中国临床解剖学杂志 ›› 2013, Vol. 31 ›› Issue (2): 170-173.

• 实验研究 • 上一篇    下一篇

siRNA沉默TAp73对骨肉瘤细胞的增殖和侵袭力的作用研究

薛剑, 靳安民, 吕海, 闵少雄   

  1. 南方医科大学珠江医院骨科,  广州   510282
  • 收稿日期:2012-10-25 发布日期:2013-04-08
  • 通讯作者: 靳安民,主任医师,博士生导师,E-mail:jinanmin@yahoo. com E-mail:gdwjxj@163.com
  • 作者简介:薛剑(1978-),男,广西梧州人,主治医师,博士,主要从事骨肿瘤方面的研究,Tel:(020)61627215
  • 基金资助:

    国家自然科学基金(81172555)

The influence of down-regulation of TAp73 expression by short hairpin RNA on the proliferation and invasion of osteosarcoma

XUE Jian, JIN An-min, LU Hai, MIN Shao-xiong   

  1. Department of Orthopaedic Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China
  • Received:2012-10-25 Published:2013-04-08

摘要:

目的 探讨通过siRNA沉默细胞TAp73蛋白表达对骨肉瘤Saos-2细胞的增殖、周期以及侵袭力的作用研究。  方法    以骨肉瘤Saos-2细胞系为研究对象,设计合成沉默TAp73的siRNA片段,转染入Saos-2细胞,用Western-blot法检测干扰后TAp73蛋白的表达水平,CCK-8法检测细胞增殖,利用流式细胞仪检测细胞周期,Transwell法检测细胞侵袭能力。  结果    Western-blot检测显示siRNA对TAp73蛋白表达有显著下调作用;CCK-8检测实验组细胞增殖明显(P<0.05);细胞周期检测显示细胞处于DNA合成期(S期)的细胞比例上升;穿过人工基底膜的细胞数量也明显增加,由(84.17±1.35)增加为(92.67±2.80 )(P<0.05)。  结论    抗癌基因TAp73在骨肉瘤的增殖、凋亡和侵袭过程中发挥有重要作用,可能成为控制肿瘤增殖和转移的关键性因素。

关键词: 骨肉瘤Saos-2细胞, TAp73, siRNA, 侵袭, 增殖

Abstract:

Objective To  investigate  the  pro-apoptosis,  anti-migration  and anti-proliferation of Saos-2 cells in osteosarcoma through genetic inhibition of expression of TAp73 protein. Methods Human osteosarcoma cell line Saos-2 was cultured.A short hairpin RNA targeting TAp73 was constructed and transfected into Saos-2 cell line.The expression of TAp73 protein were examined by Western blot method.The proliferation was assayed by CCK-8.Flow cytometry was used to detect the cell cycle.Transwell test was used to detect the invasion ability of Saos-2. Results Western blot revealed notably down-regulated expression at protein levels of Tap 73.The cellular proliferation in the treatment group detected by CCK-8 was active after siRNAtreatment(P<0.05).The proportion of the cells in the S phase increased.The invasion ability increased from( 84.17±1.35) to (92.67±2.80 )(P<0.01). Conclusions TAp73 is necessary for human osteosarcoma cell proliferation and invasion.It is probably a potential target for osteosareoma treatment.

Key words: Osteosarcoma, Tap73, SiRNA, Proliferation, Invasion

中图分类号: