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Overexpression of FADD enhances 5-fluorouracil-induced apoptosis in colorectal adenocarcinoma cells

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Abstract

To investigate the mechanism of enhancing apoptosis-inducing effects of 5-fluorouracil on human colorectal adenocarcinoma cells by stable transfection of extrinsic Fas-associated death domain protein (FADD) gene, both in vitro and in vivo. FADD gene of stable overexpression was determined by reverse transcription polymerase chain reaction (RT-PCR) assay and Western blotting assay. After treatment with 5-fluorouracil as an apoptotic inducer, in vitro cell growth activities were investigated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis and its rates were evaluated by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) assay and flow cytometry of annexin V-FITC/PI staining. To examine the combination therapeutic effect of FADD and 5-fluorouracil, tumor xenograft model was prepared for in vivo study. Compared with SW480 and SW480/neo cells, FADD mRNA and protein levels of SW480/FADD cells were higher. Chemosensitivity and apoptosis rates of SW480/FADD cells were remarkably higher than SW480 and SW480/neo cells when treated with 5-fluorouracil. In in vivo study, overexpression of FADD increased the efficacy of 5-fluorouracil-induced inhibition of tumor growth in nude mice. Stable overexpression of extrinsic FADD gene can conspicuously ameliorate apoptosis-inducing effects of 5-fluorouracil on colorectal adenocarcinoma cells, which is a novel strategy to improve chemotherapeutic effects on colorectal cancer.

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Acknowledgment

This study was supported by a Grant from National Natural Science Foundation of China (NO.30471690).

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Authors and Affiliations

  1. Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei, People’s Republic of China

    Anning Yin, Yingan Jiang, Xianfeng Zhang & Hesheng Luo

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  1. Anning Yin
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Correspondence to Yingan Jiang.

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Yin, A., Jiang, Y., Zhang, X. et al. Overexpression of FADD enhances 5-fluorouracil-induced apoptosis in colorectal adenocarcinoma cells. Med Oncol 27, 397–405 (2010). https://doi.org/10.1007/s12032-009-9224-x

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  • DOI: https://doi.org/10.1007/s12032-009-9224-x

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