基础医学与临床 ›› 2018, Vol. 38 ›› Issue (1): 74-79.

• 研究论文 • 上一篇    下一篇

RNA干扰下调RNF2抑制胰腺癌细胞系PANC-1增殖和迁移

吴银芳1,2,孙晓东1,郑宇1,黄东胜1   

  1. 1. 浙江省人民医院
    2. 浙江中医药大学
  • 收稿日期:2017-09-14 修回日期:2017-11-18 出版日期:2018-01-05 发布日期:2017-12-26
  • 通讯作者: 孙晓东 E-mail:sunxiaodong@hmc.edu.cn
  • 基金资助:
    浙江省自然科学基金项目

RNA interference targeting RNF2 inhibits proliferation and migration of pancreatic cancer cell line PANC-1

  • Received:2017-09-14 Revised:2017-11-18 Online:2018-01-05 Published:2017-12-26
  • Contact: Xiao-Dong Sun E-mail:sunxiaodong@hmc.edu.cn
  • Supported by:
    Zhejiang Provincial Natural Science Foundation of China

摘要: 目的 研究沉默环指蛋白2(RNF2)基因对人胰腺癌PANC-1细胞增殖、迁移、周期和凋亡的影响及可能机制。方法 用siRNA-RNF2转染PANC-1细胞沉默RNF2表达,同时设立转染无义序列(siRNA-NC)的空转染组及不进行任何处理的空白对照组(mock)。荧光定量PCR检测RNF2 mRNA表达。Western blot检测RNF2和p53表达。MTS实验和划痕实验分别检测细胞增殖和迁移能力。流式细胞计量术检测细胞转染率、凋亡率和细胞周期。结果 相比正常胰腺导管上皮细胞,RNF2在胰腺癌细胞系中高表达(P<0.05)。转染siRNA-RNF2的PANC-1细胞与阴性对照组比较,RNF2 mRNA及蛋白表达下调;细胞增殖被抑制(P<0.05);细胞迁移能力下降(P<0.05);细胞凋亡率增高(P<0.05)和G0/G1期细胞比例上升、S期和G2/M期细胞比例则降低(P<0.05)。此外,转染siRNA-RNF2显著下调PANC-1细胞中p53蛋白的表达。结论 siRNA-RNF2特异性下调胰腺癌细胞RNF2表达,显著抑制细胞增殖和迁移,提示RNF2可能成为胰腺癌基因治疗的一个新靶点。

关键词: 环指蛋白2, 胰腺癌, 生长抑制, p53

Abstract: Objective To investigate the effect of siRNA-mediated silencing of RNF2 gene on cell proliferation, migration, cell cycle and apoptosis in human pancreatic cancer PANC-1 cells and its possible mechanism. Methods The siRNA interference was used to down-regulate RNF2 expression. Meanwhile, there were also empty transfection group whose cells were transfected with the control siRNA and mock group without any treatment. The result of transfection was evaluated by fluorescence microscope. The expression of RNF2 mRNA was detected by RT-qPCR. Western blot was applied to detect the expression of RNF2 and p53. Cell proliferation and migration were analyzed by MTS assay and cell scratch assay, respectively. The transient transfection efficiency, apoptosis rate and cell cycle were measured by flow cytometry. Results Compared to the normalized human pancreatic duct epithelial cells, RNF2 expression in pancreatic cancer cells were higher (P<0.05). The expression of RNF2 mRNA and protein was decreased in PANC-1 cells by siRNA-RNF2 at 48 h post-transfection. Transfection with siRNA-RNF2 inhibited the proliferation and migration of PANC-1 cells (P<0.05), induced cell apoptosis (P<0.05), increased cell counts in phase G0/G1 and decreased in S and G2/M phase (P<0.05). What’s more, after siRNA-RNF2 transfection, the expression of p53 protein was decreased. Conclusions siRNA-RNF2 can specifically knockdown the expression of RNF2 gene and inhibit the proliferation and migration of PANC-1 cells. These results indicate RNF2 may thus become a new target of gene therapy for pancreatic cancer.

Key words: RING finger protein 2, pancreatic cancer, growth inhibition, p53

中图分类号: