摘要
目的研究靶向HER2/neu基因的siRNA对膀胱癌BIU-87细胞增殖和凋亡的影响。方法化学合成的靶向HER2/neu基因siRNA在脂质体的介导下转染BIU-87细胞,实验分为HER2/neu siRNA组、空脂质体组、阴性siRNA序列组,以未转染BIU-87细胞为空白对照组。利用CCK-8法评价HER2/neu基因siRNA对BIU-87细胞体外生长的抑制作用,流式细胞术检测细胞凋亡情况,通过逆转录聚合酶链反应(RT-PCR)和Western blot法检测转染前后细胞中HER2/neu mRNA和蛋白表达水平的变化。结果转染HER2/neu siRNA后BIU-87细胞的存活率显著下降,从(82.37±0.90)%降低到(56.76±1.70)%,差异有统计学意义(P<0.05);同时能诱导细胞凋亡,HER2/neu siRNA组凋亡率达(45.60±0.70)%,与空白对照组、空脂质体组、阴性siRNA序列组比较差异有统计学意义(P<0.05);靶向HER2/neu基因siRNA能显著降低BIU-87细胞中HER2/neu mRNA和蛋白的表达(P<0.05)。结论化学合成的靶向HER2/neu基因siRNA能有效抑制HER2/neu基因在BIU-87细胞中的表达,进而能有效抑制BIU-87细胞的增殖和诱导凋亡的作用。
Objective To investigate the effect of small interfering RNA （siRNA）-mediated gene silencing of HER2/neu on the proliferation and apoptosis in bladder cancer cell line BIU-87. Methods HER2/neu siRNA was chemically synthesized and transfected into BIU-87 cells mediated by liposome. BIU-87 cells with different treatments were divided into an HER2/neu siRNA group, an empty liposome group, a nega- tive siRNA group and a non-transfection group was the control group. Inhibitory effect of HER2/neu siRNA on BIU-87 cell proliferation was assessed by CCK-8 assay. Cell apoptosis was examined by flow cytometry （FCM）. mRNA and protein levels of HER2/neu in BIU-87 cells before and after transfection were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting. Results BIU-87 cells transfeeted with HER2/neu siRNA showed a significant decrease of cell viability from （ 82.37 _＋ 0.90 ） % to （ 56.76 _＋ 1.70） % （ P 〈 0.05 ） and cell apoptosis with a rate of （45.60 __. 0.70） %, which was significantly different from that of the control group, empty liposome group and negative HER2/neu siRNA group （ P 〈 0.05 ）. The siRNA targeting HER2/neu gene significantly reduced mRNA and the protein expressions of HER2/neu in BIU-87 cells （P 〈 0.05 ）. Conclusion Chemically synthesized siRNA targeting HER2/neu gene, which can effectively inhibit HER2/neu gene expression and subsequently inhibit cell proliferation and induce apoptosis in BIU-87 cells, is a potential target for gene therapy of bladder cancer.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第1期20-23,共4页
Journal of Third Military Medical University
基金
福建省科技厅青年人才项目(2007F3036)~~
