EGFR shRNA联合雷帕霉素对Colo-16细胞增殖、迁移及侵袭的影响  

Effects of EGFR shRNA combined with rapamycin on proliferation, migration and invasion of Colo-16 cells

作　　者：王慧 杨艳 李竹茜 展晓红[2] 魏志平 WANG Hui;YANG Yan;LI Zhuqian;ZHAN Xiaohong;WEI Zhiping(Department of Dermatology,Chengyang District People s Hospital,Qingdao 266109,China;Department of Pathology,Affiliated Hospital of Qingdao University,Qingdao 266109,China;Department of Dermatology,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,China)

机构地区：[1]青岛市城阳区人民医院皮肤科,青岛266109 [2]青岛大学附属医院病理科,青岛266109 [3]徐州医科大学附属医院皮肤科,徐州221002

出　　处：《中国麻风皮肤病杂志》2023年第7期479-484,共6页China Journal of Leprosy and Skin Diseases

摘　　要：目的:明确表皮生长因子受体(EGFR)shRNA联合雷帕霉素(RAPA)对皮肤鳞癌Colo-16细胞增殖、迁移和侵袭的影响,并对其可能机制进行初步探究。方法:Colo-16细胞分为5组:正常细胞组(Colo-16细胞+磷酸盐缓冲液PBS)、阴性对照组(转染shRNA-NC质粒的Colo-16细胞+PBS)、EGFR shRNA组(转染EGFR shRNA质粒的Colo-16细胞+PBS)、RAPA组(Colo-16细胞+RAPA)、联合组(转染EGFR shRNA质粒的Colo-16细胞+RAPA)。采用细胞计数试剂盒(CCK-8)和克隆形成实验测定细胞增殖能力;划痕实验和Transwell法检测细胞迁移和侵袭情况,采用Western blot、RT-PCR检测增殖、侵袭相关基因Ki-67、MMP-2、MMP-9蛋白和mRNA水平表达。多组间比较采用单因素方差分析,组间两两多重比较采用SNK-q检验。结果:EGFR shRNA组、RAPA组及联合组Colo-16细胞吸光度A值、细胞克隆形成率、划痕愈合率、侵袭细胞数目、Ki-67、MMP-2、MM-P9等基因蛋白和mRNA表达显著低于正常细胞组与阴性对照组,差异有统计学意义(均P<0.05),且联合的效果更加明显。正常细胞组与阴性对照组差异均无统计学意义(均P>0.05)。结论:EGFR shRNA联合雷帕霉素在抑制Colo-16细胞增殖、迁移及侵袭方面有协同增效的作用,其机制可能与EGFR/PI3K/AKT/mTOR信号通路的双重抑制作用相关。Objective:To investigate the effect of epidermal growth factor receptor(EGFR)shRNA combined with rapamycin(RAPA)on the proliferation,migration and invasion of skin squamous cell carcinoma Colo-16 cells,and to explore its possible mechanism.Methods:Colo-16 cells were divided into five groups,normal cell group(Colo-16+phosphate buffer saline PBS),negative control group(Colo-16 cells+PBS transfected with shRNA-NC plasmid),EGFR shRNA group(Color-16 cells+PBS transfected with EGFR shRNA plasmid),RAPA group(conventionally cultured Colo-16 cells+RAPA),and combination group(Colo-16 cells+RAPA transfected with EGFR shRNA plasmid).Cell proliferation ability was measured by cell counting kit(CCK-8)and clone formation test.Scratch test and Transwell method were used to detect cell migration and invasion.The expression of Ki-67,MMP-2,MMP-9 protein and mRNA related to proliferation and invasion were detected by Western blot and RT-PCR.Single factor analysis of variance was used for comparison among multiple groups,and SNK-q test was used for multiple comparison between groups.Results:Colo-16 cell absorbance A value,cell clone formation rate,scratch healing rate,number of invasive cells,Ki-67,MMP-2,MM-P9 gene protein and mRNA expression in EGFR shRNA group,RAPA group and combination group were significantly lower than those in normal cell group and negative control group,with significant differences(P s<0.05),and the combination effect was more obvious.There was no statistically significant difference between normal cell group and negative control group(P s>0.05).Conclusion:EGFR shRNA combined with rapamycin has synergistic effect in inhibiting the proliferation,migration and invasion of Colo-16 cells,and its mechanism may be related to the double inhibition of EGFR/PI3K/AKT/mTOR signal pathway.

关 键 词：皮肤鳞状细胞癌 表皮生长因子 RNA干扰 雷帕霉素 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R739.5[医药卫生—肿瘤]