摘要
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p SIREN-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p SIREN-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p SIREN-MSI1,并成功转染U-87MG细胞;Western blot结果显示p SIREN-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 si RNA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。
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.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2016年第9期754-757,共4页
Cancer Research on Prevention and Treatment
作者简介
作者简介：刘细国（1980-），男，博士，主治医师，主要从事脑肿瘤的基础与I临床研究
通信作者：付锴，E-mail：kaizige@hotmail．com
