siRNA干扰叉头框C2对乳腺癌中肿瘤干细胞标志物CD44的影响  

Effect of forkhead box C2 gene silencing by siRNA on cancer stem cell marker CD44 in breast cancer

作　　者：王健[1] 徐杰[1] 吕健东[1] 

机构地区：[1]天津市第五中心医院普通外科,天津300450

出　　处：《中国普通外科杂志》2013年第11期1461-1465,共5页China Journal of General Surgery

摘　　要：目的:探讨siRNA干扰叉头框C2(FOXC2)对乳腺癌中肿瘤干细胞(CSC)标志物CD44 mRNA及蛋白表达的影响。方法:常规培养乳腺癌MCF-7细胞株,再通过乳腺球形成实验培养、筛选乳腺癌CSC;将FOXC2-siRNA或阴性对照siRNA慢病毒载体转染至乳腺癌CSC,同时将转染载体的CSC作为空白对照。通过real time RT-PCR和Western blot检测FOXC2-siRNA对FOXC2的干扰效应,再通过real time RT-PCR和Western blot分别检测CD44 mRNA和蛋白在各组细胞中的表达,结果均以空白对照的表达量作为参照进行分析。结果:从MCF-7细胞株中成功培养出乳腺癌CSC。与转染阴性对照siRNA的乳腺癌CSC比较,转染FOXC2-siRNA的乳腺癌CSC中FOXC2 mRNA和蛋白表达均明显降低(P=0.00509;P=0.00001),同时CD44 mRNA及蛋白的表达也均明显降低(P=0.00848;P=0.00218)。结论:干扰乳腺癌CSC中FOXC2基因的表达能够抑制乳腺癌CSC中CD44 mRNA及蛋白的表达,FOXC2信号转导通路可能通过调控CD44来介导乳腺癌CSC的增殖、分化。Objective： To investigate the effect of forkhead box C2 （FOXC2） gene silencing by siRNA on mRNA and protein expression of cancer stem cell （CSC） marker CD44 in breast cancer. Methods： Breast cancer MCF-7 cells were firstly cultured by routine methods, and then were cultured in mammosphere-forming conditions to enrich and screen out breast cancer CSCs. Next, the obtained CSCs were transfected with the lentiviral vector containing FOXC2-siRNA or negative control-siRNA, and CSCs transfected with empty vector were served as blank control. The FOXC2 mRNA and protein expressions in breast cancer CSCs were determined by real time RT-PCR and Western blot method, respectively, to assess the silencing efficacy, and then CD44 mRNA and protein expressions in each group of breast cancer CSCs were measured by real time RT-PCR and Western blot, respectively. All results were analyzed by comparing to the expression levels of blank control. Results： ~ae breast cancer CSCs were successfully cultured from MCF-7 cells by mammosphere formation. Compared with breast cancer CSCs transfected with negative control siRNA, both FOXC2 mRNA and protein expressions were decreased significantly （P=0.00S09, P=O.O0001）, and both CD44 mRNA and protein levels were down-regulated significantly in FOXC2-siRNA transfected breast cancer CSCs （P=0.00848, P=0.00218）. Conclusion： FOXC2 gene silencing can inhibit the CD44 mRNA and protein expression in breast cancer CSCs. So, FOXC2 signaling pathway may possibly participate in the proliferation and differentiation process of breast cancer CSCs through regulating CD44 expression.

关 键 词：乳腺肿瘤 肿瘤干细胞 叉头框C2 抗原 CD44 

分 类 号：R737.9[医药卫生—肿瘤]