重组质粒pCMV AIM-1 antisense对卵巢癌裸鼠移植瘤的抑瘤效应研究

发布时间：2018-03-10 13:11

本文选题：重组质粒　切入点：AIM-　出处：《肿瘤防治研究》2015年11期 　论文类型：期刊论文

【摘要】：目的探讨pCMV AIM-1 antisence重组质粒对卵巢癌裸鼠移植瘤的体内抑瘤效应。方法(1)人卵巢癌细胞A2780转染pCMV空载体和pCMV AIM-1 antisense重组质粒,G418筛选出抗性克隆,扩增后产生的稳定转染株用于实验;(2)通过RT-PCR、Western blot检测未转染A2780细胞(对照组)、A2780转染pCMV空载体和pCMV AIM-1 antisense重组质粒三组AIM-1 m RNA和蛋白的表达;(3)将稳定转染的pCMV空载体和pCMV AIM-1 antisense重组质粒的A2780细胞注入裸鼠皮下,建立裸鼠移植瘤模型,未转染A2780细胞注入裸鼠皮下作为对照,观察三组裸鼠移植瘤生长情况,计算肿瘤重量和肿瘤体积,并进行统计对比分析;(4)SP免疫组织化学法检测移植瘤组织切片AIM-1的表达。结果 (1)A2780对照组和转染pCMV空载体组AIM-1 m RNA和蛋白表达无明显改变;而A2780转染pCMV AIM-1 antisense重组质粒组细胞AIM-1 m RNA和蛋白表达明显下降;(2)A2780对照组、A2780转染pCMV空载体组和A2780转染pCMV AIM-1 antisense重组质粒组的裸鼠肿瘤重量分别为(1.93±0.21)g、(1.90±0.39)g和(0.77±0.34)g;移植瘤体积分别为(1 011.7±235.6)mm3、(981.2±162.3)mm3和(468.7±119.6)mm3,重组质粒组与空载体组和对照组相比,差异有统计学意义(P0.01);pCMV AIM-1 antisense重组质粒的抑瘤率达53.8%。(3)SP免疫组织化学法检测瘤体组织切片AIM-1的表达情况,见A2780对照组和转染pCMV空载体组AIM-1表达没有明显改变,而A2780转染pCMV AIM-1antisense重组质粒组AIM-1的表达明显减少。重组质粒组AIM-1蛋白表达阳性率和阳性标记指数均明显低于对照组和空载体组(P0.01)。结论 pCMV AIM-1 antisense重组质粒对A2780移植瘤生长有明显抑制作用。
[Abstract]:Objective to investigate the inhibitory effect of pCMV AIM-1 antisence recombinant plasmid on ovarian cancer xenografts in nude mice. Methods: human ovarian cancer cell line A2780 was transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid G418 was used to screen the resistant clones. The stable transfection strain was used in the experiment. The untransfected A2780 cell line (the control group was transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid three groups AIM-1 m RNA and protein expression) was detected by RT-PCRG Western blot. A2780 cells of pCMV AIM-1 antisense recombinant plasmid were injected subcutaneously into nude mice. The transplanted tumor model of nude mice was established, and the untransfected A2780 cells were injected subcutaneously into nude mice as control. The growth of transplanted tumor was observed in three groups, and the tumor weight and volume were calculated. The expression of AIM-1 m RNA and protein in transplanted tumor sections were detected by immunohistochemical method. Results the expression of AIM-1 m RNA and protein in the control group and the empty pCMV vector group were not significantly changed. However, the expression of AIM-1 m RNA and protein in A2780 transfected pCMV AIM-1 antisense recombinant plasmid group was significantly decreased. The tumor weight of Nude mice transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid was 1.93 卤0.21 卤0.39 g and 0.77 卤0.34 g, respectively, and the tumor volume of transplanted tumor was 0.77 卤0.34 g, respectively, and the tumor weight of nude mice transfected with A2780 was 1.93 卤0.21 卤0.39 g and 0.77 卤0.34 g respectively, and that of A2780 transfected with pCMV AIM-1 antisense was 1.90 卤0.39 g and 0.77 卤0.34 g respectively. Compared with empty vector group and control group, 1011.7 卤235.6 mm ~ (-3) and 468.7 卤119.6 mm ~ (-3) mm3 and 468.7 卤119.6 mm ~ (-3) mm3 respectively, the recombinant plasmid group was compared with the empty vector group and the control group. The inhibitory rate of pCMV AIM-1 antisense recombinant plasmid was 53.8%. The expression of AIM-1 in tumor sections was detected by immunohistochemistry. There was no significant change in AIM-1 expression in A2780 control group and empty pCMV vector transfection group. The positive rate and labeling index of AIM-1 protein expression in the recombinant plasmid group were significantly lower than those in the control group and empty vector group (P 0.01). Conclusion the expression of pCMV AIM-1antisense recombinant plasmid in A2780 transplanted tumor was significantly lower than that in the control group and empty vector group. Growth has obvious inhibitory effect.
【作者单位】：武汉市中心医院中西医结合肿瘤科;武汉市中心医院影像诊断科;
【分类号】：R737.31

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