Medical Oncology

, Volume 27, Issue 2, pp 397–405

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

  • Anning Yin
  • Yingan Jiang
  • Xianfeng Zhang
  • Hesheng Luo
Original Paper

DOI: 10.1007/s12032-009-9224-x

Cite this article as:
Yin, A., Jiang, Y., Zhang, X. et al. Med Oncol (2010) 27: 397. doi:10.1007/s12032-009-9224-x

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.

Keywords

Fas-associated death domain protein (FADD)5-FluorouracilApoptosisColorectal adenocarcinoma

Copyright information

© Humana Press Inc. 2009

Authors and Affiliations

  • Anning Yin
    • 1
  • Yingan Jiang
    • 1
  • Xianfeng Zhang
    • 1
  • Hesheng Luo
    • 1
  1. 1.Department of GastroenterologyRenmin Hospital of Wuhan UniversityWuhanPeople’s Republic of China