外源性FHIT基因表达对顺铂诱导宫颈癌SiHa细胞凋亡的影响

Combined effects of extopic FHIT gene with cisplatin on SiHa apoptosis.

目的:探讨外源性脆性组氨酸三联体(fragile histidine triad,FHIT)基因表达与顺铂(cisplatin,DDP)是否可协同促进宫颈癌SiHa细胞的凋亡。方法:重组质粒PRcC-MV-FHIT(A组)及空质粒PRcCMV(B组)分别转染无内源性FHIT蛋白表达的SiHa细胞(未转染SiHa细胞为C组),Western Blot检测外源性FHIT蛋白的表达,使用不同浓度的顺铂分别处理各组细胞,通过AO-EB染色、流式细胞术检测顺铂处理后各组细胞的凋亡。结果:顺铂处理48h后进行流式细胞仪检测,重组质粒+DDP2.0μg/ml、未转染细胞+DDP2.0μg/ml及未用顺铂处理的重组质粒组凋亡率分别为51.03%、20.58%、22.07%,重组质粒+DDP组细胞凋亡更明显(重组质粒+DDP2.0μg/ml组与未转染细胞+DDP2.0μg/ml及未用顺铂处理的重组质粒组相比P<0.01),AO-EB染色亦见顺铂处理的重组质粒组凋亡细胞明显增多。结论:外源性FHIT基因表达与顺铂协同促进SiHa细胞凋亡,为基因治疗与化疗联合治疗宫颈癌提供理论基础。

Objective： To investigate the effect of fragile histidine triad （FHIT） expression on the apoptosis induced by chemotherapeutics agent cisplatin （DDP）. Methods： PRcCMV-FHIT and PRcCMV were transfected into cervical cancer cell line SiHa lacking fulllength FHIT transcripts and endogenous FHIT protein, the expression of extopic FHIT protein was examinated by Western blot,apoptosis were examinated by AO-EB stain and flour cell assess（FCA）. Results： The apoptosis rate was respectively 51.03% ,20.58% ,22.07% in recombine plasmid ＋ DDP2.01μg/ml group,blank cell ＋ DDP2.0μg/ml group,recombine plasmid group,the number of apoptotic cells was also statistically increased in recombine plasmid ＋ DDP2.01μg/ml group by AO-EB stain. Conclusion： Cisplatin-induced apoptosis can be enhanced by extopic FHIT expression in cervical cancer cell line SiHa, which provides a experimental basis for the treatment of cervix neoplasms with gene therapy combined with chemotherapy.