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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-time PCR和Western blot方法,分别从mRNA水平及蛋白水平检测IGF-IR的表达;通过MTT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pSih IGF-IR-1、pSih IGF-IR-2和pSih IGF-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一条新的途径。
Objective To investigate the significance of silencing IGF-IR in RNA interference targeted therapy of ovarian cancer. Methods The recombinant plasmid was designed and synthesized based on IGF-IR gene. The expression of IGF-IR was detected by Real-time PCR and Western blot respectively. The cell proliferation was detected by MTT assay. RESULTS: Three expression vectors pSih IGF-IR-1, pSih IGF-IR-2 and pSih IGF-IR-3 were successfully constructed and transfected into ovarian cancer OVCAR3 cells. After 48 hours, the mRNA and protein levels of IGF- Significantly decreased, and significantly inhibited OVCAR3 cell proliferation in vitro. Conclusion The RNA interference technique can efficiently and specifically silence IGF-IR gene expression and inhibit tumor growth. RNAi for ovarian cancer IGF-IR technology may open a new way for the clinical treatment of ovarian cancer.