>Magister Scientiae - MSc / Prostate Cancer is the leading cause of cancer-related death in males in the Western world. It is a common biological disease originating from the reproductive system of the male namely, the prostate gland, usually in older patients (over the age of 50) and with a family history of this disease. The disease shows clinical aggressiveness due to genetic alterations of gene expression in prostate epithelial cells. Prostate cancer is currently diagnosed by biopsy and prostate cancer screening via the Prostate-Specific Antigen (PSA) blood test. Early detection is critical and although PSA was discovered to aid in the diagnoses of this cancer at its early stages, it has a disadvantage due to its low specificity thus causing unnecessary biopsies of healthy individuals and overtreatment of patients. Although various studies and efforts have been made to identify the ideal biomarker for prostate cancer and many even being applied to clinical use, it is still challenging and has not replaced the best-known biomarker PSA. PSA test has minimal invasive characteristics, at relatively low cost together with high sensitivity but low specificity. Biomarker discovery is a challenging process and a good biomarker has to be sensitive, specific and its test highly standardized and reproducible as well as identify risk for or diagnose a disease, assess disease severity or progression, predict prognosis or guide treatment. Computational biology plays a significant role in the discovery of new biomarkers, the analyses of disease states and the validation of potential biomarkers. Bioinformatic approaches are effective for the detection of potential micro ribonucleic acid (miRNA) in cancer. Altered miRNA expression may serve as a biomarker for cancer diagnosis and treatment. Small non-protein coding RNA, miRNA are small regulatory RNA molecules that modulate the expression of their target genes. miRNAs influence numerous cancer-relevant processes such as proliferation, cell cycle control, apoptosis, differentiation, migration and metabolism. Discovery and existence of extracellular miRNAs that circulate in the blood of cancer patients has raised the possibility that miRNAs may serve as novel diagnostic markers. Since a single miRNA is said to be able to target several mRNAs, aberrant miRNA expression is capable of disrupting the expression of several mRNAs and proteins. Biomarker discovery for prostate cancer of mRNA and miRNA expression are strongly needed to enable more accurate detection of prostate cancer, improve prediction of tumour aggressiveness and facilitate diagnosis. The aim of this project was to focus on functional analyses of genes and their protein products regulated by previously identified miRNA in prostate cancer using bioinformatics as a tool. Most proteins function in collaboration with other proteins and therefore this study further aims to identify these protein-protein interactions and the biological relevance of these interactions as it relates to Prostate cancer. Various computational databases were used such as STRING, DAVID and GeneHub-GEPIS for functional analyses of these miRNA regulated genes. The main focus was on the 21 genes regulated by several miRNAs identified in a previous study. Results from this study identified six genes; ERP44, GP1BA, IFNG, SEPT2, TNFRSF13C and TNFSF4, as possible diagnostic biomarkers for prostate cancer. These results are promising, since the targeted biomarkers would be easily detectable in bodily fluids with the Gene Ontology (GO) analysis of these gene products showing enrichment for cell surface expression. The six genes identified in silico were associated to transcription factors (TFs) to confirm regulatory control of these TFs in cancer promoting processes and more specifically prostate cancer. The CREB, E2F, Nkx3-1 and p53 TFs were discovered to be linked to the genes IFNG, GP1BA, SEPT2 and TNFRSF13C respectively. The expression of these TFs show strong association with cancer and cancer related pathways specifically prostate cancer and thus demonstrates that these genes can be assessed as possible biomarkers for prostate cancer. The prognostic and predictive values of the candidate genes were evaluated to assess their relationship to prognosis of this disease by means of several in silico prognostic databases. The results revealed expression differences for the majority of the candidate genes were