【摘要】 目的探讨siRNA沉默细胞周期相关激酶(CCRK)基因对鼠结肠癌生长及其凋亡的影响。方法将50只BALB/C(nu/nu)裸鼠皮下接种人类结肠癌SW480细胞,将其中建立移植瘤裸鼠模型成功的30只裸小鼠按照单纯随机抽样方法随机分为3组,分别接种siRNA CCRK转染的人类结肠癌SW480细胞株(实验组)、正常培养的SW480细胞(空白对照组)和以转染携带无关序列片段sh RNA慢病毒的SW480细胞(阴性对照组)。第42天处死裸鼠后取下肿瘤组织,比较各组肿瘤重量、CCRKmRNA表达量、CCRK蛋白表达和细胞凋亡的变化。结果转染siRNA组肿瘤重量、CCRK mRNA和CCRK蛋白表达[(0.54±0.15)g,1.14±0.24和0.18±0.05]表达显著低于空白对照组[(1.05±0.14)g,2.41±0.42和0.49±0.07]和空siRNA载体组[(1.07±0.12)g,2.39±0.47,0.47±0.09;F=21.18,23.47,90.03;P<0.01];空白对照组与空siRNA载体组比较差异无统计学意义(q=0.98,1.07,1.13;P>0.05)。实验组沉默CCRK基因后组织中细胞凋亡率为(21.23±1.12)%,明显高于空白对照组(6.78±0.37)%和阴性对照组(7.25±0.32)%,差异有统计学意义(F=56.936,P<0.01);但空白对照组和空siRNA载体组之间差异无统计学意义(q=1.78,P>0.05)。结论 CCRK靶向siRNA表达载体接种结肠癌裸鼠后能显著降低结肠癌瘤体的增长,抑制CCRK基因和蛋白表达;siRNA沉默CCRK基因能促进其凋亡,CCRK基因有望成为结直肠癌治疗的新靶点。更多还原

【Abstract】 Objective To investigate the effect of siRNA silencing cell cycle-related kinase(CCRK) gene on the growth and apoptosis of colon cancer in mice,and to provide a new idea and method for the treatment of colon cancer.Methods Fifty BALB/C(nu/nu) nude mice were inoculated with human colorectal carcinoma SW480 cells,in which 30 nude mice were successfully established for transplantation tumor model and were randomly divided into three groups,siRNA CCRK transfection of human SW480 colon cancer cell line(experimental group) and normal cultured SW480 cells(blank control group),and carrying irrelevant sequence fragment shRNA slow transfection of virus SW480 cells(negative control group)respectively.The nude mice were sacrificed and the tumor tissues were removed after 42 days.The tumor weight,CCRK mRNA expression,CCRK protein expression and apoptosis rate were compared between the groups.Results siRNA transfection group,tumor weight,CCRK mRNA and CCRK protein expression[(0.54±0.15) g,1.14±0.24,0.18±0.05] were significantly lower than those of the control group [(1.05±0.14) g,2.41±0.42,0.49±0.07] and empty vector of siRNA group [(1.07±0.12) g,2.39±0.47,0.47±0.09,F=21.18,23.47,90.03; P<0.01].There were no significant differences between the blank control group and the empty siRNA vector group(q=0.98,1.07,1.13; P>0.05).Cells apoptosis rate in siRNA transfection group(21.23±1.12) % was significantly higher than that in normal control group(6.78±0.37) % and transfected with empty vector of siRNA group(7.25±0.32) %(P<0.01); but there was no significant difference between the blank control group and the empty siRNA vector group(q=1.78,P>0.05).Conclusions CCRK targeting siRNA expression vector inoculation with colon cancer can significantly reduce the growth of colon cancer and inhibit the expression of CCRK gene and protein in nude mice; CCRK gene silencing by siRNA can promote the colon cancer cells apoptosis; thus,CCRK gene can serve as a new potential molecular target for rectal cancer in gene therapy by siRNA.更多还原