The surgical treatment for colorectal cancer (CRC) brings physiological and psychosocial consequences in patients and their relative\'s life, whose challenges are linked to social stigma of cancer and the repercussions of mutilation. This study aimed to analyze the national and international scientific production about the nursing perioperative assistance to patients with colorectal cancer and establish the recommendations to the systematization of the nursing perioperative assistance aiming the discharge. This is a Integrative Review, grounded in the Evidence Based Practice, whose question formulated was: Which scientific evidences about colorectal cancer patient discharge? The keywords used to search was Colorectal Neoplasms, Nursing, Discharge, at the electronic indexed databases PubMed, Web of Sciences, Literature in the Health Sciences in Latin America and the Caribbean (Lilacs), Scopus, Cumulative Index of Nursing and Allied Health Literature (Cinahl) and BDEnf, by the inclusion criteria: original research about the discharge of the adult or aged patient with colorectal cancer; published between 2005 and 2015 in English, Spanish and Portuguese; with full text by Central Library USP Ribeirão Preto; and the exclusion criteria was reviews, monographs, dissertations and theses. Of 47 scientific papers, result of search, were selected by inclusion and exclusion criteria, seven (7) papers that constituted the final sample, all in English, with little strong reviewed evidence (VI and VII) and whose aspects were categorized in three theme: Knowledge required to nurses and required skills to systematize the discharge; Strategies for implement the discharge and Needs of patients with colorectal cancer and relatives. At Knowledge required to nurses and required skills to systematize the discharge, it became clear that aspects like pathophysiology, treatments and their consequences, prognosis and clinical experience are important to offer professional support suitable for the demands of needs of patients with colorectal cancer and their family at perioperative and to prepare them to home context. At Strategies for implement the discharge it became clear that the utilization of Nursing Process, of specialized educational material, with focusing about the oncologic pathophysiology, treatments and their consequences and clinical aspects to meet the demand of specialized care this clientele at perioperative. At Needs of patients with CRC and relatives we checked that the demand of nursing interventions are to prevent the surgical site infections, to improve catabolism and about the learning of oncologic pathophysiology, treatments and their consequences, beyond to oncologic control, with family including. We believe that the results of this study can subsidize the planning of nursing perioperative care to patients with CRC to meet the demand of specific to patients and their relatives, favoring the transition to hospital for home

大腸癌是全世界最常見的癌症之一，亦是一個癌症死亡率的首要原因。大腸癌患者約50%在病程中會出現轉移病灶。近十年來，雖然多種被批准用於臨床治療的新化療藥顯著提高了大腸癌的治療效果，但是轉移性大腸癌病人的預後仍然很差。隨著各種分子生物技術的進步，新的治療標靶可能在大腸癌細胞株中被發現，並得以在病人標本中驗證。 / 在本研究中，我們採用即時定量多聚酶鏈反應(qPCR)陣列分析，比較大腸癌細胞株和正常大腸細胞株基因表達譜，試圖識別潛在的新的治療標靶。結果提示，與正常大腸細胞株 CCD-18Co 比較，Jagged 2 (JAG2) 和 Frizzled-3 (FZD3)基因 在大腸癌細胞株 SW480 和 SW620 中表達升高。病人大腸癌組織的免疫組織化學染色 (IS) 檢查進一步證實了上述結果，大腸癌組織較其癌旁正常組織表達3.1倍JAG2和6.6倍FZD3蛋白。因此， 我們假設JAG2和FZD3在大腸癌的發生中起重要作用。 / 為了檢驗該假設的真偽，我們運用RNA 干擾的方法進行功能缺失研究。通過該方法，大腸癌細胞株中JAG2 信使RNA和蛋白均能夠被下調，但是FZD3蛋白卻沒有顯示降低。為了弄清JAG2基因的功能，我們進行了單層細胞劃痕傷口癒合試驗和Matrigel 侵襲試驗。結果提示，JAG2 基因下調顯著抑制大腸癌細胞遷移和侵襲的能力。 / 為了調查參與上述功能的機制，我們利用腫瘤轉移相關基因的qPCR陣列分析，試圖檢測出JAG2基因敲除後上調或下調表達的轉移相關基因。結果顯示組織蛋白酶K (CTSK)，一種溶酶體半胱氨酸蛋白酶，在JAG2基因沉默的大腸癌細胞株中表達下調。為了闡明CTSK 活性在大腸癌細胞株侵襲能力中起到的作用，我們採用CTSK抑制劑處理大腸癌細胞株HCT116和DLD-1，發現這兩種細胞株的侵襲能力分別下降了36%和59%。總之, 這些發現表明CTSK可能是JAG2的下游靶基因，活性CTSK可能參與了JAG2介導的大腸癌細胞株侵襲能力。 / 以前的研究表明p38 MAPK通路參與癌細胞遷和侵襲能力的調控。通過Western blot方法，磷酸化的p38和磷酸化的STAT3被發現在JAG2基因沉默的大腸癌細胞中表達降低。p38抑制劑處理的 HCT116和DLD-1細胞降低了侵襲能力下降，同時遷移能力也由於p38抑制劑的處理而降低，支持p38可調控癌細胞遷移和侵襲能力的事實。 / 總之，我們的結果顯示JAG2高表達通過啟動CTSK和p38 MAPK通路，可能促進大腸癌轉移。因此，JAG2可能成為轉移性大腸癌治療的潛在標靶。 / Colorectal cancer (CRC) is one of the most frequent cancers worldwide and is a leading cause of cancer mortality. Around 50% of patients with CRC will experience metastases. Although significant progress has been made in CRC treatment within the last decade with the approval of multiple new chemotherapeutic agents, the prognosis for patients with metastatic CRC remains poor. With the advancement of molecular techniques, novel therapeutic targets are able to be discovered in CRC cell lines and validated in patient samples. / Therefore in this project, I aim to identify potential novel therapeutic targets by comparing the gene expression profile of colon cancer cell lines and a normal colon cell line using quantitative polymerase chain reaction (qPCR) arrays. Results showed that Jagged 2 (JAG2) and Frizzled-3 (FZD3) were up-regulated in the CRC cell lines SW480 and SW620 as compared to the normal colon cell line CCD-18Co. Those results were further validated by immunohistochemical staining (IS), which detected up-regulated JAG2 (3.1-fold) and FZD3 (6.6-fold) proteins expression in CRC tissues as compared to adjacent normal tissues. Thus I hypothesized that JAG2 and FZD3 may play an important role in CRC carcinogenesis. / In order to study the roles of FZD3 and JAG2 in CRC, loss-of-function studies by RNA interference (RNAi) were carried out. While the expression of FZD3 protein failed to be down-regulated by RNAi, JAG2 expression was successfully knocked down in CRC cell lines at both the mRNA and protein levels. Functional analyses using the monolayer scratch wound-healing assay and Matrigel invasion assay showed that JAG2 knockdown significantly inhibited migration and invasion in CRC cell lines. / To investigate the mechanisms involved, a tumour metastasis qPCR array was used to examine the changes in the expression level of metastasis-related genes after JAG2 gene knockdown. Results showed that the expression of Cathepsin K (CTSK), a lysosomal cystein protease, was found to be down-regulated in CRC cell lines following JAG2 silencing. To demonstrate the importance of CTSK activity in CRC cell invasion, HCT116 and DLD-1 CRC cell