not significantly sufficient to be distinguished as strong prognostic biomarkers in several prostate cancer populations. Although one marker, GP1BA was supported as having prognostic value for prostate cancer based on it's statistical pvalue in one of the prostate cancer patient datasets used. Another candidate gene SEPT2 showed promise as it has some prognostic value in the early stages of the disease. Although the results yielded, based on the in silico analysis, were not the discovery of an ideal diagnostic marker based on the set criteria in this study, further analysis using a molecular approach qRT-PCR can be considered for a detailed followup study on selected candidate genes to evaluate their roles in disease initiation and progression of prostate cancer using cell lines as well as patient samples. / CSIR

Numerous studies have demonstrated that tumour cells have the ability to alter immune function to create an immune suppressed environment. This allows tumour cells to escape immune surveillance and consequently the tumour can progress. Dendritic and T cells have critical roles in immune activation and tolerance and are thus major targets of tumour-mediated immune suppression. Understanding the mechanism(s) by which tumour cells modulate the immune system will facilitate the development of immune system-based therapies for cancer treatments. In this study we sought to determine the nature of, and cellular and molecular mechanisms underlying, changes in immune status during tumour progression using mouse models of prostate cancer. Detailed analysis of the immunological status in a mouse prostate dysplasia model (12T-7slow) revealed that immune suppression accompanied tumour progression. We found that T cells isolated from tumour-bearing hosts were hypo-responsive to antigen stimulation. Furthermore, we demonstrated that CD4+CD25+ regulatory T cells were responsible, at least in part, for this alteration. Anti-CD25 antibody treatment reduced, but did not prevent, tumour growth in either a transplanted prostate tumour model or a spontaneously developing prostate tumour model. In addition, an altered dendritic cell phenotype and an elevated frequency of CD4+CD25+ regulatory T cells were observed within the tumour mass. Similar alterations were observed in the prostate-specific Pten knockout mice which develop advanced prostate adenocarcinoma. Interestingly, evidence of immune activation, such as an increased frequency of activated T cells, was detected in the tumour microenvironment in both mouse prostate tumour models. To identify factors that may play critical roles in the altered immune cell phenotype observed in the tumour microenvironment, a global gene expression profiling analysis was carried out to evaluate the changes in immune-related gene expression patterns. This analysis provided additional evidence for the co-existence of immune suppression and immune activation. Moreover, subsequent analyses suggested that one differentially expressed transcript, interferon regulatory factor 7, and its target genes might be involved in modulating immune cells and/or tumour progression. Taken together, these studies have important implications for designing specific and effective anti-tumour immune therapy strategies that involve manipulation of tumour cells, dendritic cells and regulatory T cells. / Medicine, Faculty of / Medicine, Department of / Experimental Medicine, Division of / Graduate

Vinclozolin (VCZ), an antiandrogenic fungicide, is an endocrine disrupting chemical that is known to possess high affinity for the androgen receptor (AR) and modulate expression of critical androgen-dependant genes in the prostate. In this study, viability and expression of AR, NKX3.1 and CYP3A4 genes were measured in androgen-sensitive prostate cells LNCaP after exposure to VCZ and VCZ treated with S9 microsomes in a time and dose dependent manner. NKX3.1 is an androgen regulated gene that plays a vital role in prostate development. CYP3A4 is involved in xenobiotic metabolism. VCZ decreased the viability at high doses after 48 hours which was slightly mitigated by treatment with S9 metabolites. Expression of NKX3.1 and CYP3A4 was upregulated while an initial downregulation of AR was observed. NKX3.1 upregulation corroborates with possibility of antiandrogens to act as androgens in LNCaP. The results illustrate that VCZ can interfere with the expression of critical prostate genes.

