摘要
目的探讨抑制趋化因子受体 CXCR4基因对乳腺癌细胞转移和侵袭能力的影响。方法将人乳腺癌细胞株 MDA-MB-231分为未干扰组、CXCR4干扰组、β-actin 干扰组和空白载体干扰组。设计并合成了针对 CXCR4基因的发夹式 siRNA 模板,体外合成 siRNA 表达框架(siRNAexpression cassettes,SECs),经脂质体转染 CXCR4干扰组细胞,采用 Western blot 免疫印迹法和逆转录聚合酶链反应检测各组的抑制效果,用 Biocoat Matrigel 侵袭能力检测仪检测 MDA-MB-231细胞的转移侵袭能力。结果 SECs 在 mRNA 水平和蛋白水平均明显抑制 CXCR4基因的表达。CXCR4表达被抑制后,癌细胞侵袭能力受到明显抑制,与未干扰组相比,差异有统计学意义(P<0.05)。结论SECs 可以高效特异地抑制人乳腺癌细胞中 CXCR4基因的表达。CXCR4基因高表达与乳腺癌细胞的转移密切相关,抑制细胞中 CXCR4的表达可以抑制乳腺癌细胞转移和侵袭能力。
Objective To explore whether the inducible knockdown of endogenous CXCR4 gene expression in breast cancer cells can inhibit the migration significantly in vitro. Methods The hairpin siRNA templates were designed and the siRNA expression cassettes （SECs） were generated. In vitro the SECs were transfected into MDA-MB-231 ceils using the agent siPO2RTrMXP21. The non-transfected ceils were taken as controL The inhibitory effect of siRNAs was detected by RT-PCR and Western blot. The invasion ability of breast cancer ceil was assayed in 24-well Biocoat Matrigel invasion chambers. Results 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 ceil in CXCR4 group was significantly inhibited compared with that of controls （ P 〈 0. 05 ） Conclusions It is showed that inducible knockdown of endogenous CXCR4 gene expression in breast cancer ceils can result in significant inhibition of breast cancer ceil migration in vitro. Overexpression of CXCR4 is raleted with the migration of breast cancer cells.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2007年第7期473-475,共3页
Chinese Journal of Surgery
