摘要
目的研究慢病毒介导的连锁的泛素特异肽酶9(USP9X)基因RNA干扰对人胰腺癌细胞系裸鼠皮下移植瘤的影响。方法将靶向USP9X基因的干扰小RNA(siRNA)慢病毒(LV-siRNA-USP9X)和阴性对照慢病毒(LV-siRNA-NC)转染至SW1990细胞,并分别作为siUSP9X组和siNC组,未转染细胞作为对照组,分别将3组细胞系接种至裸鼠皮下,比较3组裸鼠接种第30天的移植瘤生长情况和瘤体重量。蛋白质印记(Western blot)法和免疫组织化染色检测3组裸鼠皮下移植瘤中survivin蛋白的表达情况,TUNEL法检测细胞凋亡指数(AI)。结果siUSP9X组USP9X蛋白的相对表达量均明显低于siNC组和对照组,差异均有统计学意义(P<0.01),干扰效率约为90%。siUSP9X组裸鼠的肿瘤体积明显小于siNC组和对照组裸鼠,且随着接种时间的延长,差异越来越明显。在接种第30天时,siUSP9X组裸鼠瘤体重量均小于siNC组和对照组裸鼠,抑瘤率为52%。siUSP9X组survivin蛋白的相对表达量和阳性表达率均低于siNC组和对照组,AI均高于siNC组和对照组,差异均有统计学意义(P<0.05)。结论慢病毒介导的USP9X基因沉默可明显抑制人胰腺癌细胞系裸鼠皮下移植瘤的生长,该现象可能与其下调survivin蛋白的表达并促进细胞凋亡有关,USP9X有可能成为胰腺癌基因治疗的一个新靶点。
Objective To investigate the effect of lentivirus-mediated RNA interference targeting ubiquitin specific peptidase 9 X-linked (USP9X) in the subcutaneous pancreatic cancer xenografts in nude mice.Method SW1990 cells were transfected with small interfering RNA (siRNA) lentivirus targeting USP9X (LV-siRNA-USP9X) and negative control lentivirus (LV-siRNA-NC) as siUSP9X group and siNC group, respectively, non-transfected cells were used as control, the nude mice were inoculated with the three stable cell lines to establish the subcutaneous tumor model of human pancreatic cancer, the growth and weight of xenografts on the 30th day were compared among the three groups. The expression of survivin protein was detected by Western blot and immunohistochemical staining, and apoptosis index (AI) was measured by TUNEL method. Result The relative expression of USP9X protein in siUSP9X group was significantly lower than that in siNC group and control group, the difference was statistically significant (P<0.01), the interference rate was about 90%. siUSP9X group had significantly lower tumor volume compared with siNC group and control group, furthermore, the difference was more obvious as the inoculation time elapsed. On the 30th day after inoculation, the tumor weight of siUSP9X group was markedly less than that in siNC group and control group, the tumor inhibition rate was 52%. The relative expression and positive rate of survivin protein in siUSP9X group was significantly decreased compared to siNC group and control group, while AI was higher instead, with statistically significant difference observed (P< 0.05). Conclusion Lentivirus-mediated RNA interference targeting USP9X can effectively inhibit the growth of the subcutaneous pancreatic xenografts in nude mice, which may be related to the down-regulation of survivin protein, and promotion of the cell apoptosis, USP9X gene may be an effective target for gene therapy in treating pancreatic cancer.
作者
周晓华
刘志毅
张蓬波
张秀忠
任泽强
孙文白
徐盼
吴耐
钱旭
ZHOU Xiaohua;LIU Zhiyi;ZHANG Pengbo;ZHANG Xiuzhong;REN Zeqiang;SUN Wenbai;XU Pan;WU Nai;QIAN Xu(Department of General Surgery, Nanjing General Surgery of Gaochun People’s Hospital, Nanjing 211300, Jiangsu, China;Graduate School, Xuzhou Medical University, Xuzhou 221004, Jiangsu, China;Department of General Surgery, Xuzhou Medical University, Xuzhou 221000, Jiangsu, China;Department of Pancreatic Surgery, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu, China)
出处
《癌症进展》
2019年第8期905-909,共5页
Oncology Progress
