抑制RhoA基因表达诱导乳腺癌细胞凋亡的实验研究

Inhibiting RhoA expression to induce apoptosis of breast cancer cells

目的探讨抑制RhoA基因表达诱导乳腺癌(BC)细胞凋亡的作用。方法通过已建立的RhoA腺病毒载体系统,感染BC MCF-7细胞。经过细胞培养、病毒滴度等,将收集的样本分别用Western blot、RT-PCR技术检测RhoA蛋白表达和基因表达变化;对感染Adsi RNA-RhoA的BC细胞进行MTT检测及细胞内DNA片段化检测。结果腺病毒siRNA-RhoA感染BC细胞能明显抑制RhoA基因表达(抑制率为75.64%),降低RhoA基因的mRNA转录水平69.44%。MTT检测结果表明,感染腺病毒Adsi RNA-RhoA组的肿瘤细胞生长、增殖被明显抑制。TUNEL检测显示,抑制RhoA基因表达能明显导致BC细胞凋亡。结论利用siRNA抑制BC细胞中RhoA基因表达,能诱导BC细胞凋亡。

Objective To investigate the role of RhoA gene inhibition in inducing the apoptosis of breast cancer （BC） cells.Methods Based on a previously constructed adenoviral vector-mediated siRNA system, we infected the BC MCF-7 cells.After cell culture and virus titer measurement experiment , we collected the virus sample .Then Western blot -ting and RT-PCR were used to detect the expression levels of protein and mRNA , respectively .MCF-7 cells were infected by Adsi RNA-RhoA and detected by using MTT and DNA fragmentation assays .Results The Adsi RNA-RhoA infected BC cells significantly inhibited the expression of RhoA gene （ the inhibition rate 75.64%） and reduced the transcriptional level of RhoA mRNA （69.44%）.MTT assay revealed that the growth and proliferation of MCF-7 cells infected by Adsi RNA-RhoA was inhibited .TUNEL assay indicated that Adsi RNA-RhoA could induce the apoptosis of MCF-7 cells.Con-clusion Using siRNA to inhibit the expression of RhoA gene in BC cells can induce the apoptosis of BC cells .