RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗敏感度的影响

目的 探讨RNA干扰沉默MSI1表达对人脑胶质瘤细胞化疗药物敏感度的影响。方法 构建针对MSI1 mRNA的小分子RNA干扰（siRNA）重组质粒pSIREN-MSI1，转染人脑胶质瘤U-87MG细胞；Western blot检测U-87MG细胞内MSI1蛋白的表达；CCK-8法观察U-87MG细胞对化疗药物替莫唑胺（TMZ）敏感度的改变；流式细胞仪（FCM）检测pSIREN-MSI1转染后对U-87MG细胞凋亡的影响。结果 成功构建了重组质粒pSIREN-MSI1，并成功转染U-87MG细胞；Western blot结果显示pSIREN-MSI1抑制U-87MG细胞中MSI1蛋白的表达（P<0.01）；CCK-8结果显示在替莫唑胺作用下，pSIREN-MSI1组U-87MG细胞的存活率明显下降（P<0.01）；FCM结果表明pSIREN-MSI1转染后明显提高U-87MG细胞的凋亡率（P<0.01）。结论 MSI1 siRNA能特异性抑制人脑胶质瘤细胞U-87MG细胞中MSI1的表达，增强人脑胶质瘤U-87MG细胞对化疗药物的敏感度，并促进细胞的凋亡。

Effect of RNA Interference Decreasing MSI1 Expression on Chemosensitivity of Human Glioma Cells

Objective To explore the effect of RNA interference decreasing MSI1 gene expression on the chemosensitivity of human glioma cells U-87MG. Methods The design and synthesis of Msi1- specific siRNA and the transfection of U-87MG cells were conducted. The expression of MSI1 in U-87MG cells was detected by Western blot. CCK-8 was employed to observe the sensitivity of U-87MG cells to temozolomide(TMZ). FCM was used to observe the apoptosis rate of U-87MG cells after pSIRENMSI1 transfection. Results Lentiviral vector-mediated MSI1-siRNA was successfully constructed. Immunofluorescence assay demonstrated that the transfection efficiency was above 60%. Western blot analyses demonstrated that pSIREN-MSI1 could significantly inhibit the expression of MSI1 in U-87MG cells (P<0.01); CCK-8 results showed that when U-87MG cells were exposed to temozolomide, the growth inhibiting rate of pSIREN-MSI1 transfected cells was significantly increased, compared with those of blank control group and negative control group (P<0.01). FCM results showed that the apoptosis rate of U-87MG cells were increased after pSIREN-MSI1 transfection(P<0.01). Conclusion MSI1-siRNA could significantly inhibit the expression of MSI1 in human glioma cells U-87MG. It could induce the apoptosis and enhance the chemosensitivity of U-87MG cells.