干扰KIF14基因对子宫内膜癌细胞顺铂耐药的影响

Influence of interfering KIF14 gene on cisplatin resistance of endometrial cancer cells

目的:探讨KIF14基因对子宫内膜癌顺铂耐药细胞增殖、凋亡及顺铂敏感性的影响。方法:RT-qPCR和western-blot检测子宫内膜癌细胞Ishikawa及子宫内膜癌顺铂耐药细胞株Ishikawa/DDP中KIF14水平;将siRNA-KIF14及siRNA-NC转染Ishikawa/DDP细胞作为si-KIF14组和si-NC组,耐药组细胞不做处理,通过克隆形成实验检测干扰KIF14后子宫内膜癌顺铂耐药细胞增殖情况;流式细胞仪检测干扰KIF14后子宫内膜癌顺铂耐药细胞凋亡情况,western-blot检测增殖、凋亡及耐药基因相关蛋白ki-67、Bax、cleaved-Caspase-3、MDR1、P-gp表达水平;取各组细胞加入系列浓度顺铂,MTT法检测干扰KIF14后子宫内膜癌顺铂耐药细胞对顺铂敏感性。结果:与子宫内膜癌细胞Ishikawa相比,KIF14在子宫内膜癌顺铂耐药细胞株Ishikawa/DDP中高表达(P<0.001);与耐药组和si-NC组相比,干扰KIF14后si-KIF14组Ishikawa/DDP细胞克隆形成数目和ki-67蛋白水平下降,细胞凋亡率以及Bax、cleaved-Caspase-3蛋白水平升高,IC50值下降,多药耐药基因MDR1、P-gp蛋白水平下降(均P<0.001)。结论:干扰KIF14表达能够抑制子宫内膜癌顺铂耐药细胞Ishikawa/DDP增殖,促进凋亡,增加对顺铂的敏感性。

Objective:To investigate the influence of KIF14 gene on proliferation,apoptosis,and cisplatin sensitivity of endometrial carcinoma cells with cisplatin resistant.Methods:The expression levels of KIF14 in endometrial carcinoma cell line Ishikawa and cisplatin resistant cell line Ishikawa/DDP were detected by RT-qPCR and Western-blot.The Ishikawa/DDP cells in siRNA-KIF14 group were transfected by siRNA-KIF14 and the Ishikawa/DDP cells in siRNA-NC group were transfected by siRNA-NC,and the cells in drug-resistant group were not treated.The proliferation of cisplatin resistant cells of endometrial carcinoma was detected by cloning formation experiment after interfering KIF14.Flow cytometry was used to detect the apoptosis of cisplatin resistant cells in endometrial carcinoma after interfering KIF14,and western-blot was used to detect the expression levels of proliferation,apoptosis,and drug resistant gene related proteins,such as ki-67,Bax,cleaved Caspase-3,MDR1,and P-gp.The cells in each group were added to cisplatin with series of concentrations,and the sensitivity of cisplatin of cisplatin resistant cells in endometrial carcinoma was detected by MTT assay after interfering KIF14.Results:KIF14 was expressed highly in cisplatin-resistant endometrial cancer cell line Ishikawa/DDP compared with Ishikawa(P<0.001).Compared with those of the cells in drug-resistant group and si-NC group,the number of Ishikawa/DDP cell clone formation and ki-67 protein level in si-KIF14 group had decreased,and the cell apoptosis rate and the levels of Bax and Cleaved caspase-3 protein had increased,but the IC50 value had decreased after interfering KIF14.The levels of MDR1 and P-GP protein of the cells in si-KIF14 group had decreased(all P<0.001).Conclusion:Interfering KIF14 expression can inhibit the proliferation of Ishikawa/DDP of the endometrial carcinoma cells with platin resistant,which can promote apoptosis of the cells and increase their sensitivity to cisplatin.