Zhang Shu-Cheng, Qiao Lei, Gao Hong
Department of Pediatric Surgery, Shengjing Hospital, China Medical University, Shenyang 110004, China.
Zhonghua Wai Ke Za Zhi. 2007 Apr 1;45(7):473-5.
To explore whether the inducible knockdown of endogenous CXCR4 gene expression in breast cancer cells can inhibit the migration significantly in vitro.
The hairpin siRNA templates were designed and the siRNA expression cassettes (SECs) were generated. In vitro the SECs were transfected into MDA-MB-231 cells using the agent siPORTTM XP-1. The non-transfected cells were taken as control. The inhibitory effect of siRNAs was detected by RT-PCR and Western blot. The invasion ability of breast cancer cell was assayed in 24-well Biocoat Matrigel invasion chambers.
The endogenous CXCR4 gene expression could be specifically knocked down by the SECs detected with both RT-PCR and Western blot. After the inducible knockdown, the invasion of the breast cancer cell in CXCR4 group was significantly inhibited compared with that of controls (P < 0.05)
It is showed that inducible knockdown of endogenous CXCR4 gene expression in breast cancer cells can result in significant inhibition of breast cancer cell migration in vitro. Overexpression of CXCR4 is related with the migration of breast cancer cells.
探讨乳腺癌细胞内源性CXCR4基因表达的诱导性敲低是否能在体外显著抑制其迁移。
设计发夹状siRNA模板并构建siRNA表达盒（SECs）。体外使用siPORTTM XP-1试剂将SECs转染至MDA-MB-231细胞中，未转染的细胞作为对照。通过RT-PCR和蛋白质印迹法检测siRNAs的抑制效果。在24孔BioCoat Matrigel侵袭小室中检测乳腺癌细胞的侵袭能力。
RT-PCR和蛋白质印迹法检测均显示SECs可特异性敲低内源性CXCR4基因表达。诱导性敲低后，CXCR4组乳腺癌细胞的侵袭与对照组相比显著受到抑制（P < 0.05）。
结果表明，乳腺癌细胞内源性CXCR4基因表达的诱导性敲低可导致体外乳腺癌细胞迁移受到显著抑制。CXCR4的过表达与乳腺癌细胞的迁移有关。
