基础医学与临床 ›› 2017, Vol. 37 ›› Issue (7): 1031-1036.

• 研究论文 • 上一篇    下一篇

下调HIPK2基因表达促进顺铂诱导的肾小管上皮HKC细胞凋亡

李佳垚,吴洁,邱玲   

  1. 北京协和医院
  • 收稿日期:2017-03-22 修回日期:2017-04-24 出版日期:2017-07-05 发布日期:2017-06-23
  • 通讯作者: 邱玲 E-mail:lingqiubj@aliyun.com

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

  • Received:2017-03-22 Revised:2017-04-24 Online:2017-07-05 Published:2017-06-23

摘要: 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,AnnexinV/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);转染小干扰RNA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。

关键词: HIPK2, HKC, 凋亡, 顺铂

Abstract: 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 real-time PCR and western blot. Two HIPK2 siRNAs were designed according to gene sequence of HIPK2 and cell lines with HIPK2 knockdownwere established through transfecting the HIPK2 siRNAs into HKC cells by liposome. The expression of HIPK2 mRNA and protein were detected by real-time PCR and western blot after induced by cisplatin. Then cell apoptosis was detected by Annexin V/PI after the HIPK2-knockdown cells were treated with cisplatin. Moreover, the expression of pro-apoptotic protein bax was detected by western blot after HIPK2 was knockdown. Results The expression level of HIPK2 mRNA and protein were down-regulated obviously on the process ofHKC apoptosis whichinduced by dose-dependent cisplatin (p<0.05). The transfection of siRNA can significantly reduce the expression of HIPK2 mRNA and protein in HKC (p<0.05), which will promote the HKC cells apoptosis induced by cisplatin. Conclusions HIPK2 can suppress the HKC cells apoptosis induced by cisplatin.

Key words: HIPK2, HKC, apoptosis, cisplatin

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