Adenovirus-mediated expression of p33 ING1b induces apoptosis and inhibits proliferation in gastric adenocarcinoma cells in vitro

摘要

Background Inhibitor of growth 1b (ING1b) is considered to be a class II tumor suppressor gene. Although reduced expression of p33 ING1b has been reported in many human malignancies, including gastric cancers, the effect of p33 ING1b on gastric cancer cells has yet to be investigated. Methods Expression of p33 ING1b in gastric adenocarcinoma tissues and their adjacent non-malignant gastric mucosa, as well as in gastric adenocarcinoma cell lines and normal gastric epithelial cells, was detected by using Western blotting. Recombinant adenoviruses were prepared to mediate the ectopic expression of p33 ING1b (Ad-ING1b) and green fluorescent protein (GFP)(Ad-GFP) in the gastric adenocarcinoma cell lines, SGC-7901, MKN28, and MKN45 and the normal gastric epithelial cell line GES-1. Alterations in the proliferation and apoptosis of the cells after adenoviral infection were determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively, and cell cycle distribution was analyzed in a fluorescenceactivated cell sorter. Results Western blotting confirmed the reduced expression of p33 ING1b in gastric adenocarcinoma tissues and gastric adenocarcinoma cell lines. The ectopic expression of p33 ING1b mediated by Ad-ING1b resulted in decreased growth, increased apoptosis, and cell cycle arrest at the G1 phase in both benign and malignant gastric epithelial cells regardless of their p53 status. Addition of a p53 inhibitor, pifithrin-a, did not abolish the pro-apoptotic and cell cyclearresting effects of p33 ING1b in p53 wild-type cells. Conclusions Down-regulation of p33 ING1b might play an important role in the development of gastric adenocarcinoma. Targeted local expression of p33 ING1b may offer a promising alternative therapeutic measure for gastric cancer.