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