lines were treated with a CTSK inhibitor and its effect were assessed by the Matrigel invasion assay. Results showed that CTSK inhibition led to a 36% and 59% reduction in number of invaded cells in HCT116 and DLD-1 cell lines, respectively. Taken together, these findings show that CTSK may be a downstream target of JAG2 and that active CTSK may involve in JAG2 mediated invasion in CRC cell lines. / Previous works by others have shown that the p38 MAPK pathway is involved in the regulation of migration and invasive activity of cancer cell lines. Using Western blot analysis, the expression of phosphorylated p38 MAPK and phosphorylated STAT3 were found to be down-regulated following JAG2 depletion in CRC cell lines. In support of a role for p38 MAPK in the regulation of cancer cell migration and invasive capability, treatment with a p38 MAPK inhibitor was found to reduce the percentage of invasive cells and distance moved by migratory cells in HCT116 and DLD-1 cell lines. / In conclusion, my results show that JAG2 over-expression in CRC may promote cancer cell migration and invasion through activation of CTSK and the p38 MAPK pathway. Therefore, JAG2 may be a potential therapeutic target for treatment of metastatic CRC. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / He, Wan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 164-207). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese. / Abstract in English --- p.i / Abstract in Chinese --- p.iv / Acknowledgements --- p.vi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Colorectal Cancer (CRC) --- p.1 / Chapter 1.1.1 --- Epidemiology and Incidence --- p.1 / Chapter 1.1.2 --- Histology --- p.2 / Chapter 1.1.3 --- Gender and Age --- p.4 / Chapter 1.1.4 --- Etiology of CRC --- p.4 / Chapter 1.1.4.1 --- Environment --- p.4 / Chapter 1.1.4.2 --- Hereditary Factors --- p.5 / Chapter 1.1.4.3 --- Dietary Factors --- p.6 / Chapter 1.1.4.4 --- Obesity --- p.6 / Chapter 1.1.4.5 --- Tobacco and alcoho --- p.7 / Chapter 1.1.4.6 --- Inflammatory bowel disease (IBC) --- p.7 / Chapter 1.1.5 --- Genetic Changes in CRC --- p.8 / Chapter 1.1.5.1 --- Chromosomal Aberration --- p.8 / Chapter 1.1.5.2 --- Tumor Suppressor Genes --- p.10 / Chapter 1.1.5.2.1 --- APC gene --- p.10 / Chapter 1.1.5.2.2 --- P53 gene --- p.11 / Chapter 1.1.5.2.3 --- SMAD4 gene --- p.11 / Chapter 1.1.5.3 --- Oncogenes --- p.12 / Chapter 1.1.5.3.1 --- Epidermal Growth Factor Receptor (EGFR) gene --- p.12 / Chapter 1.1.5.3.2 --- RAS gene and BRAF gene --- p.13 / Chapter 1.1.5.4 --- Proposed Two-hit Model for the Multistep Pathogenesis of CRC --- p.15 / Chapter 1.1.6 --- Clinical Presentation and Diagnosis --- p.16 / Chapter 1.1.7 --- Theatment --- p.16 / Chapter 1.1.7.1 --- Surgery --- p.16 / Chapter 1.1.7.2 --- Radiotherapy (RT) --- p.17 / Chapter 1.1.7.3 --- Concurrent Chemotherapy --- p.17 / Chapter 1.1.7.4 --- Target Therapy --- p.18 / Chapter 1.1.7.5 --- Colorectal Cancer Treatment by Stage --- p.19 / Chapter 1.1.7.6 --- Novel Strategies --- p.20 / Chapter 1.1.7.6.1 --- Epigenetic therapy --- p.20 / Chapter 1.1.7.6.2 --- Immunotherapy --- p.21 / Chapter 1.2 --- Pathways Involved in CRC Carcinogenesisand Progression --- p.22 / Chapter 1.2.1 --- Wnt Signaling Pathway --- p.22 / Chapter 1.2.2 --- Notch Signaling --- p.23 / Chapter 1.2.3 --- Nuclear Factor-kappa B (NF-κB) Signaling Pathway --- p.23 / Chapter 1.2.4 --- Phosphatidylinositol 3-kinase (PI3K) Signaling Pathway --- p.24 / Chapter 1.2.5 --- Crosstalk Among WNT, NOTCH, NF-κB and PI3K Signaling Pathway in CRC --- p.24 / Chapter 1.3 --- Hypothesis and Objectives of this Study --- p.28 / Chapter Chapter 2 --- Identification of Differentially Expressed Genes between Colorectal Cancer Cell Lines and A Normal Colon Cell Line --- p.29 / Chapter 2.1 --- Background --- p.29 / Chapter 2.2 --- Materials and Methods --- p.33 / Chapter 2.2.1 --- Cell Lines --- p.33 / Chapter 2.2.2 --- Identification of Differetially Expressed Genes by qPCR Arrays --- p.33 / Chapter 2.2.2.1 --- Total RNA Extraction --- p.33 / Chapter 2.2.2.2 --- RNA Quality Contol --- p.34 / Chapter 2.2.2.3 --- Reverse Transcription (RT) --- p.34 / Chapter 2.2.2.4 --- PCR Arrays --- p.34 / Chapter 2.3 --- Results --- p.36 / Chapter 2.3.1 --- Differentially Expressed Genes in WNT Signaling Pathway --- p.36 / Chapter 2.3.2 --- Differentially Expressed Genes in Notch Signaling Pathway --- p.40 / Chapter 2.3.3 --- Differentially Expressed Genes in NF-κB Signaling Pathway --- p.43 / Chapter 2.3.4 --- Differentially Expressed Genes in PI3K-AKT Signaling Pathway --- p.46 / Chapter 2.3.5 --- Choice of over-expressed genes for further validation and characterization --- p.49 / Chapter 2.4 --- Discussions --- p.53 / Chapter 2.4.1 --- WNT Signaling Pathway --- p.53 / Chapter 2.4.2 --- NOTCH Signaling Pathway --- p.54 / Chapter 2.4.3 --- NF-κB Signaling Pathway --- p.55 / Chapter 2.4.4 --- PI3K-AKT Signaling Pathway --- p.56 / Chapter 2.4.5 --- Choice of over-expressed genes for further validation and characterization --- p.56 / Chapter Chapter 3 --- JAG2, FZD3 and NOTCH4 Expression in Colorectal Cancer Cell Lines and Colorectal Cancer Tissues --- p.59 / Chapter 3.1 --- Background --- p.59 / Chapter 3.1.1 --- JAG2 Ligand --- p.59 / Chapter 3.1.2 --- FZD3 Receptor --- p.61 / Chapter 3.1.3 --- NOTCH4 Receptor --- p.62 / Chapter 3.2 --- Materials and Methods --- p.64 / Chapter 3.2.1 --- CRC Cell Lines --- p.65 / Chapter 3.2.2 --- CRC Tissues --- p.65 / Chapter 3.2.3 --- Quantitative RT-PCR --- p.66 / Chapter 3.2.4 --- Detection of JAG2, FZD3 and NOTCH4 Protein Expression in CRC Tissues by Immunohistochemical Staining (IS) --- p.67 / Chapter 3.2.5 --- Western Blot Assays --- p.68 / Chapter 3.2.5.1 --- Protein extraction --- p.68 / Chapter 3.2.5.2 --- SDS-PAGE gel electrophroresis --- p.68 / Chapter 3.2.5.3 --- Protein blotting --- p.68 / Chapter 3.2.6 --- Detection of JAG2 and FZD3 Protein Expression in CRC and Normal Colon Cell Lines by Western Blotting --- p.69 / Chapter 3.2.7 --- Statistical Analysis --- p.70 / Chapter 3.3 --- Results --- p.71 / Chapter 3.3.1 --- JAG2 and FZD3 but not NOTCH4 mRNA were Over -expressed in CRC Cell Lines --- p.71 / Chapter 3.3.2 --- Over-expression of JAG2 and FZD3 Proteins