Predictive markers for the response to 5-fluorouracil therapy in cancer cells: Constant-field gel electrophoresis as a tool for prediction of response to 5-fluorouracil-based chemotherapy

Abstract

Abstract. The prediction of response or severe toxicity and therapy individualisation are extremely important in cancer chemotherapy. There are few tools to predict chemoresponse or toxicity in cancer patients. We investigated the correlation between the induction and repair of DNA double-strand breaks (DSBs) using constant-field gel electrophoresis (CFGE) and evaluating cell cycle progression and the sensitivity of four cancer cell lines to 5-fluorouracil (5FU). Using a sulphorhodamine-B assay, colon carcinoma cells (HCT116) were found to be the most sensitive to 5FU, followed by liver carcinoma cells (HepG2) and breast carcinoma cells (MCF-7). Cervical carcinoma cells (HeLa) were the most resistant. As measured by CFGE, DSB induction, but not residual DSBs, exhibited a significant correlation with the sensitivity of the cell lines to 5FU. Flow cytometric cell cycle analysis revealed that 14% of HCT116 or HepG2 cells and 2% of MCF-7 cells shifted to sub-G1 phase after a 96-h incubation with 5FU. Another 5FU-induced cell cycle change in HCT116, HepG2 and MCF-7 cells was the mild arrest of cells in G1 and/or G2/M phases of the cell cycle. In addition, 5FU treatment resulted in the accumulation of HeLa cells in the S and G2/M phases. Determination of Fas ligand (Fas-L) and caspase 9 as representative markers for the extrinsic and intrinsic pathways of apoptosis, respectively, revealed that 5FU-induced apoptosis in HCT116 and HepG2 results from the expression of Fas-L (extrinsic pathway). Therefore, the induction of DNA DSBs by 5FU, detected using CFGE, and the induction of apoptosis are candidate predictive markers that may distinguish cancer cells which are likely to benefit from 5FU treatment and the measurement of DSBs using CFGE may aid the prediction of clinical outcome.

Efficacy and toxicity of different concurrent chemoradiotherapy regimens in the treatment of advanced cervical cancer

“…Additionally, FU is commonly used to treat skin cancer or damage caused by the HPV virus so that it benefitted CC caused by the HPV virus. [41] Previous studies had also confirmed that if FU was used alone, the curative rate was approximately 75%; therefore, it was better to combine it with other drugs. [42] This supported our finding that the 5-year OS rate of CCRT (cisplatin + FU + hydroxyurea) was relatively higher than the other CCRT regimens in our study.…”

Mechanism of 5-Fluorouracil-Induced Apoptosis on Cultured Corneal Endothelial Cells

“…The intrinsic pathway relies on extrinsic signals to promote cell death in a cell-and tissue-dependent manner following DNA damage [20]. Evidence has shown that treatment with 5-FU increased the expression of Fas, which belongs to the death receptor pathway [21]. In normal thymus, 5-FU induced apoptosis is related to Fas, Bax, and caspase 3 coexpression [22].…”

Proliferation, Apoptosis, and Invasion Effects of Mistletoe Alkali on Human Osteosarcoma U2OS In Vitro

“…5-FU is reported to induce the apoptosis of colon carcinoma cells (HCT116), liver carcinoma cells, and breast carcinoma cells (MCF-7) and is therefore used in anticancer chemotherapy. 33 It has been previously shown that a number of different mistletoe extracts can trigger apoptosis in various cancer cell lines. 24,34,35 In this study we used both TUNEL and ICC staining of caspase 3 to detect apoptosis.…”