Le phénomène métastatique est important à comprendre puisque de manière irrémédiable, une fois engagé, il conduit le plus souvent au décès des patients. Le cancer de la prostate représente un bon modèle car sa progression du stade hormono-dépendant vers le stade d'hormono-échappement s’accompagne par l’apparition de métastases. Les Protease Activated Receptors (PAR1-4) sont des récepteurs qui jouent un rôle important dans l'hémostase et l'inflammation et dont l’implication dans la prolifération et l'invasion des cellules tumorales a été décrite dans plusieurs tissus. L’étude comparative de l'expression in vitro de PAR1 a confirmé son rôle dans la prolifération et l'invasion des lignées prostatiques normales et tumorales hormono-sensibles comme hormono-indépendantes. Son expression in vivo dans des tissus prostatiques à différents stades pathologiques a montré une surexpression de PAR1 chez les patients ayant atteint le stade d'hormono-échappement, associée à un mauvais pronostic. Son absence s’avère par contre de bon pronostic chez les patients hormonodépendants. L'ensemble des résultats obtenus nous permet de proposer PAR1 comme un nouveau marqueur pronostique pour le cancer de la prostate. L’activation des PARs, comme celle de plusieurs récepteurs de chémokines, apparaît comme un élément fondateur de la transition vers l’état métastatique. Le décryptage de cette combinatoire permettra de mieux comprendre les phénomènes impliqués dans cette transition et permettra de développer des thérapies ciblées pour prévenir l’apparition délétère de métastases / Metastasis is nowadays an important field of research as, once engaged, patients will generally die from their metastatic cancer. Prostate cancer represents an interesting model as its progression from hormone-naïve to hormone-independent status lead to metastatic disease. Protease Activated Receptors (PAR1-4) are G-protein-coupled receptors that play crucial roles in blood coagulation and inflammation but that are likely to play fundamental role in tumor cells proliferation and invasion. In vitro analysis of PAR1 expression in prostate cancer cell lines has confirmed the role of PAR1 in prostate cancer proliferation and invasion. Its expression in vivo in prostate cancer tissues have shown a constant surexpression in hormonerefractory ones, associated with a worse prognosis. However, its absence in hormone-naïve tissues is associated with a good prognosis. These results prompted us to recommend PAR1 as a new prognostic marker associated with prostate cancer progression. PAR activation, as well as several chemokine receptors, seems to be a founder feature of cancer transition to metastasis. Deciphering the pattern of receptor activation will allow a better understanding of the events that drive transition to metastasis and thus the development of new specific targeted therapeutics aimed at stopping deleterious metastatic evolution

Orientadores: Eduardo Tavares Costa, Paulo José Cecílio / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-24T11:03:55Z (GMT). No. of bitstreams: 1 Biazotto_Bruna_M.pdf: 4075478 bytes, checksum: 077630eb4f66ca28ec46de7251c31e3f (MD5) Previous issue date: 2013 / Resumo: A experiência clínica em tratamentos radioterápicos de neoplasia de próstata baseia-se no cálculo de doses em meios homogêneos. Entretanto, o feixe de radiação atravessa tecidos de densidades eletrônicas diferentes como os ossos, que alteram a distribuição de dose. Com o advento da tomografia computadorizada e de algoritmos mais avançados que modelam o feixe de radiação, as heterogeneidades entre os tecidos podem ser incorporadas nos planejamentos de tratamentos radioterápicos. Todavia, não há consenso se as alterações na dose por correções de heterogeneidade são significativas. Por tais razões, pretendeu-se no presente trabalho avaliar a necessidade das correções de heterogeneidade em planejamentos de tratamentos radioterápicos de câncer de próstata. Para isso, analisaram-se as médias das diferenças percentuais nas doses em volume alvo e órgãos de risco obtidas em cálculos com e sem correções de heterogeneidade utilizando imagens tomográficas reais de pacientes que trataram dessa neoplasia. Essa avaliação foi realizada para dois métodos de tratamentos diferentes. O primeiro é o conformacional tridimensional (25 casos), algoritmos de cálculo Convolution, Superposition e Fast Superposition do sistema de planejamento XiO/Elekta, feixes de 6 e 10 MV e 4 campos em box. O segundo por intensidade modulada (14 casos), algoritmo de cálculo Pencil Beam Convolution do sistema de planejamento Eclipse/Varian com dois métodos de correção Batho Modificado e Razão Tecido-Ar Equivalente, feixe de 6 MV e geometria de 5 campos oblíquos. As diferenças percentuais médias resultantes nos volumes estudados foram menores que a incerteza aceita atualmente no cálculo de dose de 3% para as duas modalidades de tratamento. Apesar disso, a variabilidade na anatomia dos pacientes, geometria de campos e energia dos