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Searching for Radiosensitizers: Development of a Novel Assay and High-throughput ScreeningKatz, David 24 February 2009 (has links)
The colony formation assay (CFA) is the gold standard for measuring cytotoxic effects on cells. To increase efficiency, the CFA was converted to a 96-well format using an automated colony counting algorithm. The 96-well CFA was validated using ionizing radiation (IR) on the FaDu and A549 cancer cell lines. Its ability to evaluate combination therapies was investigated using cisplatin and IR. The 96-well CFA was transferred to a robotic platform for evaluation as a high-throughput screen (HTS) readout for the discovery of novel anti-cancer compounds, and radiosensitizers. Screening yielded eight putative anti-cancer hits, and five putative radiosensitizing hits. Secondary screening confirmed 6/8 anti-cancer compounds, and 0/5 radiosensitizing compounds. Thus, the 96-well CFA can be adopted as an alternative assay to the 6-well CFA in the evaluation of cytotoxicity in vitro, providing a possible readout to be utilized in HTS for discovering anti-cancer compounds, but with limited applicability in discovering radiosensitizers.
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Searching for Radiosensitizers: Development of a Novel Assay and High-throughput ScreeningKatz, David 24 February 2009 (has links)
The colony formation assay (CFA) is the gold standard for measuring cytotoxic effects on cells. To increase efficiency, the CFA was converted to a 96-well format using an automated colony counting algorithm. The 96-well CFA was validated using ionizing radiation (IR) on the FaDu and A549 cancer cell lines. Its ability to evaluate combination therapies was investigated using cisplatin and IR. The 96-well CFA was transferred to a robotic platform for evaluation as a high-throughput screen (HTS) readout for the discovery of novel anti-cancer compounds, and radiosensitizers. Screening yielded eight putative anti-cancer hits, and five putative radiosensitizing hits. Secondary screening confirmed 6/8 anti-cancer compounds, and 0/5 radiosensitizing compounds. Thus, the 96-well CFA can be adopted as an alternative assay to the 6-well CFA in the evaluation of cytotoxicity in vitro, providing a possible readout to be utilized in HTS for discovering anti-cancer compounds, but with limited applicability in discovering radiosensitizers.
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Mathematical models of budding yeast colony formation and damage segregation in stem cellsWang, Yanli January 2017 (has links)
No description available.
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The effect of RAN inhibition on human colorectal cancer cells (CRC)Elrewey, Hussein A.S. January 2020 (has links)
Colorectal cancer (CRC) is the third most widespread and fourth most fatal
malignancy disease. The CRC from a primary site can spread to other tissues,
forming secondary tumours. CRC can metastasise to the liver through the effect
of K-Ras and Pten mutation (Mt.) (Abbas et al. 2020). This study aimed to assess
the hypothesis that the Ran inhibitor mebendazole MBZ reduces cell invasion
and metastasis of CRC. I have investigated MBZ effect on the CRC isogenic
human cell lines with specific mutations (HCT-116 K-Ras, DLD-1 K-Ras and Pten
deletion and wild type HCT-116 and DKO-3. I used qRT-PCR and western
blotting to identify expression levels of various genes and signalling molecules
after treatment with 0.5 mM MBZ. In addition, several assays were performed to
investigate MBZ effect on biological properties of the cells such as proliferation,
migration, invasion, and colony formation. MBZ downregulated Ran and induced
apoptosis through inhibition of Bcl-2 expression as well as inducing caspase -3,
-7, -9 and PARP cleavage. Moreover, MBZ showed an effect on immune
response by down regulating C5a, IL-1ß and IL-1α analysed at mRNA level.
When treated with MBZ, the migration, invasion and colony formation abilities of
HCT-116 K-Ras Mt., DLD-1 K-Ras Mt. and HCT-116 Pten-/- were significantly
reduced compared to a control treated cell line. This was also the case with wild type cell lines such as HCT-116 and DKO-3. Furthermore, signalling molecules
such as p- Erk 1/2 and p- Akt were upregulated after MBZ treatment and exert
inhibition on Akt 1/2/3 and VEGFR1/2 mRNA levels. In conclusion; MBZ which is
a Ran inhibitor, has significantly reduced proliferation, colony formation, and
migration in colorectal cell lines with K-Ras and Pten gene deletion compared to
wild type cells in a dose-dependent manner. This work paves the way to clinical
validation of MBZ as a combination therapy for reducing the invasion of CRC
cells.
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Cytokine Profiles of Head and Neck Squamous Cell Carcinoma Undergoing Dual Immunotherapy With Cetuximab and Pembrolizumab Identify Interferon Gamma-Induced Protein 10 as Novel BiomarkerBerszin, Michael, Michaelides, Ioannis, Siemert, Julia, Röhl, Louisa, Wellhausen, Jana, Wald, Theresa, Bohr, Christopher, Künzel, Julian, Gradistanac, Tanja, Dietz, Andreas, Zebralla, Veit, Pirlich, Markus, Wiegand, Susanne, Wichmann, Gunnar 05 April 2023 (has links)
Background: Pembrolizumab and cetuximab are antibodies under investigation in head
and neck squamous cell carcinoma (HNSCC) either as single agents or combined with
cisplatin and other chemotherapeutic drugs, e.g., 5-fluorouracil and/or docetaxel.
However, also the combination of both antibodies may have potential in recurrent/
metastatic (R/M) HNSCC, in particular in cisplatin-resistant or -refractory cases or
patients with comorbid disease, e.g. patients with impaired renal function.
Methods: To clarify potential benefit that may result from such combination, we used the
FLAVINO assay, a short-time ex vivo assay to compare responsiveness of HNSCC to
pembrolizumab, cetuximab and both combined regarding colony formation of epithelial
cells of biopsy-derived tumor samples and their cytokine production within three days
either without or with stimulation with 10 ng/mL interferon gamma (IFN-g). Vascular
endothelial growth factor A (VEGF), monocyte chemoattractant protein 1 (MCP-1 or
CCL2), interleukin 6 (IL-6), IL-8, IFN-g, and interferon gamma-induced protein 10 (IP-10 or
CXCL10) in supernatants were measured by ELISA.
Results: We detected huge heterogeneity in response to cetuximab, pembrolizumab and
both combined with and without IFN-g stimulation. Moreover, we detected a link between
IFN-g induced IP-10 release and improved outcome in those HNSCC patients who were
capable to respond to IFN-g and pembrolizumab, cetuximab and both combined with a
further increase in IP-10 production. We derived an “IP-10 score” that independent from
clinical characteristics of HNSCC patients and therapy regimens applied was able to
predict their outcome.
Conclusions: The heterogeneity in the ex vivo response of cetuximab, pembrolizumab
and both combined with and without IFN-g stimulation identifies subgroups of HNSCC
patients with deviating OS.
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MYCL promotes iPSC-like colony formation via MYC Box 0 and 2 domains / MYCLはMYC Box 0及び2ドメインを介してiPS細胞様コロニーの形成を促進するAkifuji, Chiaki 23 March 2022 (has links)
京都大学 / 新制・課程博士 / 博士(医科学) / 甲第23817号 / 医科博第138号 / 新制||医科||9(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 川口 義弥, 教授 藤田 恭之, 教授 江藤 浩之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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