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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 FOXC2 knockdown on CD44 mRNA and protein expression of cancer stem cell (CSC) in breast cancer. Methods: Breast cancer MCF-7 cells were cultured routinely and then cultured for CSC screening. The FOXC2-siRNA or negative control siRNA lentiviral vector was transfected into breast cancer CSC, and the CSC As a blank control. The effect of FOXC2-siRNA on FOXC2 was detected by real time RT-PCR and Western blot. The expression of CD44 mRNA and protein in each group was detected by real time RT-PCR and Western blot respectively. The results were expressed as blank control The amount is analyzed as a reference. Results: Breast cancer CSC was successfully cultured from MCF-7 cell line. Compared with breast cancer CSC transfected with negative control siRNA, FOXC2 mRNA and protein expression in breast cancer CSCs transfected with FOXC2-siRNA were significantly decreased (P = 0.00509; P = 0.00001), meanwhile CD44 mRNA and protein expression were also significantly lower Decreased (P = 0.00848; P = 0.00218). CONCLUSION: The FOXC2 gene expression in breast cancer CSC can inhibit the expression of CD44 mRNA and protein in breast CSC. FOXC2 signal transduction pathway may mediate the proliferation and differentiation of CSC in breast cancer by regulating CD44.