PKC信号通路在KISS1基因抑制人结直肠癌HCT116细胞侵袭迁移的作用  

Expression of KISS1 gene inhibits the human colorectal carcinoma cell HCT116 migration by PKC signaling pathway

作　　者：陈志华[1] 韩宏景 林素勇[1] 苏小宝[1] 戴起宝[1] 陈绍勤[1] 吴佩文[2] 

机构地区：[1]福建医科大学附属第一医院胃肠外科2区,福州350005 [2]福建医科大学附属第一医院内分泌科,福州350005

出　　处：《重庆医科大学学报》2017年第11期1448-1452,共5页Journal of Chongqing Medical University

基　　金：国家重点临床专科建设基金资助项目(编号:2013-GJLCZD);福建医科大学苗圃基金资助项目(编号:2014MP020);福建省教育厅B类科技项目(JB12108)

摘　　要：目的:探讨PKC信号通路在KISS1基因发挥抑制结直肠癌细胞HCT116侵袭迁移的作用。方法:构建p GC-LV-KISS1-EGFP慢病毒载体感染人结直肠癌细胞HCT116,分空白对照组(CON组)、空载体对照组(NC组)、过表达组(OE组)。q RT-PCR和Western blot检测KISS1基因m RNA和Metastin、PKC信号通路的关键分子PKCα、PKCβII及下游E钙黏蛋白表达量的变化。Transwell法检测细胞侵袭、迁移能力的变化。结果:转染后能够稳定表达报告基因EGFP,荧光率均>80%,且OE组KISS1基因m RNA、Metastin表达量较CON组和NC组均明显升高(P<0.05)。转染后,PKCβⅡ的蛋白表达量(0.288 2±0.023 7)较CON组(0.530 9±0.013 3)和NC组(0.511 7±0.008 5)明显下降(P<0.05),而PKCα的表达水平无明显变化(P>0.05);下游效益蛋白E钙黏蛋白表达量(0.633 2±0.017 4)较CON组(0.232 2±0.019 6)和NC组(0.252 3±0.018 2)明显升高(P<0.05)。OE组细胞侵袭、迁移能力较CON组和NC组均明显下降(P<0.05)。结论:KISS1基因可能通过下调PKCβⅡ后上调下游蛋白E钙黏蛋白的表达而发挥抑制结直肠癌细胞HCT116侵袭、迁移作用,有望成为防治结直肠癌转移的新靶点。Objective：To investigate the effect of KISS1 gene suppression on the metastatic capacity of the human colorectal carcinoma cells HCT116 and the involvement of protein kinase C signaling pathway. Methods：pGC-LV-KISS1-EGFP lentiviral vector was constructed and the colorectal cancer cells HCT116 was infected and divided into：the point blank control group（CON）,the non- cartier contral group（NC） and the over-expression group（OE）, qRT-PCR and Western blot methods were used to detect KISS1 gene product mRNA and Metastin level, the key molecular of PKC signaling pathway PKCα,PKCβ 11 and E-cadherin sticky protein expression. Transwell Chambers methods were used to detect the changes of cell＇s invasion and migration ability. Results：After lentiviral transfection, HCT-116 ceils could stably express EGFP report gene,and the fluorescence rate was over 80%. Compare with those of the CON and NC, the KISS1 gene product mRNA and Metastin level of the OE were significantly increased. The protein expression of PKCβ II （0.288 2 ＋ 0.023 7） was significantly increased compared with that of the CON（0.530 9 ± 0.013 3） and NC （0.511 7 ± 0.008 5,P〈0.05）. But the protein expression of PKCα had no significant change（P〉0.05）. The E-cadherin expression level（0.633 2 ± 0.017 4） rose highly compared with that of the CON（0.232 2 ± 0.019 6） and NC（0.252 3 ± 0.018 2） with statistical significances（P〈0.05）. The cell＇s invasion,migration ability of the OE group also significantly decreased compared with that of the other two groups（P〈0.05）. Conclusion ：KISS1 gene may regulate the PKCβⅡ and raise the E-cadherin sticky protein expression and plays a role of inhibiting the invasion and migration in the HCT116,which will become a new target for the prevention and treatment of cancer metastasis.

关 键 词：结直肠癌 KISS1基因 PKC 信号通路 转移 

分 类 号：R735.34[医药卫生—肿瘤]