靶向Her2基因SiRNA对人卵巢癌细胞株SKOV-3顺铂敏感性的影响  

The effects of siRNA specific for Her-2 gene on the cisplatin sensitivity of ovarian carcinoma cell line

作　　者：程志祥[1] 周颖[1] 朱园园[1] 肖敏[1] 高宗侠[1] 祝怀平[1] 凌斌[1] 

机构地区：[1]安徽医科大学附属省立医院分子实验室,合肥230001

出　　处：《安徽医科大学学报》2007年第4期374-378,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金项目(编号:20050430715);安徽省教育厅自然科学研究项目(编号:2005KJ347zc)

摘　　要：目的通过siRNA沉默Her2基因的表达探讨其对卵巢癌细胞株SKOV-3顺铂(DDP)敏感性的影响。方法靶向Her2的siRNA通过阳离子脂质体介导转染到SKOV-3中,经过嘌呤霉素筛选得到稳定抑制Her2基因表达的SK-OV-3细胞株,通过RT-PCR方法检测各组SKOV-3细胞Her2基因的表达。应用四甲基偶氮唑蓝检测DDP对各组SKOV-3细胞增殖水平的影响,应用膜联蛋白V异-硫氰酸荧光素细胞凋亡试剂盒在流式细胞仪上检测DDP作用后各组SKOV-3细胞凋亡水平。结果建立了稳定抑制Her2基因表达的细胞株,SKOV3/siRNA-1组和SKOV3/siRNA-2组细胞Her2基因表达分别为对照组的53.8%和38.4%,而空载体组为92.3%。在DDP作用下SKOV3/siRNA-1和SKOV3/siRNA-2的细胞存活率在4 d时分别为67.7%±4.4%和66.4%±3.3%,与未转染对照组81.7%±1.3%和空载体组81.3%±1.4%比较差异有显著性(P<0.05)。FCM分析显示SK-OV3/siRNA-1和SKOV3/siRNA-2细胞的DDP诱导凋亡率分别为38.6%±1.1%和42.8%±1.4%,明显高于未转染组9.1%±0.8%和空载体组8.8%±0.7%,差异有显著性(P<0.05)。结论靶向Her2基因的siRNA可以增强卵巢癌细胞株SKOV-3对DDP的敏感性。Objective To explore the drug sensitivity of ovarian carcinoma cell line SKOV-3 to cisplatin（DDP） by siRNA specific for Her2 gene. Methods The experiments consisted of four groups including an untreated control group, an empty vector group, a SKOV-3/siRNA-1 group, a SKOV-3/siRNA-2 group. The two specific vectorsbased siRNAs were transfected into cell line SKOV-3 by Lipofectamine 2000 individually, and two kinds of stable cell line with a persistent silencing of Her2 gene were established by selecting with puromycin. Significantly reducing of Her2 mRNA expressed in those cells showed by RT-PCR. The chemosensitivity of transfected cells to DDP was measured by MTT. Cell apoptosis detection kit （AnnexinV-FITC） was used to examine the drug induced apoptosis rate. Results The stable SKOV-3 cell lines with a persistent knockdown of Her2 were established, the Her2 gene expression rate in the SKOV-3/siRNA-1 group, the SKOV-3/siRNA-2 group and the empty vector group were 53.8% ,38.4% ,92. 3%, respectively. Consequently the survival rate of siRNA-1 group affected by DDP at 4th day was 67.7% ±4. 4% and siRNA-2 group was 66.4%± 3.3%, indicated significant difference between untreated control group [ （81.7% ± 1.3% ） ] or empty vector group （81.3% ±1.4% ,P 〈0. 05）. FCM results showed the apoptosis rates of the SKOV3/siRNA-1 and 2 groups induced by DDP were increased to 38. 6% ± 1.1% and 42. 8% ± 1.4%, respectively. There was significant difference between siRNA groups and the other groups （ P 〈 0. 05）. Conclusion The sensitivity to DDP can be increased by inhibition the expression of Her-2 gene.

关 键 词：基因 ERBB-2 RNA 小分子干扰 顺铂 卵巢肿瘤 

分 类 号：R737.31[医药卫生—肿瘤] R979.19[医药卫生—临床医学]