RNA干扰沉默IGF—IR基因治疗卵巢癌的实验研究

目的研究RNA干扰技术沉默IGF—IR在卵巢癌靶向治疗中的意义。方法以IGF—IR为目的基因,设计合成siRNA重组质粒;利用Real—timePCR和Westernblot方法,分别从mRNA水平及蛋白水平检测IGF—IR的表达;通过MTF实验检测细胞增殖情况。结果本实验成功构建了三个表达载体pSihIGF—IR-1、pSihIGF—IR-2和pSihIGF—IR-3,转染卵巢癌OVCAR3细胞,48h后,IGF—IR的mRNA及蛋白水平均明显降低,并且显著抑制了OVCAR3细胞的体外增殖。结论应用RNA干扰技术,能够高效、特异的沉默IGF—IR基因的表达,抑制肿瘤生长。针对卵巢癌IGF—IR的RNAi技术,可能为卵巢癌的临床治疗开拓-条新的途径。

Experimental study of insulin - like growth factor - I receptor silencing mediated by RNA interference in ovarian cancer therapy.

Objective ： To investigate the possibility of IGF - IR small interfering RNA （siRNA） as a promising anticancer agent. Methods： The technology of RNA interference was used to silence IGF - IR gene expression in the human ovarian cancer cell line OVCAR3. Real - time PCR and western blot assay were used to observe the mRNA and protein expression of IGF - IR, and then evaluate its effects on proliferation of tumor cells in vitro. Results ： Three siRNA sequences for the IGF - IR gene were cloned into expres- sion plasmids and transfected into OVCAR3 ceils. The mRNA and protein expression of IGF - IR were significantly decreased at 48h after transfection, leading to the potent suppression of tumor cell proliferation in vitro. Conclusions： IGF - IR - specific siRNA suppresses tumor growth, which could define a potential therapeutic target.