BAK overexpression mediates p53-independent apoptosis inducing effects on human gastric cancer cells

BMC Cancer
Qiang-Song TongWei Qian

Abstract

BAK (Bcl-2 homologous antagonist/killer) is a novel pro-apoptotic gene of the Bcl-2 family. It has been reported that gastric tumors have reduced BAK levels when compared with the normal mucosa. Moreover, mutations of the BAK gene have been identified in human gastrointestinal cancers, suggesting that a perturbation of BAK-mediated apoptosis may contribute to the pathogenesis of gastric cancer. In this study, we explored the therapeutic effects of gene transfer mediated elevations in BAK expression on human gastric cancer cells in vitro. Eukaryotic expression vector for the BAK gene was constructed and transferred into gastric cancer cell lines, MKN-45 (wild-type p53) and MKN-28 (mutant-type p53). RT-PCR and Western Blotting detected cellular BAK gene expression. Cell growth activities were detected by MTT colorimetry and flow cytometry, while apoptosis was assayed by electronic microscopy and TUNEL. Western Blotting and colorimetry investigated cellular caspase-3 activities. BAK gene transfer could result in significant BAK overexpression, decreased in vitro growth, cell cycle G0/G1 arrest, and induced apoptosis in gastric cancer cells. In transferred cells, inactive caspase-3 precursor was cleaved into the active subunits p20...Continue Reading

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Citations

Mar 3, 2007·Apoptosis : an International Journal on Programmed Cell Death·Sonish AzamDindial Ramotar
Mar 29, 2014·BioMed Research International·Naglaa Mohamed KholoussiAhmed F El-Kased
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Methods Mentioned

BETA
PCR
electrophoresis
flow cytometry
transfection

Software Mentioned

CELLQEST
SPSS

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