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目的:通过体内动物实验研究CD59-siRNA对卵巢癌移植瘤CD59的沉默效应及其抑瘤作用,探讨CD59在肿瘤免疫逃逸中的作用。方法:采用脂质体转染法将CD59干扰质粒(T组)和空质粒(V组)转染A2780细胞,获得稳定表达细胞株,将前两组A2780细胞和未转染(C组)的A2780细胞分别接种于裸鼠皮下建立肿瘤模型,通过绘制肿瘤生长曲线、RT-PCR和Western Blot研究其抑瘤效应及对CD59的沉默效应。结果:肿瘤生长曲线显示,与对照组相比,CD59干扰质粒转染组肿瘤生长明显受抑制(P<0.05)。RT-PCR和Western Blot结果表明,干扰组的CD59mRNA及CD59蛋白与对照组相比显著降低(P<0.05)。结论:动物实验表明,特异性沉默CD59基因的siRNA表达载体可以明显抑制CD59的表达及卵巢癌在体内的生长,进一步说明了CD59在肿瘤免疫逃逸中的作用。
OBJECTIVE: To study the silencing effect of CD59-siRNA on CD59-xenograft in ovarian cancer and its anti-tumor effect by in vivo animal experiment and to explore the role of CD59 in tumor immune escape. Methods: A2780 cells were transfected with CD59 interfering plasmid (group T) and empty plasmid (group V) by lipofectamine. The stable cell line was obtained. A2780 cells in the first two groups and untransfected A2780 cells were inoculated subcutaneously in nude mice to establish a tumor model. Tumor growth curves were drawn. The anti-tumor effect and silencing effects on CD59 were studied by RT-PCR and Western Blot. Results: The tumor growth curve showed that compared with the control group, the tumor growth of CD59 interference plasmid transfected group was significantly inhibited (P <0.05). The results of RT-PCR and Western Blot showed that CD59 mRNA and CD59 protein in the interference group were significantly lower than those in the control group (P <0.05). Conclusion: Animal experiments showed that siRNA targeting CD59 silencing could significantly inhibit the expression of CD59 and the growth of ovarian cancer in vivo, and further elucidated the role of CD59 in tumor immune escape.