雷公藤甲素通过抑制microRNA-21的表达提高SGC7901/CDDP细胞顺铂敏感性

Triptolide enhances the sensitivity of SGC7901/CDDP cells to cisplatin via inhibiting microRNA-21 expression

目的:观察雷公藤甲素对胃癌SGC7901/CDDP细胞顺铂敏感性的影响,并探讨其机制。方法:采用非毒性浓度雷公藤甲素作用于SGC7901/CDDP细胞,四甲基偶氮唑盐比色法(MTT法)检测细胞增长率;荧光定量PCR检测microRNA-21表达;蛋白质印迹法检测Bcl-2蛋白表达;AnnexinV/PI双标法检测细胞凋亡。将microRNA-21反义寡核苷酸转染SGC7901/CDDP细胞,观察其对细胞增长率、凋亡和Bcl-2表达的影响。采用小干扰RNA转染实验证明Bcl-2与胃癌耐药的关系。结果:非毒性浓度雷公藤甲素可提高SGC7901/CDDP细胞对顺铂敏感性,并且降低mi-croRNA-21的表达。microRNA-21反义寡核苷酸转染SGC7901/CDDP细胞后,降低Bcl-2表达,提高顺铂敏感性和顺铂诱导的细胞凋亡。小干扰RNA实验证明Bcl-2水平与SGC7901/CDDP细胞顺铂耐药有关。结论:雷公藤甲素通过降低microRNA-21的表达,抑制Bcl-2蛋白表达,促进顺铂诱导的细胞凋亡,从而提高胃癌SGC7901/CDDP细胞顺铂敏感性。

Objective： To investigate the effect of triptolide on the sensitivity of SGC7901/CDDP cells to cisplatin and its mechanism.Methods： SGC7901/CDDP cells were treated with triptolide at non-toxicity concentration.The cell growth was detected using MTT method.The expression of microRNA-21 was measured by Fluorescence quantitative polymerase chain reaction.Bcl-2 protein level was determined by Western blotting.The apoptosis of cells was measured by Annexin V/PI methods.After transfection with microRNA-21 antisense oligonucleotide,the sensitivity to cisplatin,the apoptosis of cells and Bcl-2 protein level were detected in SGC7901/CDDP cells.Small interference RNA assay was used to determine the relationship of Bcl-2 and drug resistance in SGC7901/CDDP cells.Results： Triptolide at non-toxicity concentration significantly enhanced the sensitivity of SGC7901/CDDP cells to cisplatin and inhibited the expression of microRNA-21.After transfected with microRNA-21 antisense oligonucleotide,Bcl-2 protein level was decreased in SGC7901/CDDP cells.MicroRNA-21 antisense oligonucleotide enhanced the sensitivity to cisplatin and the apoptosis induced by cisplatin.Bcl-2 played an important role in the resistance of SGC7901/CDDP cells to cisplatin.Conclusions： Triptolide enhanced the sensitivity of SGC7901/CDDP cells to cisplatin via inhibiting microRNA-21 expression and decreasing the Bcl-2 level.