in CRC Tissues --- p.72 / Chapter 3.3.3 --- FZD3 Over-expression Correlated with Tumour-Node Metastasis (TNM) stages --- p.76 / Chapter 3.3.4 --- JAG2 and FZD3 Protein Expression in Colorectal Cancer and Normal Cell Lines --- p.77 / Chapter 3.4 --- Discussions --- p.78 / Chapter Chapter 4 --- Functional Analyses of JAG2 and FZD3 in CRC Cell Lines by RNA Interference --- p.81 / Chapter 4.1 --- Background --- p.81 / Chapter 4.2 --- Materials and Methods --- p.84 / Chapter 4.2.1 --- Transfection of siRNA into CRC Cell Lines --- p.84 / Chapter 4.2.2 --- Cell Proliferation Assay --- p.85 / Chapter 4.2.3 --- Monolayer Scratch Wound Healing Assay --- p.85 / Chapter 4.2.4 --- Matrigel Invasion Assay --- p.86 / Chapter 4.2.5 --- Statistical Analysis --- p.87 / Chapter 4.3 --- Results --- p.88 / Chapter 4.3.1 --- Knockdown of JAG2 and FZD3 Expression by RNA Interference --- p.88 / Chapter 4.3.2 --- Effect of JAG2 Knockdown on Migration of CRC Cell Lines --- p.91 / Chapter 4.3.3 --- JAG2 Knockdown by siRNA 2 Transfection Reduced Migratory Capability of HCT116, DLD-1and HT29 cell lines --- p.94 / Chapter 4.3.4 --- JAG2 Knockdown Impaired the Invasiveness of HCT116 and DLD-1 Cell Lines --- p.97 / Chapter 4.3.5 --- Decreased Migratory and Invasive Capabilities Induced by JAG2 Knockdown was not Due to Reduced Cell Proliferation --- p.100 / Chapter 4.4 --- Discussions --- p.102 / Chapter Chapter 5 --- NOTCH Pathway Inactivation by JAG2 Silencing Reduces Oncogenic Properties of HT29 but not HCT116 andDLD-1 CRC Cell Lines --- p.106 / Chapter 5.1 --- Background --- p.106 / Chapter 5.2 --- Materials and Methods --- p.109 / Chapter 5.2.1 --- CRC Cell lines --- p.109 / Chapter 5.2.2 --- Pharmacological Inhibition of NOTCH signaling by DAPT --- p.109 / Chapter 5.2.3 --- Combination of DAPT Treatment and JAG2 Silencing by siRNA --- p.109 / Chapter 5.2.4 --- Western Blotting --- p.109 / Chapter 5.2.5 --- Cell Proliferation Assay (MTS Assay) --- p.110 / Chapter 5.2.6 --- Monolayer Scratch Wound Healing Assay --- p.110 / Chapter 5.2.7 --- Matrigel Invasion Assay --- p.111 / Chapter 5.2.8 --- Statistical Analysis --- p.111 / Chapter 5.3 --- Results --- p.112 / Chapter 5.3.1 --- JAG2 Silencing Down-regulates Notch Pathway Signaling in CRC Cell Lines --- p.112 / Chapter 5.3.2 --- Inhibition of NOTCH Signaling by DAPT Treatment in CRC Cell Lines --- p.112 / Chapter 5.3.3 --- NOTCH Inhibition Does not Significantly Affect Cell Proliferation in CRC Cell Lines --- p.114 / Chapter 5.3.4 --- Suppression of NOTCH Signaling by DAPT Inhibits Migration in HT29 but not in HCT116 and DLD-1 CRC Cell Lines --- p.115 / Chapter 5.3.5 --- Suppression of NOTCH Signaling by DAPT does not Significantly Affect Invasiveness of HCT116 and DLD-1 CRC Cell Lines --- p.117 / Chapter 5.4 --- Discussions --- p.118 / Chapter Chapter 6 --- JAG2 Knockdown Inhibits Invasion in CRC Cell Lines through Inactivation of Cathepsin K --- p.121 / Chapter 6.1 --- Background --- p.121 / Chapter 6.2 --- Materials and Methods --- p.123 / Chapter 6.2.1 --- Human Tumour Metastasis RT2 Profiler[superscript TM] PCR Array --- p.123 / Chapter 6.2.2 --- Measurement of CTSK Gene expression level by Quantitative Real-Time PCR --- p.123 / Chapter 6.2.3 --- Immunohistochemical Staining (IS) of CTSK in CRC Tissues --- p.124 / Chapter 6.2.4 --- Pharmacological Inhibitior of CTSK in CRC Cell Lines --- p.124 / Chapter 6.2.5 --- Inhibition of CTSK in CRC Cell Lines for Migration Study --- p.124 / Chapter 6.2.6 --- Inhibition of CTSK in CRC Cell Lines for Invasion Study --- p.125 / Chapter 6.2.7 --- Western Blotting --- p.125 / Chapter 6.2.8 --- Statistical Analysis --- p.125 / Chapter 6.3 --- Results --- p.126 / Chapter 6.3.1 --- Identification of Metastasis Related Genes Which were Down-regulated by JAG2 Knockdown in HCT116 Cells --- p.126 / Chapter 6.3.2 --- Validation of Down-regulation of CTSK Gene by JAG2 Knockdown in HCT116 Cell Line by qRT-PCR --- p.126 / Chapter 6.3.3 --- JAG2 Knockdown Reduced Expression of Active CTSK Protein in CRC Cell Lines --- p.128 / Chapter 6.3.4 --- CTSK Protein Expression in CRC Tissue Samples --- p.130 / Chapter 6.3.5 --- Pharmacological Inhibition of CTSK Suppressed Invasiveness of CRC Cell Lines --- p.131 / Chapter 6.3.6 --- Pharmacological Inhibition of CTSK did not Affect Migration of CRC Cell Lines --- p.132 / Chapter 6.4 --- Discussions --- p.133 / Chapter Chapter 7 --- Depletion of JAG2 Inhibits Migration and Invasion in CRC Cell Lines through Inactivation of p38 MAPK/HSP27 Pathway --- p.137 / Chapter 7.1 --- Background --- p.137 / Chapter 7.2 --- Materials and Methods --- p.140 / Chapter 7.2.1 --- Pharmocological Inhibition of p38 MAPK Phosphorylation CRC Cell Lines --- p.140 / Chapter 7.2.2 --- Inhibition of p38 MAPK Phosphorylation for Migration Study in CRC Cell Lines --- p.140 / Chapter 7.2.3 --- Inhibition of p38 MAPK Phosphorylation for Invasion Study in CRC Cell Lines --- p.140 / Chapter 7.2.4 --- Knockdown of STAT3 by RNA interference --- p.141 / Chapter 7.2.5 --- Knockdown of STAT3 for Migration Study in CRC Cell Lines --- p.141 / Chapter 7.2.6 --- Knockdown of STAT3 for Invasion Study in CRC Cell Lines --- p.141 / Chapter 7.2.7 --- Western Blotting --- p.141 / Chapter 7.2.8 --- Statistical Analysis --- p.142 / Chapter 7.3 --- Results --- p.143 / Chapter 7.3.1 --- JAG2 Knockdown Inhibits p38 MAPK / HSP27 Pathway in CRC Cell Lines --- p.143 / Chapter 7.3.2 --- Inhibition of p38 MAPK / HSP27 Signaling Pathway Down-regulated Invasive Capability of CRC Cell Line --- p.145 / Chapter 7.3.3 --- Inhibition of p38 MAPK / HSP27 Signaling Pathway Down-regulated Migration of CRC Cell lines --- p.147 / Chapter 7.3.4 --- JAG2 Knockdown Inactivated p38 MAPK / HSP27 Pathway Independently of NOTCH Pathway in CRC Cell Lines --- p.149 / Chapter 7.3.5 --- JAG2 Knockdown Inhibits STAT3 Activation in CRC Cell Lines --- p.151 / Chapter 7.3.6 --- STAT3 Silencing Reduced Invasive Capability in CRC Cell Lines --- p.152 / Chapter 7.3.7 --- STAT3 Silencing Reduced Migratory Capability in CRC Cell Lines --- p.154 / Chapter 7.3.8 --- Inhibition of p38 MAPK Activity Suppressed STAT3 Activation in HCT116 Cells --- p.156 / Chapter 7.4 --- Discussions --- p.157 / Chapter Chapter 8 --- Conclusions and Future Works --- p.161 / Chapter 8.1 --- Conclusions --- p.161 / Chapter 8.2 --- Future work --- p.163 / References --- p.164 / Chapter Appendix 1 --- List of Figures and Tables --- p.208 / Chapter Appendix 2 --- Abbrevations used in this thesis --- p.212

