解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (5 ): 656-660.doi: 10.3969/j.issn.0529-1356.2013.05.014

• 肿瘤生物学 • 上一篇    下一篇

膜联蛋白A7低表达对人肝癌HepG2细胞增殖的影响

王小杰  李欣*   

  1. 承德医学院基础医学研究所，河北 承德 067000
  • 收稿日期:2013-02-01 修回日期:2013-04-16 出版日期:2013-10-06 发布日期:2013-10-06
  • 通讯作者: 李欣 E-mail:cd-lixin@163.com

Influence of annexin A7 low expression on proliferation of HepG2 cells

WANG Xiao-jie LI Xin*   

  1. Basic Medical Institute, Chengde Medical University, Hebei Chengde 067000, China
  • Received:2013-02-01 Revised:2013-04-16 Online:2013-10-06 Published:2013-10-06

摘要：

目的 探讨膜联蛋白A7低表达对人肝癌HepG2细胞的增殖产生的影响。方法 采用Western blotting法鉴定siRNA可有效抑制膜联蛋白A7表达，然后用脂质体转染法将siRNA转染入HepG2细胞，将细胞分为siRNA干扰组、阴性对照组和空白对照组，在转染后24h、48h、72h进行细胞计数以绘制细胞增殖曲线，转染后48h进行MTT实验以检测细胞增殖活力；免疫组织化学法检测cyclinD1和Ki67的表达，Western blotting法检测cyclinD1的表达情况。结果 转染了靶向膜联蛋白A7的siRNA后48h的HepG2细胞，细胞计数和MTT实验可见siRNA干扰组细胞增殖活力较阴性对照组和空白对照组显著降低（P<0.05）；cyclinD1和Ki67蛋白表达均为siRNA干扰组细胞表达显著低于两对照组（P<0.05）。 结论 膜联蛋白A7低表达可能对HepG2细胞的增殖具有一定的抑制作用。

关键词:  膜联蛋白A7, HepG2细胞, cyclinD1, Ki67, 免疫组织化学, 免疫印迹法, 人

Abstract:

Objective To study whether annexin A7(ANXA7) low expression can affect proliferation of HepG2 cells. Methods Western blotting was used to identify siRNA can highly surpress annexin A7 expression. When the effect of the siRNA was assured, the siRNA was transfected into HepG2 cells by lipofectamine transfection method. At the same time, cells in negative control group were transfected negative siRNA while cells in blank control group only added lipofectamine 2000. Twenty-four, forty-eight and seventy-two hours after transfection ,cell counting was used to draw growth curve. Forty-eight hours after transfection, MTT assay was used to detect proliferation activity; Immunohistochemistry was applied to observe the expression of cyclinD1 and Ki67; Western blotting was performed to detect the expression of cyclinD1. Results Cell counting and MTT assay showed that proliferation of the siRNA interference group decreased significantly(P<0.05). The expression of cyclinD1 and Ki67 decreased compared with the negative control group and the blank control group(P<0.05). Conclusion ANXA7 low expression may inhibit proliferation of HepG2 cells.

Key words: ANXA7, HepG2 cell, cyclinD1, Ki67, Immunohistochemistry, Western blotting, Human