feixes apontam para a necessidade de tais correções e a utilização de métodos ainda mais exatos para a diminuição dessa incerteza no futuro / Abstract: Clinical experience in radiotherapy treatments for prostate cancer is based on the calculation of doses in homogeneous media. However, the radiation beam traverses different electron densities in tissues such as bone, altering the dose distribution. With the advent of computed tomography and more advanced algorithms that model the radiation beam, the heterogeneity between tissues can be incorporated in the planning of radiotherapy treatments. However, there is no consensus whether changes in dose for inhomogeneity corrections are significant. For these reasons, this study intended to evaluate the need for inhomogeneity corrections in treatment planning for radiotherapy of prostate cancer. We have analyzed the average percentage differences in doses in the target volume and organs at risk obtained by calculations with and without heterogeneity corrections using actual CT images of patients treated for this cancer. This evaluation was performed for two different methods of treatments. The first is the three-dimensional conformational (25 cases), calculation algorithms Convolution, Superposition and Fast Superposition from the computerized planning system XiO/Elekta, beams of 6 and 10 MV and 4 fields in box. The second by intensity modulated (14 cases), calculation algorithm Pencil Beam Convolution from the computerized planning system Eclipse/Varian with two correction methods Modified Batho and Equivalent Tissue-Air Ratio, 6 MV beam and geometry of 5 oblique fields. The resulting average percentage differences in volumes studied were smaller than the currently accepted uncertainty in the dose calculation of 3% for both treatment modalities. Nevertheless, variability in anatomy of patients, geometry and field energy beams brings the need for these corrections and the use of more accurate methods to reduce this uncertainty in the future / Mestrado / Engenharia Biomedica / Mestra em Engenharia Elétrica

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The prostate is prone to three pathological processes that include inflammation, benign prostate hyperplasia (BPH) and tumors. According to the center for Disease and Control 1999-2012 report, prostate cancer is the second leading cause of death in the United States. Scientific evidence suggests that up to 30% of men in the general population aged from 50 years and above, irrespective of geographic origin, have foci of prostate neoplastic growth. Unbalanced ROS production and a dysregulated antioxidant defence system have been implicated in prostate cancer development. The transformation of a normal cell into cancer takes a very long period. This observation provides the advantage of using nutraceuticals to prevent, arrest or reverse the cellular and molecular processes of carcinogenesis. Based on scientifically observed positive health roles of green tea (Cameli sinensis) and rooibos (Aspalathus linearis) on major diseases like atherosclerosis, hepatitis and certain types of cancer, this thesis evaluated the effects of these two teas on benign (RPWE 1) and malignant (LNCaP) prostate cells. This was done through the quantification of reactive oxygen species (ROS) using a fluorescence dye 5,6 CM-H2DCFDA, total prostate specific antigen (PSA) levels using a PSA ELISA kit, cell viability using the MTT assay, apoptosis using Tali annexin V stain and cell imaging studies using a Zeiss axiovert 200M inverted fluorescence microscope. Statistical analysis was done using graphpad prism. The findings of this study show that aqueous extracts of green and black tea, fermented and unfermented rooibos and their active compounds epigallocatechin gallate (EGCG) and aspalatin, respectively, are cytotoxic in malignant (LNCaP) prostate cells but exert protective effects in benign (RPWE 1) prostate cells. This thesis implicates the pro-oxidant and anti-oxidant properties of the plant extracts, respectively, for the above mentioned effects. In this regard, tea and rooibos promoted ROS production in malignant (LNCaP) prostate cells, which subsequently promoted cell death of the malignant cells through apoptosis and necrosis. Further to this, tea and rooibos used in this thesis, protected normal prostate cells from the adverse effects of ROS. In this regard, fluorescence microscope photographs showed RPWE 1 cells with low DCF fluorescence compared to the malignant prostate cells. Low magnification light microscope photographs showed RPWE 1 cells with flat polygonal shapes and increased adherence both at low and high concentrations of tea and rooibos. On the contrary, high concentrations of tea and rooibos on malignant (LNCaP) prostate cells induced stress, which made the cells attain irregular shapes and as the stress levels increased, cells became detached and appeared dead. Flow cytometry confirmed the presence of apoptotic and necrotic cell in malignant (LNCaP) prostate cells. In this thesis, EGCG and aspalathin were responsible for the high rates of apoptosis observed whereas green tea and unfermented rooibos induced the highest rate of necrosis. Further to this, tea and rooibos and the main active compounds EGCG and aspalathin, respectively, significantly promoted the reduction of total serum prostate specific antigen (PSA) in malignant prostate cells. In normal prostate cells, these plant extracts maintained the total serum PSA at its basal physiological level. In this thesis, to the best of our knowledge, we report for the first time the cell-specific effects of fermented rooibos, unfermented rooibos and their main active component aspalathin, on prostate cancer cells. We showed that rooibos and aspalathin exert pro-oxidant effects on malignant LNCaP cells and anti-oxidant effects on benign RPWE 1 cells. In conclusion, tea (C. sinensis) and rooibos (A. linearis) and their respective main active compounds, epigallocatechin gallate and aspalathin, are cytotoxic to malignant prostate cells whereas in normal prostate cells, they have protective effects against ROS induced stress. The pro-oxidant and anti-oxidant effects are responsible for the aforementioned effects respectively. The decrease in total serum PSA demonstrate the strong therapeutic effects that tea and rooibos have on malignant (LNCaP) prostate cells. / Malawi Government: Department of Human Resources Development and Management

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Prostate cancer is one of the heterogeneous groups of neoplastic diseases originating from the reproductive system of the male naming, the prostate gland. In the west, prostate cancer is the most common cancer affecting African men in older age (over the age of 55) and usually with a family history of the disease. The initiation and progression of this disease is thought to result from the genetic alterations of gene expression in the prostate epithelial cells. Prostate cancer has a very slow progression. This observation provides the advantage of early detection and the notion for using diet to prevent the cellular and molecular processes of carcinogenesis. Epidemiological research has documented a positive health role for red palm oil on atherosclerosis, arterial thrombosis and several types of cancers. This thesis focuses on investigating the effect of different concentrations of the red palm oil (0.1, 1, 10, 100, 500, 1 000 μg/ml) on malignant (LNCaP) prostate cells and benign (PWR-1E) prostate cells over 24 and 72- hours. The following parameters were investigated: cell morphology and viability (using MTT assay), the expression of androgen receptors and prostate-specific antigen (PSA) via RT-PCR and/or PSA ELISA kit. The results of this study demonstrate that red palm oil has significant cytotoxic effects on malignant (LNCaP) prostate cells but caused only a slight decrease in cell viability of benign (PWR- 1E) prostate cells. Morphologically, we noted a clear increase in detachment and cell death in malignant (LNCaP) cells as the concentrations of red palm oil increased. Moreover, the viability decreased significantly in both 24 and 72-hour treatment of red palm oil. Further to this, red palm oil significantly promoted the reduction of total PSA concentration in malignant (LNCaP) prostate cells whereas in benign (PWR-1E) prostate cells the Red Palm Oil maintained the total serum PSA at its basal physiological level. In conclusion, red palm oil is significantly cytotoxic to malignant (LNCaP) prostate cells whereas weakly cytotoxic effect toward benign (PWR-1E) prostate cells. The potent inhibition to mitochondrial dehydrogenase activity is responsible for the aforementioned effects respectively. The decrease in total serum PSA demonstrate the strong therapeutic effects that red palm oil has on malignant (LNCaP) prostate cells

Background and Aim: Prostate cancer (PCa) is the leading cancer diagnosis amongst South African men. The incidence of PCa is 68.0 per 100 000 Age Standardized Rate (ASR) and the mortality rates are 27.9 per 100 000 ASR; Globocan 2018. Diagnosis of PCa is based on a combination of digital rectal examination, prostate-specific antigen (PSA) and histology. Several biomarkers have been used to increase the sensitivity and specificity of PSA in distinguishing patients with PCa from those with benign prostatic hyperplasia (BPH). These include fractionated PSA, free/total PSA ratio, −2proPSA, prostate cancer antigen 3 and prostate health index amongst others. Biomarkers are needed to differentiate BPH from PCa due to a lack of specificity of these markers with PSA levels above 4.0 ng/ml. The aim of this study is to investigate gene expression patterns of South African men in 9 PCa and 10 BPH patients in order to distinguish between the two groups. Methods: Ethical approval was obtained (HREC 454/2012). Patients scheduled for transurethral resection of the prostate were recruited from the Western Cape. RNA was extracted from prostate tissue using the AllPrep DNA/RNA/miRNA Universal Kit (Qiagen). Complementary DNA was synthesized from RNA using the SuperScript IV VILO Master Mix (Thermo Fischer Scientific). Gene expression was analyzed with the Human Prostate Cancer RT2 Profiler PCR Array and SYBR Green Master Mix. Data were analyzed with the GeneGlobe RT2 and miScript PCR Array Data Analysis Centre from Qiagen. Results: The cohort included patients from different ethnic groups namely, Caucasians, Mixedand African ancestry. The PCa group has an age range from 56 to 75 years (mean 65) while the BPH group was slight older ranging from 60 to 76 years (mean 68). PSA levels range from 24 to 5000 ng/ml (mean 1252 ng/ml, median 185) for the PCa group and 11 to 58 ng/mL (mean 25 ng/ml, median 22) for the BPH group. The following genes were downregulated 2-fold in the PCa group with p values s <0.05; IGF1, PTEN, GSTP1, SOCS3, EGR3, GPX3, TIMP3, ZNF185, DKK3, PTGS2, FOXO1, ARNTL, TNFRSF10D, CCND1, and DLC1, upregulated genes included; CDH1, MKI67, TMPRSS2, ERG, CDKN2A, FASN, and AR but were not statistically significant. At a fold change threshold of 1.5, the following additional genes were downregulated in the PCa group with p values <0.05; DAXX, EGFR, RASSF1, SOX4, and TIMP2, upregulated genes were ACACA, AR, CDKN2A, ERG and FASN but were also not statistically significant. The study shows similarly differentially expressed genes as seen in international studies. Of note PTEN, MKI67 and FASN which are associated with poor prognosis. EGR3 was downregulated in our study and this has been associated aggressive disease and predict relapse after PCa treatment. This could explain the high mortality demonstrated in South African epidemiological studies. Conclusion: We identified a group of differentially expressed genes that have potential in distinguishing PCa and BPH patients with PSA values above 10 ng/ml. A larger population study is needed to further evaluate the clinical significance of our findings.

LIMK2 is a serine/threonine/tyrosine kinase that promotes tumor cell invasion and metastasis by phosphorylating cell proteins and altering their functions. There is a need to find tumor-specific substrates for LIMK2 in order to understand the downstream pathway of these substrates, their function, and how they are regulated by LIMK2. Recently, our labrotory identified LIMK2 as an excellent target for curing castration-resistant prostate cancer (CRPC). In this study, we identify two novel substrates for LIMK2 in CRPC: speckle-type POZ protein (SPOP), and Y-box binding protein-1 (YBX1). While LIMK2 negatively regulates SPOP, it positively regulates YBX1 − both by phosphorylation using in-vitro kinase assays. A study in our labrotory also proved that LIMK2 regulates Aurora A kinase (AURKA), where AURKA directly phosphorylates LIMK2. AURKA is a serine/threonine kinase that regulates cell cycle during mitosis; it is known to be upregulated, with uncontrolled activity, in many types of cancer, including prostate cancer. It is therefore important to identify new substrates for AURKA, especially in light of reported lethality in early embryonic mice, in association with AURKA-knockout. In other words, targeting AURKA directly may cause severe toxicity, a finding that has prevented direct inhibitors from passing Phase II clinical trials. In this study, we also identified SPOP and YBX1 as direct substrates for AURKA. Our results confirm what we know about the LIMK2/AURKA relationship: that AURKA negatively regulates SPOP and positively regulates YBX1. Targeting LIMK2 and AURKA indirectly through SPOP, YBX1 and its other substrates holds tremendous therapeutic potential in treating prostate cancer. With this, we open the door for researches to investigate the direct phosphorylation of SPOP and YBX1 in other types of cancer cells known to have overexpression in SPOP and/or YBX1.

One of the most common types of cancer among women is breast cancer. It represents one of the diseases leading to a high number of mortalities among women. On the other hand, prostate cancer is the second most frequent malignancy in men worldwide. The early detection of prostate cancer is fundamental to reduce mortality and increase the survival rate. A comparison between six types of machine learning models as Logistic Regression, Decision Tree, Random Forest, Gradient Boosting, k Nearest Neighbors, and Naïve Bayes has been performed. This research aims to identify the most efficient machine learning algorithms for identifying the most significant risk factors of prostate and breast cancers. For this reason, National Health Interview Survey (NHIS) and Prostate, Lung, Colorectal, and Ovarian (PLCO) datasets are used. A comprehensive comparison of risk factors leading to these two crucial cancers can significantly impact early detection and progressive improvement in survival. / Includes bibliography. / Thesis (P.S.M.)--Florida Atlantic University, 2021. / FAU Electronic Theses and Dissertations Collection