Submitted by Suzana Dias (suzana.dias@famerp.br) on 2018-10-29T17:01:35Z No. of bitstreams: 1 CarlaMariaMoraes_dissert.pdf: 4345886 bytes, checksum: 7c2b7d3028309208358698339e534111 (MD5) / Made available in DSpace on 2018-10-29T17:01:35Z (GMT). No. of bitstreams: 1 CarlaMariaMoraes_dissert.pdf: 4345886 bytes, checksum: 7c2b7d3028309208358698339e534111 (MD5) Previous issue date: 2017-07-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Colorectal cancer (CRC) is a prevalent disease in Brazil and worldwide. It is treated with surgical indication, neoadjuvant therapies such as Chemotherapy and / or Radiotherapy. Thus in elective surgeries, the approach of the multidisciplinary team to improve the clinical condition of surgical patients is required. Nutritional impairments to the side effects of cancer treatment prior to surgery are permanent. Therefore, the preoperative nutritional assessment, therapy and post-surgical nutritional monitoring are fundamental, since this nutritional impairment contributes to the increase of postoperative morbidity and mortality. Objective: To identify the status and preoperative nutritional risk of patients eligible for colorectal surgery. Methods: We studied from February, 2014 to August, 2015, 22 patients (11 men and 11 women; 59.6± 12.4 years) hospitalized in the Coloproctology Department of Hospital de Base/ FAMERP candidates to the elective major abdominal surgery. Preoperative evaluation was performed by a protocol that comprised subjective global assessment (SGA), objective nutritional assessment (ONA), anthropometric parameters (weight, height, BMI, measures of circumferences and triceps skinfold), biochemical (hematocrit, hemoglobin and serum albumin) and dietary recall of the usual intake. Nutritional risk was calculated according to criteria combined according to the Nutritional Guidelines Nutritional Therapy (DITEN) and the Second National Consensus Oncology Nutrition (CNNO) based on parameters such as: weight loss (pp)>10% in 6 months, severe malnutrition according to SGA, BMI <18.5kg/m2 (adults) or 22 kg/m2 (elderly) and serum albumin <3.0g/dL. Results: In the study, 18 (81.8%) patients had cancer (50.0% colon cancer and 31.8% rectal cancer). All patients had major surgery indication, 8 (27.3%) reported having gastrointestinal symptoms, 20 (90.9%) reported no impairment in functional capacity and 8 (36.4%) showed depletion of adipose tissue and lean mass. Nine (40.9%) patients were considered malnourished or at nutritional risk according to SGA, and only 2 (9.1%) according to BMI (P=0.0339). Thirteen (59.1%) of them were overweighed according to BMI, 7 (31.8%) and 11 (50%) were classified with malnutrition / nutritional risk and overweight according to ONA, respectively. Regarding the anthropometric profile, the group of men towards women; higher weight median (77.8kg x 60.0 kg, P=0.0126), CMB (26.4 cm x 22.2 cm P=0.0372), DC (100 cm x 80 cm; P=0.0301) and WHR (0.99 x 0.88, P=0.0010) were observed. Regarding the PCT, there was a higher median in the female group compared to the group of men (23.0 mm x 10.0 mm; P=0.0065). In 10 (45.5%) patients and in 11 (50%) of them, the ingestion of calories and protein was smaller than 75% of the required. Only 2 (9.1%) patients had serum albumin levels between 3.5 and 3.0 g/dL. Thus, 4 (18.2%) patients were considered to have severe nutritional risk according to DITEN and II CNNO. Conclusions: Considering the results found in the study, it is concluded that the impaired nutritional status, either by depletion of lean body mass, as the overweight is a common finding in patients undergoing surgery for colorectal cancer. Severe nutritional risk is present in these patients, so the Preoperative Nutritional Therapy is fundamental and can provide better results in the elective surgeries. / O câncer colorretal (CCR), doença prevalente no Brasil e no mundo, a qual é tratada com cirúrgia, terapias neoadjuvantes e adjuvantes, como quimioterapia e/ou radioterapia vem ganhando mais atenção devido ao aumento da incidência. Assim, em cirurgias eletivas, a atuação da equipe multidisciplinar pode contribuir decisivamente para melhorar as condições clínicas dos pacientes cirúrgicos e o resultado do tratamento. Prejuízos nutricionais devido aos efeitos colaterais do tratamento oncológico anterior à cirurgia são evidentes. Portanto, a avaliação nutricional pré-cirúrgica, a terapia e o acompanhamento nutricional pós-cirúrgico são fundamentais, uma vez que o comprometimento nutricional contribui para o aumento da morbimortalidade pós-operatória. Objetivo: Identificar o estado e o risco nutricional pré-operatório de pacientes candidatos à cirurgia colorretal. Casuística e Método: Foram estudados, de Fevereiro/2014 à Agosto∕2015, 22 pacientes (11 homens e 11 mulheres; 59,6+ 12,4 anos), internados nas enfermarias da Disciplina de Coloproctologia do Hospital de Base/FAMERP, candidatos à cirurgia abdominal eletiva de grande porte. A avaliação pré-operatória foi realizada por protocolo composto de avaliação nutricional subjetiva global (ANSG), avaliação nutricional objetiva (ANO), parâmetros antropométricos (peso, altura, IMC, medidas de circunferências e prega cutânea do tríceps), bioquímicos (hematócrito, hemoglobina e albumina sérica) e recordatório alimentar da ingestão habitual. O risco nutricional foi calculado de acordo com critérios combinados segundo as Diretrizes Nutricionais de Terapia Nutricional (DITEN) e o II Consenso Nacional de Nutrição Oncológica (CNNO), baseado em parâmetros como: perda de peso (pp)>10% em 06 meses, desnutrição grave de acordo com ANSG, IMC<18,5Kg/m2 (adultos) ou 22Kg/m2 (idosos) e albumina sérica <3,0g/dL. Resultados: Na casuística, 18(81,8%) eram portadores de câncer (50,0%câncer de cólon e 31,8% câncer de reto) e 18,2 outras doenças do intestino. Todos os pacientes tinham indicação de cirurgia de grande porte, sendo que 6(27,3%) relataram apresentar sintomas