下调HIPK2基因表达促进顺铂诱导的肾小管上皮细胞HKC凋亡

Down-regulating HIPK2 promotes cisplatin-induced apoptosis of human kidney tubular epithelial cells

目的探讨HIPK2对顺铂诱导的人肾小管上皮细胞(HKC)凋亡的影响。方法构建顺铂诱导的HKC细胞凋亡模型,实时荧光定量PCR和Western blot检测HIPK2的表达;设计合成2条HIPK2 siRNA干扰片段,通过脂质体转染HKC细胞,建立HIPK2干扰的细胞株;实时荧光定量PCR和Western blot分别检测HIPK2 mRNA和蛋白的表达;再用顺铂处理细胞,Annexin V/PI检测细胞凋亡;Western blot检测Bax表达的影响。结果顺铂呈剂量依赖性诱导的HKC细胞凋亡过程中,HIPK2 mRNA和蛋白表达水平均明显下调(P<0.05);转染siRNA后可显著降低HIPK2在HKC细胞内mRNA和蛋白的表达(P<0.05),且HIPK2表达降低后会促进顺铂诱导的HKC细胞凋亡。结论HIPK2可抑制顺铂诱导的HKC细胞凋亡。

Objective To explore the effect of HIPK2 on apoptosis of human kidney tubular epithelial cells （HKC） induced by cisplatin. Methods Apoptosis of HKC cells was induced by cisplatin and the expression of HIPK2 was detected by RT-qPCR and Western blot. Two HIPK2 siRNAs were designed according to gene sequence of HIPK2 and cell lines with HIPK2 knockdown were established through transfecting the HIPK2 siRNAs into HKC cells by lipo- some. The expression of HIPK2 mRNA and protein was detected by RT-qPCR and Western blot after induced by cis- platin. Then cell apoptosis was detected by Annexin V/PI after the HIPK2-knockdown cells were treated with cispla- tin. Moreover, the expression of pro-apoptotic protein bax was detected by Western blot after HIPK2 was knockdown. Results The expression of HIPK2 mRNA and protein was down-regulated obviously on the process of HKC apoptosis which induced by dose-dependent cisplatin （P〈0. 05）. The transfection of siRNA could significantly reduce the ex- pression of HIPK2 mRNA and protein in HKC （P〈0. 05）, which promotes the HKC cells apoptosis induced by cispl- atin. Conclusions HIPK2 can suppress the HKC cells apoptosis induced by cisplatin.