自然科学版
陕西师范大学学报(自然科学版)
生命科学
FAK表达水平对人肝癌细胞运动、增殖、凋亡等的影响
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张楠楠,魏丽丽,强荣兵,聂红娟,高晓杰,侯颖春*
(陕西师范大学 生命科学学院, 陕西 西安710062)
张楠楠,女,硕士研究生,主要研究方向为肿瘤相关基因功能.E-mail:zhangnannan@stu.snnu.edu.cn.* 通信作者:侯颖春,男,教授,博士研究生导师.E-mail:ychhou@snnu.edu.cn.
摘要:
构建了FAK基因siRNA质粒表达载体并用磷酸钙法转染人肝癌细胞SMMC—7721,采用半定量RT-PCR、Western Blot检测FAK siRNA转染后SMMC—7721细胞FAK mRNA和蛋白表达的变化.以损伤修复法检测FAK siRNA对SMMC—7721细胞迁移的影响,以四甲基偶氮唑盐 (MTT) 法检测 FAK siRNA转染对SMMC—7721细胞生长的抑制作用,以Hoechst染色法检测FAK siRNA对SMMC—7721细胞凋亡的影响.结果显示,特异性的FAK siRNA片段能有效降低SMMC—7721细胞FAK mRNA和蛋白的表达水平;转染FAK siRNA后,SMMC—7721细胞生长和迁移均受到抑制,凋亡细胞的数目也显著增多.表明靶向FAK基因的siRNA可以明显抑制SMMC—7721细胞的运动和增殖等生物学行为.
关键词:
粘膜斑激酶; RNAi; 肝癌; 细胞运动; 细胞增殖和凋亡
收稿日期:
2011-05-19
中图分类号:
R730.231
文献标识码:
A
文章编号:
1672-4291(2011)06-0072-04
基金项目:
陕西省自然科学基础研究计划项目(SJ08-ZT09);西安市科技局社会发展研发项目(YF07188).
Doi:
Effect of FAK expression level on the motility, proliferation and apoptosis of SMMC—7721 cells
ZHANG Nan-nan, WEI Li-li, QIANG Rong-bing, NIE Hong-juan, GAO Xiao-jie, HOU Ying-chun*
(College of Life Science, Shaanxi Normal University, Xi′an 710062, Shaanxi, China)
Abstract:
The siRNAs targeting FAK were designed and transfected into SMMC—7721 cells using a calcium phosphate transfection method. The changes of FAK expression level were detected with RT-PCR and western blot. The effects of FAK gene knockdown on migration, proliferation and apoptosis were investigated. The results show that FAK mRNA and protein level were significantly regulated down post the cells transfection by FAK siRNA. The proliferation and migration of SMMC—7721 cells were significantly inhibited, and the apoptosis of transfected cells were significantly enhanced. The results above indicate that the specific FAK siRNA can inhibits the motility and proliferation of SMMC—7721 cells.
KeyWords:
focal adhesion kinase (FAK); RNAi; hepatocarcinoma; cell motility; cell proliferation and apoptosis