c-af-1反义寡核苷酸转染对人卵巢癌SKOV_3细胞增殖的抑制作用

Inhibitory effects of c-raf-1 antisense oligodeoxynucleotides transfactionon proliferation of the human ovarian carcinoma SKOV_3 cell lines

目的:探讨c-raf-1基因反义寡核苷酸转染对人卵巢癌SKOV3细胞增殖的抑制作用及其分子机制。方法:实验分3组:正常对照组、c-raf-1正义治疗组、c-raf-1反义治疗组。脂质体转染后不同时间分别进行M TT试验、集落形成试验、荧光显微镜检测、蛋白质荧光强度测定,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:反义治疗组与正义治疗组比较:转染72h OD值分别0.272与1.307(P<0.05);克隆形成率分别为30.6%与75.0%(P<0.05);凋亡率分别为19.7%与9.2%(P<0.05),同时明显地下调P74蛋白质的表达水平(P<0.05)。结论:c-raf-1反义寡核苷酸对人卵巢癌SKOV3细胞的增殖有明显的抑制作用。

Objective： To investigate the effects and the mechanisms of c-raf-1 genes antisense oligodeoxynucleotides （ ASODN ） transfection in inhibiting the human ovarian carcinoma SKOV3 cell lines. Methods： There were 3 groups in our study： normal control group, c-raf-1 sense oligodeoxynucleotides （ SODN ） experimental group, and c-raf-1 antisense experimental group, at the different time points after liposome-mediated transfection, the cell proliferation, apoptosis, protein expressing level were observed by MTT assay, flow cytometry, fluorescent microscope and cloning test. Results： In the ASODN experimental group and SODN group, the OD-value were 0.272 and 1.307 respectively （ P〈0.05 ） 72h after transfection; the cloning efficiency were 30.6% and 75.0% respectively （ P〈0.05 ） ; the apoptosis rate were 19.7% and 9.2% respectively （ P〈0.05 ） and the level of P74 protein was decreased statistically in c-raf-1 ASODN group. Conclusions： The proliferation of the human ovarian carcinoma SKOV3 cell lines can be inhibited by the c-raf-1 ASODN.