外源性FHIT基因表达对长春新碱诱导人胃癌MKN-28细胞凋亡的影响

Effect of exogenous FHIT gene expression on vincristine-induced apoptosis of human gastric cancer cells MKN-28

目的:探讨外源性FHIT基因表达对长春新碱诱导的胃癌细胞凋亡的影响及其分子机制.方法:通过脂质体将重组FHIT基因PRC/CMV质粒和空载体转染到人胃癌细胞MKN-28,Western blot法检测外源性FHIT蛋白的表达;使用不同浓度的长春新碱分别处理各组细胞,MTT法分析细胞增殖,流式细胞术检测细胞的凋亡;Western blot法检测细胞Bcl-2和Bax的表达.结果:转染FHIT基因后,MKN-28细胞检测到FHIT蛋白的表达;长春新碱处理48h后,转染FHIT基因组细胞、转染空载体组细胞及未转染组细胞的凋亡率分别是30.967%±2.122%、11.033%±1.724%、10.733%±1.021%,转染FHIT基因组细胞凋亡更明显(F=142.045,P<0.05);转染FHIT基因组细胞经长春新碱处理后Bcl-2蛋白表达减少,Bax蛋白表达增加.结论:外源性FHIT基因表达可以增强长春新碱诱导的胃癌细胞凋亡,其机制可能与凋亡相关蛋白Bcl-2和Bax表达有关.

AIM： To investigate effect of exogenous fragile histidine triad （FHIT） gene expression on gastric cancer cells MKN-28 apoptosis induced by vincristine as well as its molecular mechanism. METHODS： The recombinant FHIT gene was transfected into human gastric cancer cells MKN-28 through liposomes. The expression of exogenous FHIT gene was detected using western blot. The MKN-28-pRcCMV-FHIT, MKN-28- pRcCMV and MKN-28 cells were treated with vincristine of different concentrations. After treatment with vincristine, the inhibition rates of the cells in 3 groups were examined using MTT assay. The apoptosis of cells was determined by flow cytometry. The expressions of Bcl-2 and Bax were measured using Western blot. RESULTS： The expression of FHIT protein was detected in MKN-28 cells after transfection with FHIT gene. After being for 48 h, the apoptosis treated with vincristine rates of the cells transfected with FHIT gene, the cells transfected with empty vector and the non-transfected cells were 30.967% ± 2.122%, 11.033% ± 1.724% and 10.733% ± 1.021%, respectively. The apoptosis of the cells transfected with FHIT gene was more obvious （F = 142.045, P 〈 0.05）. The expression of Bcl-2 was down-regulated and that of Bax was up-regulated after treatment with vincristine. CONCLUSION： The expression of exogenous FHIT gene can enhance the vincristine-induced apoptosis in gastric cancer cells, which may be related to the expressions of apoptosis-related protein Bcl-2 and Bax.