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

• Main Authors: Liu Jun, Wang Liang, Zheng Li-Duan, Tong Qiang-Song, Qian Wei
• Format: Article
• Language: English
• Published: BMC 2004-07-01
• Series: BMC Cancer
• Subjects: BAK gene; Gastric cancer; Caspase-3; Apoptosis
• Online Access: http://www.biomedcentral.com/1471-2407/4/33

<p>Abstract</p> <p>Background</p> <p>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 <it>in vitro</it>.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>BAK gene transfer could result in significant BAK overexpression, decreased <it>in vitro </it>growth, cell cycle G₀/G₁arrest, and induced apoptosis in gastric cancer cells. In transferred cells, inactive caspase-3 precursor was cleaved into the active subunits p20 and p17, during BAK overexpression-induced apoptosis. In addition, this process occurred equally well in p53 wild-type (MKN-45), or in p53 mutant-type (MKN-28) gastric cancer cells.</p> <p>Conclusions</p> <p>The data presented suggests that overexpression of the BAK gene can lead to apoptosis of gastric cancer cells <it>in vitro</it>, which does not appear to be dependent on p53 status. The action mechanism of BAK mediated apoptosis correlates with activation of caspase-3. This could be served as a potential strategy for further development of gastric cancer therapies.</p>