UbcH10基因沉默对裸鼠移植性大肠癌抑制作用

目的 通过短发夹RNA(shRNA)干扰沉默UbcH10基因的表达,并观察其对裸鼠移植结直肠癌的抑制作用。方法 构建RNA干扰表达载体pGPU6/GFP/Neo/UbcH10-RNAi(pUbcH10-RNAi),建立UbcH10基因沉默细胞系HT-29/UbcH10-RNAi;采用蛋白印迹法(western blot)评价pUbcH10-RNAi对UbcH10基因表达的影响,观察移植结直肠癌裸鼠的肿瘤生长情况。结果 HT-29/UbcH10-RNAi细胞中UbcH10基因表达明显下调;与对照组比较HT-29/UbcH10-RNAi组肿瘤生长明显减慢,在接种肿瘤细胞12 d后隐约可见瘤体,36 d后断颈处死剥离瘤体重量仅是对照组肿瘤的40%左右。结论 短发夹RNA(shRNA)沉默UbcH10基因对裸鼠移植性大肠癌有抑制作用,提示抑制UbcH10基因表达可能是大肠癌治疗的潜在方法。

Inhibitory effect of UbcH10 gene silencing on colorectal cancer cell xenografts

Objective To silence ubiquitin conjugating enzyme-10(UbcH10) gene by RNA interference(RNAi) and to observe its inhibitory effect on colorectal cancer cell xenograft in nude mouse.Methods The expression vector pGPU6/GFP/Neo/UbcH10-RNAi(pUbcH10-RNAi) was constructed and the UbcH10 gene silencing cell lines HT-29/UbcH10-RNAi was established.Western blot analysis was used to evaluate the expression of the UbcH10 gene.The inhibitory effect of pUbcH10-RNAi was evaluated in a nude mouse colorectal cancer cell xenograft model.ResultsThe results demonstrated that UbcH10 gene expression was significantly decreased in pUbcH10-RNAi treated cells.HT-29/UbcH10-RNAi cells xenograft demonstrated growth suppression compared to that of the control.The size and weight of tumors in HT-29/UbcH10-RNAi groups were significantly decreased.On the 36th day after the inoculation,the average weight of tumors in HT-29/UbcH10-RNAi group was only about 40% of that in HT-29/RNAi-NC group and HT-29/PBS group(P<0.05).Conclusion Silencing of UbcH10 gene by shRNA has antitumor activity on nude mouse colorectal cancer cell xenograft model and may be a therapeutic potential for the treatment of colorectal cancer.