gastrointestinais, 20(90,9%) referiram ausência de disfunção na capacidade funcional e 8(36,4%) apresentaram depleção de tecido adiposo e de massa magra. Nove (40,9%) pacientes foram considerados desnutridos ou de risco nutricional segundo a ANSG, e apenas 2(9,1%) segundo o IMC (P=0,0339). Treze (59,1%) deles tinham excesso de peso de acordo com o IMC, 7(31,8%) e 11(50%) classificaram-se com desnutrição/risco nutricional e com excesso de peso segundo a ANO, respectivamente. Em relação ao perfil antropométrico, no grupo dos homens em relação às mulheres, observaram-se maiores medianas de peso (77,8kg x 60,0kg; P=0,0126), CMB (26,4cm x 22,2cm; P=0,0372), CC (100 cm x 80 cm; P=0,0301) e RCQ (0,99 x 0,88; P=0,0010). Em relação ao PCT, verificou-se maior mediana no grupo das mulheres comparado ao grupo dos homens (23,0mm x 10,0mm; P=0,0065). Em 10(45,5%) pacientes e em 11(50%) deles, a ingestão de calorias e proteínas foi considerada menor que 75% das necessidades. Apenas 2(9,1%) pacientes apresentaram níveis séricos de albumina entre 3,5 a 3,0 g/dL. Sendo assim, 4 (18,2%) pacientes foram considerados com risco nutricional grave segundo o DITEN e o II CNNO. Conclusões: Considerando-se os resultados encontrados na casuística, conclui-se que o estado nutricional comprometido, tanto pela depleção de massa magra, como pelo excesso de peso, é um achado frequente em pacientes que realizam a cirurgia do câncer colorretal. O risco nutricional grave está presente nestes pacientes, de tal modo que a Terapia Nutricional pré-operatória, é fundamental, pois pode influenciar os melhores resultados em cirurgias eletivas.

O câncer colorretal (CCR) é a neoplasia mais prevalente no mundo e, cerca de 60% apresentarão metástases hepáticas, representando uma importante causa de mortalidade. Aproximadamente 35% dos pacientes apresentam metástases hepáticas no momento do diagnóstico do tumor primário ou desenvolverão metástases durante o primeiro ano após o tratamento da neoplasia colorretal, conhecidas como metástases sincrônicas. Inúmeros trabalhos têm demonstrado que as metástases sincrônicas representam importante fator prognóstico negativo na evolução destes pacientes, representando um grupo de pacientes que necessita uma abordagem multidisciplinar mais agressiva. O objetivo deste trabalho foi de avaliar os resultados do tratamento cirúrgico das metástases sincrônicas de CCR e determinar os possíveis fatores que pudessem interferir no prognóstico de sobrevida livre de doença e sobrevida atuarial. Entre maio de 1996 e dezembro de 2007, 59 pacientes submetidos à ressecção hepática por metástases sincrônicas foram avaliados retrospectivamente através de análise uni e multivariada. A mortalidade pós-operatória foi de 3,38%, e a morbidade pós-operatória de 30,50%. A sobrevida estimada em 5 anos foi de 38,45% e a sobrevida livre de doença no mesmo período foi de 23,96. O valor do antígeno carcinoembrionário igual ou superior a 50 ng/ml e o número de metástases hepáticas maior que três lesões representaram fatores prognósticos limitados da sobrevida livre de doença, porém sem interferir na sobrevida atual. Pacientes com metástases no fígado e com doença extra-hepática, selecionados para a ressecção, não apresentaram sobrevida livre de doença acima de 20 meses, porém sem impacto na sobrevida a longo prazo. Nenhum dos fatores prognósticos estudados interferiu na sobrevida atual tardia. Entretanto,não foi observada sobrevivência além de 40 meses em pacientes com mais de três metástases hepáticas. A ressecção de metástases sincrônicas de câncer colorretal pode propiciar sobrevida tardia em mais de um terço dos pacientes. O valor do CEA e do número de metástases representaram fatores prognósticos limitantes da sobrevida livre de doença / Colorectal cancer is the world´s most prevalent digestive neoplasia and about 60% of the patients will present liver metastases, representing an important cause of mortality. About 35% of the patients present hepatic metastases at the diagnosis of the colorectal tumor or will develop metastases during the first year after the treatment of the primary tumor, known as synchronous metastases. Innumerable studies have shown that synchronous metastases represent a negative prognostic factor in the evolution of these patients, representing a group of patients that need an aggressive multidisciplinary approach. The purpose of this study was to evaluate results of the surgical treatment of colorectal cancer synchronous metastases and to determine possible factors that might interfere in the prognosis of disease-free and actuarial survival. Between May 1996 and December 2007, 59 patients submitted to liver resection for synchronous metastases were retrospectively evaluated through univariate and multivariate analysis. Postoperative morbidity and mortality were 30.5% and 3.38%, respectively. Cumulative survival estimated in 5 years was 38.45% and disease-free survival in the same period was 23.96%. Levels of carcino-embrionary antigen (CEA) higher than 50 ng/ml and the number of hepatic metastases higher than three lesions represented negative prognostic factors limiting disease-free survival; however, with no impact on cumulative survival. Patients with liver metastases and extrahepatic disease selected for resection didnt present a disease-free survival above 20 months, yet without impact in global survival. None of the prognostic factors studied interfered in long term actuarial survival, however survival beyond 40 months in patients with more than three hepatic metastases was not observed. Resection of synchronous metastases of colorectal cancer may provide late survival in more than one third of the patients. CEA values and the number of metastases represented prognostic factors with negative impact on disease-free survival

