着丝粒蛋白F对结直肠癌转移能力的影响  

Effect of centromere protein F on metastasis of colorectal cancer

作　　者：润增慈 魏辰[2] 王振雷 李剑[1] Run Zengci;Wei Chen;Wang Zhenlei;Li Jian(Department of General Surgery,He’nan Cancer Hospital,Affiliated Tumor Hospital of Zhengzhou University,Zhengzhou 450003,China;Department of Gastroenterology,He’nan Cancer Hospital,Affiliated Tumor Hospital of Zhengzhou University,Zhengzhou 450003,China)

机构地区：[1]河南省肿瘤医院普外科,郑州450003 [2]河南省肿瘤医院消化内科,郑州450003

出　　处：《中华实验外科杂志》2024年第8期1743-1746,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨过表达着丝粒蛋白F(CENPF)对结直肠癌侵袭和转移等恶性生物学行为的影响。方法购自中国科学院上海细胞库的结肠癌细胞系SW480,HCT116、LOVO和正常人肠上皮细胞HIEC-6培养至对数生长期,采用荧光定量聚合酶链反应(PCR)和蛋白质免疫印迹分析细胞CENPF mRNA和蛋白质表达水平。选取高表达CENPF细胞系HCT116作为研究对象,采用短发卡RNA(shRNA)对照慢病毒和CENPF shRNA慢病毒、空载慢病毒和CENPF过表达慢病毒感染HCT116细胞系,分别为shRNA对照组和CENPF shRNA组,采用划痕实验和Transwell实验分析两组细胞迁移和增殖能力;采用蛋白质免疫印迹分析两组细胞上皮-间充质转化能力。组间计量数据比较采用独立样本t检验。结果人肠上皮细胞HIEC-6 CENPF mRNA和蛋白质表达水平(1.04±0.09、0.50±0.10)明显低于结肠癌细胞系SW480,HCT116、LOVO(1.53±0.13、1.82±0.17、1.61±0.11;0.75±0.09、1.07±0.13、0.82±0.09),差异有统计学意义(t=7.619、10.230、10.040,P<0.05;t=4.565、8.631、5.817,P<0.05)。shRNA对照组细胞24、36、48 h吸光值明显高于CENPF shRNA组,差异有统计学意义(F=5.124,P<0.05)。对照组细胞24、36、48 h吸光值明显低于CENPF组细胞,差异有统计学意义(F=3.489,P<0.05)。shRNA对照组细胞24 h划痕愈合率和侵袭数量[(54.68±8.03)%、(114.75±7.71)个]明显高于CENPF shRNA组[(26.90±3.41)%、(71.00±8.44)个],差异有统计学意义(t=7.794、9.378,P<0.05)。对照组细胞24 h划痕愈合率和侵袭数量[(41.82±5.87)%、(105.33±21.59)个]明显低于CENPF组细胞[(70.62±6.31)%、(143.67±13.90)个],差异有统计学意义(t=8.185、3.387,P<0.05)。shRNA对照组细胞E-钙黏蛋白表达水平(1.02±0.08)明显低于CENPF shRNA组(1.35±0.06),差异有统计学意义(t=8.172,P<0.05)。shRNA对照组细胞N-钙黏蛋白和波形蛋白表达水平(0.77±0.09、0.83±0.07)明显高于CENPF shRNA组(0.44±0.06、0.42±0.09),差异有统计学意义(t=7.154,8.667,P<0.05)ObjectiveTo investigate the effects of overexpression of centromere protein F(CENPF)on invasion and metastasis of colorectal cancer and its molecular mechanism.MethodsColon cancer cell lines SW480,HCT116,LOVO and normal intestinal epithelial cells HIEC-6,purchased from Shanghai Cell Bank of Chinese Academy of Sciences,were cultured to logarithmic growth stage,and the mRNA and protein expression levels of CENPF were analyzed by fluorescent quantitative polymerase chain reaction(PCR)and Western blotting.The HCT116 cell line with high CENPF expression was selected as the study object,and the HCT116 cell line was infected with short hairpin RNA(shRNA)control lentivirus and CENPF shRNA lentivirus,empty lentivirus and CENPF overexpression lentivirus,respectively,as shRNA control group and CENPF shRNA SHRNA group.The ability of cell migration and proliferation was analyzed by scratch test and Transwell test.The epithelial mesenchymal transformation ability of the two groups of cells was analyzed by Western blotting.T test was used to compare measurement data between groups.ResultsThe mRNA and protein expression levels of HIEC-6 CENPF in human intestinal epithelial cells(1.04±0.09,0.50±0.10)were significantly lower than those of colon cancer cell lines SW480,HCT116 and LOVO(1.53±0.13,1.82±0.17,1.61±0.11;0.75±0.09,1.07±0.13,0.82±0.09,t=7.619,10.230,10.040,P<0.05;t=4.565,8.631,5.817,P<0.05).The cell absorbance values at 24,36 and 48 h in shRNA control group were significantly higher than those in CENPF shRNA group(F=5.124,P<0.05).The cell absorbance values at 24,36 and 48 h in the control group were significantly higher than those in CENPF group(F=3.489,P<0.05).The 24-h scratch healing rate and invasion number of cells in shRNA control group[(54.68±8.03)%,(114.75±7.71)cells]were significantly greater than those in CENPF shRNA group[(26.90±3.41)%,(71.00±8.44)cells,t=7.794,9.378,P<0.05].The 24-h scratch healing rate and invasion number of cells in control group[(41.82±5.87)%,(105.33±21.59)cells]were significant

关 键 词：着丝粒蛋白F 结直肠癌 迁移 侵袭 上皮-间充质转化 

分 类 号：R735.34[医药卫生—肿瘤]