生物学杂志

生物学杂志 ›› 2022, Vol. 39 ›› Issue (6): 30-.doi: 10.3969/j.issn.2095-1736.2022.06.030

• 研究报告 • 上一篇    下一篇

干扰VDR对肝癌HepG2细胞增殖的作用

  

  1. 陕西理工大学 生物科学与工程学院， 汉中 723000
  • 出版日期:2022-12-18 发布日期:2022-12-12
  • 通讯作者: 张涛，博士，教授，研究方向为生化与分子生物学，E-mail:zl780823@163.com
  • 作者简介:程佳，博士，研究方向为细胞生物学，E-mail: chengjia0471@hotmail.com
  • 基金资助:
    陕西省科技厅项目（2018NY-157）； 陕西理工大学2020年大学生创新创业训练计划项目（109）

The effect of interfering VDR on HepG2 cell proliferation

  1. Biological Science ＆ Engineering Department, Shaanxi University of Technology, Hanzhong 723000, China
  • Online:2022-12-18 Published:2022-12-12

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摘要： 为研究维生素D受体（vitamin D receptor，VDR）对HepG2细胞增殖作用的调控，明确VDR对肝癌发生发展的意义。利用VDR基因特异性siRNA（small interfering RNA）转染HepG2细胞，实时荧光定量PCR(real-time quantitative PCR, qPCR)和蛋白印迹（western blot, WB）的检测结果发现，与对照NC siRNA相比，VDR siRNA-507显著抑制VDR的mRNA和蛋白表达水平（P<0.05）。CellTiter-Lumi发光法、EdU染色技术和细胞集落形成实验结果发现，干扰VDR对HepG2细胞增殖有极显著促进作用(P<0.01)。免疫荧光(immunofluorescence, IF)和双萤光素酶活性检测(double luciferase activity assay)分析结果显示，干扰VDR促进HepG2细胞增殖的过程与视黄醇结合蛋白4(retinol-binding protein, RBP4)的表达无关。

关键词: 维生素D受体, HepG2细胞, siRNA, 细胞增殖, 视黄醇结合蛋白4

Abstract: To investigate the effect of vitamin D receptor (VDR) in HepG2 cell proliferation and the significance of VDR in liver cancer, HepG2 cells were transfected with VDR gene-specific siRNA (small interfering RNA). The qPCR and WB results showed that compared with the control NC siRNA group, VDR siRNA-507 could significantly inhibit the mRNA and protein levels of VDR (P<0.05). The celltiter-Lumi luminescence, EDU staining and cell colony formation assay were employed to detect the effect of interference VDR on the proliferation of HepG2 cells. The results confirmed that interference with VDR could significantly promote the proliferation of HepG2 cells (P<0.01). The immunofluorescence and double luciferase activity assay were used to monitor the RBP4 expression during HepG2 cells growth process. However, interfering VDR had no effect on RBP4 expression level in HepG2. It was concluded that interference with VDR could promote HepG2 cells proliferation which was no relation with RBP4 level.

Key words: vitamin D receptor, HepG2 cell, small interfering RNA, cell multiplication, retinol binding protein 4

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