Introdução: Atualmente a ressecção hepática é o melhor tratamento para metástases de câncer colorretal. Diversos fatores prognósticos foram estudados e muitos estudos têm demonstrado que metástases nos linfonodos hilares constituem um fator prognóstico adverso. Este estudo avaliou a frequência e as características do envolvimento linfonodal hilar microscópico, através de linfadenectomia sistemática, com pesquisa de micrometástases em pacientes submetidos à hepatectomia por metástases colorretais. Métodos: Os critérios de exclusão foram: irressecabilidade detectada no pré e intraoperatório, condições clínicas e fatores intraoperatórios que exigiam menor tempo cirúrgico, metástases linfonodais macroscópicas confirmadas por exame de congelação e menos de três linfonodos no produto da linfadenectomia. De 38 pacientes iniciais, 28 foram submetidos ressecção hepática em associação com linfadenectomia sistemática do hilo, apresentando três ou mais linfonodos dissecados. Os linfonodos negativos ao método convencional de hematoxilina e eosina foram avaliados através de cortes seriados com intervalos de 100 m associada à imunoistoquímica com anticorpos contra pancitoqueratinas humanas AE1/AE3. Resultados: Em média, 6,18 linfonodos foram dissecados por paciente. A linfadenectomia aumentou o tempo operatório em 48 minutos, no entanto, não houve morbimortalidade associada a este procedimento. Dois pacientes (7,2%) apresentaram metástase linfonodal microscópica ao exame convencional com hematoxilina e eosina. Quando aplicado os cortes seriados associados à imunoistoquímica, três pacientes adicionais (10,8%) foram identificados como portadores de micrometástases linfonodais. Conclusão: A frequência global de metástases microscópicas, incluindo micrometástases foi de 18%. Não houve correlação estatística entre outros fatores prognósticos e a presença de metástases microscópicas. A linfadenectomia sistemática associada à pesquisa de micrometástases ampliou a detecção de envolvimento linfonodal microscópico, contribuindo assim, com o estadiamento de doença extra-hepática / Introduction: Currently, hepatectomy is considered the best treatment of metastatic colorectal cancer. Several prognostic factors have been investigated, and many studies have shown that the involvement of regional lymph nodes at the hepatic hilum represents a negative prognostic factor. The present study investigated the frequency and characteristics of microscopic involvement of hilar lymph nodes, through systematic lymphadenectomy and analysis of micrometastases in patients undergoing hepatectomy due to colorectal metastases. Methods: Exclusion criteria were: no resectable disease detect in the pre or intra-operative; clinics conditions and intraoperative factors what required minor surgical time; macroscopic hepatic lymph node metastases, confirmed by frozen section and less of three lymph nodes resected for patient. Of the 38 patients, 28 underwent hepatic resection in association with systematic lymphadenectomy of the hepatic hilum, with three or more lymph nodes resected for patient. Lymph nodes considered negative by conventional hematoxylin and eosin staining were analyzed by serial sectioning with 100 m intervals and immunohistochemistry with antibodies to cytokeratins AE1/AE3. Results: In average, 6.18 lymph nodes were dissected per patient. Lymphadenectomy increased surgical time by 48 minutes in average, but no morbi-mortality was associated to the procedure. In two of the patients (7,2%), conventional hematoxylin and eosin analysis showed the presence of microscopic lymph node metastases. Immunohistochemistry analysis of serial sections allowed the identification of three other patients with lymph node micrometastases (10,8%). Conclusion: The overall frequency of microscopic metastases, including micrometastases, was 18%. No statistically significant relationships were observed between other prognostic factors and the presence of microscopic metastases. Systematic lymphadenectomy with inclusion of micrometastases protocols improved the detection of microscopic lymph node involvement, resulting in more accurate staging of extrahepatic disease

Background & Aims: By whole genome sequencing, we identified for the first time that solute carrier family 12 member 5 (SLC12A5) gene located on chromosome 20q13.12 was amplified in colorectal cancer (CRC). We aimed to determine the amplification status of SLC12A5 and its clinical implication in CRC, and characterize the functional mechanisms of SLC12A5 in colorectal carcinogenesis. / Materials and Methods: Protein expression level of SLC12A5 was evaluated by immunohistochemistry. SLC12A5 amplification was verified by fluorescence in situ hybridization (FISH). The correlations between SLC12A5 expression and clinicopathologic parameters as well as the prognosis impact of SLC12A5 were analyzed in 195 CRC patients. The biological function of SLC12A5 in CRC cell lines were determined by cell viability, colony formation, invasion, migration, flow cytometry and in vivo tumorigenicity assays. Standard tail vein metastatic assay was performed to examine the effect of SLC12A5 in lung metastasis in nude mice. Western blot, luciferase reporter assays and human tumor metastasis PCR array were performed to evaluate SLC12A5 downstream effectors and related pathways. / Results: RT-PCR showed SLC12A5 was readily expressed in 7 of 9 CRC cell lines, but was absent in normal colorectal tissues. The mean protein expression level of SLC12A5 was significantly higher in primary CRCs as compared to their adjacent normal tissues. Amplification of SLC12A5 was detected in 40.8% (78/191) of primary CRCs by FISH, which was positively correlated with its protein overexpression (P < 0.001). Overexpression of SLC12A5 was positively associated with a more advanced TNM stage (P < 0.05). Multivariate Cox regression analysis showed that SLC12A5 overexpression was an independent predictor of poorer survival of CRC patients (P = 0.018). We further tested the biological function of SLC12A5 in human colon cancer cells. Ectopic expression of SLC12A5 in colon cancer cells SW480 and SW1116 increased proliferation and colony formation. Silencing SLC12A5 expression in HCT116 by siRNA had the opposite effects in vitro, and knockdown of SLC12A5 by shRNA significantly inhibited xenograft tumor growth in nude mice. We further revealed that SLC12A5 inhibited apoptosis of colon cancer cells by mediating apoptosis-inducing factor (AIF) and endonuclease G (EndoG) -dependent apoptotic signaling pathway. Moreover, gain-and loss-of-function experiments showed that SLC12A5 enhanced cell invasion and migration in vitro. Knockdown of SLC12A5 by shRNA significantly inhibited lung metastasis in nude mice. SLC12A5 promoted tumor metastasis through regulating key elements of the matrix architecture, such as matrix metallopeptidase and fibronectin. / Conclusion: We have identified a novel amplification gene SLC12A5 which is overexpressed in CRC. SLC12A5 may be an independent prognostic marker for CRC and may play a pivotal oncogenic role in colorectal carcinogenesis by inhibiting apoptosis and promoting metastasis. / 背景和目的：通過對結直腸癌進行全基因組測序，我們首次發現位於染色體20q13.12的SLC12A5基因在結直腸中擴增。本研究旨在探索SLC12A5在結直腸癌中的擴增情況和臨床意義，并進一步研究SLC12A5在結直腸癌發生發展中的作用機制。 / 材料和方法：採用免疫组化方法檢測SLC12A5的蛋白表达水平。應用熒光原位雜交方法驗證SLC12A5基因的擴增情況。在195例結直腸癌患者中对SLC12A5表达與临床病理關係及其對預後的影響其进行分析。通过檢測細胞活力、細胞集落形成實驗、侵襲實驗、遷移實驗、流式細胞術和體內成瘤實驗以研究SLC12A5在結直腸癌中的生物学功能。進而通過免疫印跡、熒光素酶報告實驗和人腫瘤轉移的PCR陣列，探索SLC12A5調控的基因和相关途径。 / 结果：我們採用RT-PCR方法檢測SLC12A5在9株結直腸癌細胞株的表達情況，SLC12A5在7株結直腸癌細胞株中穩定表達，但是在正常大腸組織中表達沉默。SLC12A5在結直腸中的平均蛋白表達水平顯著高於其鄰近的正常組織。通過熒光原位雜交方法，在40.8% （78/ 191）的結直腸癌中檢測到SLC12A5的擴增，該基因的擴增與其蛋白高表達水平呈正相關關係。SLC12A5高表達水平跟晚期TNM分期密切相關（P <0.05）。多因素Cox回歸分析表明，SLC12A5高表達是結直腸癌患者較差的生存的獨立預測因子（P = 0.018）。我們進一步在人結腸癌細胞株中檢測SLC12A5的生物功能。在結腸癌細胞SW480和SW1116中過度表達SLC12A5促進細胞增殖和集落形成。siRNA敲低HCT116 細胞SLC12A5的表達在體外實驗中有相反的效果。此外，shRNA敲低SLC12A5的表達顯著抑制裸鼠移植瘤的生長。我們進一步發現，SLC12A5通過介導凋亡誘導因子（AIF）和核酸內切酶G（EndoG）-依賴的細胞凋亡信號轉導通路抑制結腸癌細胞的凋亡。此外，功能獲得性和功能缺失性的體外實驗表明，SLC12A5促進腫瘤細胞的侵襲和遷移。尾靜脈注射實驗表明shRNA敲低SLC12A5的表達顯著抑制裸鼠肺轉移。SLC12A5通過調節基質結構的關鍵因子，如基質金屬蛋白酶和纖維連接蛋白，促進腫瘤轉移。 / 结论：我們發現了一個新的擴增基因SLC12A5，該基因在結直腸癌中高表達。SLC12A5是結直腸癌的一個獨立的預後標誌物。SLC12A5通過抑制細胞凋亡和促進腫瘤轉移，在結直腸癌的發生發展中起了舉足輕重的致癌作用。 / Xu, Lixia. / Thesis Ph.D. Chinese University of Hong Kong 2014. / Includes bibliographical references (leaves 107-120). / Abstracts also in Chinese. / Title from PDF title page (viewed on 05, October, 2016). / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only.

INTRODUCCIÓ L’angiogènesi és essencial per al creixement dels tumors sòlids i participa en el procés metastàtic. El VEGF-C, un potent factor angiogènic, es troba elevat en pacients amb càncer colorectal (CCR). Hi ha evidències que suggereixen que la COX-2 és un factor important en el procés de carcinogènesi colorectal. L’objectiu principal de l’estudi és investigar l’efecte que un inhibidor selectiu de la ciclooxigenasa-2, pugui tenir a un model de carcinogènesi colònica experimental sobre la superfície tumoral del còlon, i la seva traducció als nivells sèrics de VEGF-C i a l’expressió de ciclooxigenasa-2 (COX-2). MATERIAL I MÈTODES Es van utilitzar 100 rates Sprague-Dawley no consanguínies distribuïdes en cinc grups: grup control, grup amb CCR induït amb DMH, grup de rates tractades amb celecoxib, grup de rates tractades amb àcid acetilsalicílic i grup de rates tractades amb indometacina. Després del sacrifici, es van prendre mostres de sang per determinar el VEGF-C sèric mitjançant ELISA i mostres dels segments del còlon i d’aquells suggestius de contenir neoplàsia per a estudiar l’expressió de COX-2 als tumors i a la mucosa normal del còlon descendent mitjançant immunohistoquímica. Es va utilitzar la prova estadística més idònia a cada supòsit d’estudi i es va considerar significatiu quan la p£0,05. RESULTATS Vam trobar expressió positiva de COX-2 en el 57,14-66,67% dels tumors de còlon. El 72% de tumors carcinomes mucinosos van presentar un score de tinció de COX-2 positiu, vs. un 48,46% dels adenocarcinomes (p<0,05). Es van trobar diferències significatives entre els grups que van rebre fàrmacs quan es va comparar el nivell sèric de VEGF-C (p=0,008). El grup que va prendre àcid acetilsalicílic presentà uns nivells sèrics menors als del grup que va rebre celecoxib (63,67 vs. 92,58 pg/ml) (p=0,003). No es van apreciar diferències significatives entre el grup que va rebre celecoxib i el grup control amb inducció tumoral (92,58 vs. 89,89 pg/ml; p=0,698). No es van trobar diferències estadísticament significatives entre els valors de VEGF-C sèric i l’expressió tumoral de COX-2 a cap dels grups d’estudi (p>0,05). Es van trobar diferències significatives entre el valor sèric mig de VEGF-C i l’extensió de l’expressió de COX-2 en el tumor (p=0,035). També es va trobar correlació estadísticament significativa entre el VEGF-C sèric i la suma de superfície tumoral (p=0,045). Es van trobar diferències estadísticament significatives en el VEGF-C sèric segons els animals presentessin o no metàstasis a distància o hematògenes (p=0,015). CONCLUSIONS La major part de tumors de còlon van expressar COX-2. L’expressió de COX-2 es va associar al tipus de tumor, i s’incrementa d’acord amb l’augment de la malignitat de la histologia del tumor. El VEGF-C sèric es relaciona amb la progressió de la malaltia. Valors elevats es van correlacionar amb superfícies tumorals altes de manera estadísticament significativa. Celecoxib no va reduir de manera significativa l’expressió de COX-2 als tumors respecte dels grups tractats amb fàrmacs no selectius, ni va disminuir de manera significativa els valors sèrics de VEGF-C, ni la suma de superfície tumoral en el còlon. L’àcid acetilsalicílic indueix una disminució en els nivells sèrics de VEGF-C, propers als nivells control, però no van existir canvis en l’expressió tumoral de COX-2, ni en la suma de superfície tumoral en comparar aquest grup amb els altres. / INTRODUCCIÓN La angiogénesis es esencial para el crecimiento de los tumores sólidos y participa en el proceso metastático. El VEGF-C, un potente factor angiogénico, se encuentra elevado en pacientes con cáncer colorrectal (CCR).Hay muchas evidencias que sugieren que la COX-2 es un factor importante en el proceso de carcinogénesis colorrectal. El objetivo principal del estudio es investigar el efecto que un inhibidor selectivo de la ciclooxigenasa-2, comparándolo con otros no selectivos, pueda tener, en un modelo de carcinógesis colónica experimental, sobre la superficie tumoral del colon y su traducción en los niveles séricos de VEGF-C y en la expresión de ciclooxigenasa-2 (COX-2). MATERIAL Y MÉTODOS Se utilizaron 100 ratas Sprague-Dawley no consanguíneas distribuidas en cinco grupos: grupo control, grupo con CCR inducido con DMH, grupo de ratas tratadas con celecoxib, grupo de ratas tratadas con ácido