c-erbB-2与c-raf-1反义寡核苷酸转染对卵巢癌SKOV_3细胞的作用  

Effects of c-erbB-2 and c-raf-1 Antisense Oligodeoxynucleotides Transfected into the Human Ovarian Carcinoma SKOV3 Cell Line

作　　者：吴永忠[1] 任庆兰[1] 李少林[2] 

机构地区：[1]重庆医科大学附属第一医院肿瘤科,重庆市400016 [2]重庆医科大学基础医学院核医学教研室

出　　处：《中国肿瘤临床》2006年第14期784-787,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金资助(编号:30070230)

摘　　要：目的:探讨c-erbB-2与c-raf-1基因ASODN联合转染对人卵巢癌细胞增殖的抑制作用及其分子机制。方法:实验分为7组:对照组、c-erbB-2正义治疗组、c-raf-1正义治疗组、c-erbB-2反义治疗组、c-raf-1反义治疗组、全剂量联合治疗组和半剂量联合治疗组。脂质体转染后不同时间分别进行MTT试验、集落形成试验、荧光显微镜检测、蛋白质荧光强度测定与免疫组织化学检查,以观察不同条件处理后各组细胞的增殖、凋亡、蛋白质表达有无差别。结果:与对照组比较,全剂量联合治疗组和半剂量联合治疗组,转染72h的光密度值分别为0.151(P<0.005)和0.073(P<0.005),增殖抑制率分别达到了88.7%和94.5%,克隆形成率分别为19.4%(P<0.01)和12.8%(P<0.01),凋亡率分别为24.3%(P<0.05)和31.5%(P<0.01),同时明显地下调了c-erbB-2、c-raf-1、PCNA、c-jun、c-fos基因蛋白质的表达水平。结论:c-erbB-2与c-raf-1基因反义寡核苷酸联合转染对人卵巢癌SKOV3细胞增殖的抑制作用优于单反义基因,其对卵巢癌细胞增殖的抑制作用可能通过下调c-erbB-2、c-raf-1、PCNA、c-jun、c-fos基因蛋白的表达水平实现。Objective： To investigate the effects of c-erbB-2 and c-raf-1 ASODN transfected in combination into the human ovarian carcinoma SKOV3 cell line and the molecular mechanism involved. Methods： There were 7 groups in our study： control group, c-erbB-2 sense experimental group, c-raf- 1 sense experimental group, c-erbB-2 antisense experimental group, c-raf-1 antisense experimental group, whole-dose experimental group, and half-dose experimental group. At different timepoints after liposome-mediated transfection, the cell proliferation, apoptosis, and protein expression level were observed by MTY assay, flow cytometry, fluorescent microscopy, cloning test and immunohistochemistry. Results： Seventy-two hours after transfection the whole-dose experimental group and the half-dose experimental group had OD values of 0.151 （P〈0.005） and 0.073 （P〈0.005）, respectively. Compared to the control group, the whole-dose experimental group and the half-dose experimental group had proliferation rates that were 88.7% and 94.5%, respectively. The cloning efficiency was 19.4% （P〈0.01） and 12.8% （P〈0.01）, respectively, and the apoptosis rate was 24.3% （P〈0.05） and 31.5% （P〈0.01）, respectively. Protein levels including c-erbB-2, c-raf-1, PCNA, c-jun, and c-fos were downregulated. Conclusions： The study implies that c-erbB-2 and c-raf-1 combined ASODN transfection is superior to single ASODN for inhibiting proliferation of the human ovarian carcinoma SKOV3 cell line. It would be realized by regulating down the expression of c-erbB-2, c-raf-1, PCNA, c-jun, c-fos genes.

关 键 词：SKOV3细胞 反义寡核苷酸 联合转染 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]