acetilsalicílico y grupo de ratas tratadas con indometacina. Tras el sacrificio, se tomaron muestras de sangre para determinar el VEGF-C sérico mediante ELISA y muestras de los segmentos del colon y de aquellos sugestivos de contener neoplasia para estudiar la expresión de COX-2 en los tumores y en la mucosa normal del colon descendente mediante inmunohistoquímica. Se utilizó la prueba estadística más idónea en cada supuesto de estudio y se consideró significativo cuando la p£ 0,05. RESULTADOS Encontramos expresión positiva de COX-2 en el 57,14 - 66,67% de los tumores de colon. El 72% de tumores carcinomas mucinosos presentaron un score de tinción de COX-2 positivo, vs. un 48,46% de los adenocarcinomas (p<0,05). Se encontraron diferencias significativas entre los grupos que recibieron fármacos cuando se comparó el valor sérico de VEGF-C (p=0,008). El grupo que tomó ácido acetilsalicílico presentó unos niveles séricos menores a los del grupo que recibió celecoxib (63,67 vs. 92,58 pg/ml) (p=0,003). No se apreciaron diferencias significativas entre el grupo que recibió celecoxib y el grupo control con inducción tumoral (92,58 vs. 89,89 pg/ml.) (p=0,698). No se encontraron diferencias estadísticamente significativas entre los valores de VEGF-C sérico y la expresión tumoral de COX-2 en ninguno de los grupos de estudio (p>0,05). Se encontraron diferencias significativas entre el valor sérico medio de VEGF-C y la extensión de la expresión de COX-2 en el tumor (p=0,035). También se halló correlación estadísticamente significativa entre el valor circulante de VEGF-C sérico y la suma de superficie tumoral (p=0,045). Se encontraron diferencias estadísticamente significativas en el valor sérico de VEGF-C según los animales presentasen o no metástasis a distancia o hematógenas (p=0,015). CONCLUSIONES La mayoría de tumores de colon inducidos experimentalmente en la rata expresaron COX-2. La expresión de COX-2 en el CCR se asocia de forma significativa al tipo de tumor, incrementándose conforme aumentaba la malignidad de la histología del tumor. El VEGF-C sérico se relaciona con la progresión de la enfermedad. Valores elevados se correlacionaron con sumas de superficies tumorales elevadas de un modo estadísticamente significativo. Celecoxib no redujo de un modo significativo la expresión de COX-2 en los tumores respecto de los grupos tratados con fármacos no selectivos, ni disminuyó de forma significativa los valores séricos de VEGF-C, ni la suma de superficie tumoral en el colon. El ácido acetilsalicílico induce una disminución en los niveles séricos circulantes de VEGF-C cercanos a los niveles control. Sin embargo, no existieron cambios en la expresión de COX-2 en los tumores, ni en la suma de superficie tumoral al comparar este grupo con los demás. / INTRODUCTION Angiogenesis is essential for the growth of solid tumours and takes part in the metastatic process. VEGF-C, a strong angiogenic factor, is increased in patients with colorectal cancer (CRC). There are evidences suggesting that COX-2 is an important factor in the colorectal carcinogenesis process. The main aim of the study is to investigate the effect that a selective COX-2 inhibitor, comparing it with non-selective ones, could have on experimental colon carcinogenesis, on the colorectal tumour area, and its results in the serum VEGF-C levels and in the cyclooxigenase-2 (COX-2) expression. MATERIAL AND METHODS 100 Sprague-Dawley rats without consanguinity were used. They were divided into five groups: control group, group with DMH induced CRC, group that was treated with celecoxib, group that was treated with aspirin and group that was treated with indomethacin. Blood samples were taken after slaughter of rats, in order to determine serum VEGF-C with ELISA technique. Colon segments samples and specimens from the areas that were suggested to contain neoplasms were taken too, in order to study COX-2 expression in colon tumors and in the normal mucosa with immunohistochemistry technique. The most appropriate statistical test was used in each case of the study and it was considered significant when p£ 0.05. RESULTS We found positive COX-2 expression in 57.14-66.67% of colon tumours. 72% of mucinous carcinomas presented a positive score for COX-2 expression, vs. 48.46% of adenocarcinomas (p<0.05). We found significant differences between groups that received drugs when we compared the serum VEGF-C level (p=0.008). The AAS group presented lower serum VEGF-C levels than the celecoxib group (63.67 vs. 92.58 pg/ml) (p=0.003). We didn’t appreciate significant differences between the celecoxib group and the control group with tumor induction (92.58 vs. 89.89 pg/ml) (p=0.698). There were no statistically significant differences between the serum VEGF-C levels and tumor expression of COX-2 in any of the groups of the study (p>0.05). We found significant differences between the serum VEGF-C level and the extension of COX-2 expression in the tumor (p=0.035). We also found statistically significant correlation between the value of serum VEGF-C and the sum of tumor area (p = 0.045). Statistically significant differences were found in the serum VEGF-C level between animals having or not hematogenous metastasis (p=0.015). CONCLUSIONS Most experimentally induced colon tumors in rats expressed COX-2. The expression of COX-2 in CRC is significantly associated with the type of tumor, and increases according to the increases of malignancy of the tumor histology. Serum VEGF-C is related to disease progression. High values were correlated with high sum of tumor surfaces in a statistically significant manner. Celecoxib didn’t reduce significantly the expression of COX-2 in tumors regarding groups treated with non-selective drugs, nor significantly decreased serum VEGF-C, or the sum of tumor area in the colon. Aspirin decreases serum VEGF-C levels which were similar to control levels. However, there were no changes in the expression of COX-2 in tumors, or the sum of tumor area by comparing this group with other treated groups.

Thesis (Ph. D.)--University of Washington, 2003. / Vita. Includes bibliographical references (leaves 117-126).

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 65-82).