Effect of centromere protein F on metastasis of colorectal cancer
Objective To investigate the effects of overexpression of centromere protein F(CENPF)on invasion and metastasis of colorectal cancer and its molecular mechanism.Methods Colon 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.Results The mRNA and protein expression levels of HIEC-6 CENPF in human intestinal epitheli-al 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 signifi-cantly higher than those in CENPF group(F=3.489,P＜0.05).The 24-h scratch healing rate and inva-sion number of cells in shRNA control group[(54.68±8.03)％,(114.75±7.71)cells]were signifi-cantly 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 significantly lower than those in CENPF group[(70.62±6.31)％,(143.67±13.90)cells,t=8.185,3.387,P＜0.05].The expression level of E-cadherin in shRNA control group(1.02±0.08)was significantly lower than that in CENPF shRNA group(1.35±0.06,t=8.172,P＜0.05).The expression levels of N-cadherin and vimentin in shRNA control group(0.77±0.09,0.83±0.07)were significantly higher than those in CENPF shRNA group(0.44±0.06,0.42±0.09,t=7.154,8.667,P＜0.05).The expression level of E-cadherin in control group(1.12±0.10)was significantly higher than that in CENPF group(0.63±0.07,F=10.140,P＜0.05).The ex-pression levels of N-cadherin and vimentin in control group(0.82±0.09,0.65±0.06)were significantly lower than those of CENPF shRNA group(1.39±0.09,1.18±0.11,t=11.230,10.350,P＜0.05).Conclusion The expression level of CENPF is significantly increased in colorectal cancer cells,and it is involved in cell biological behaviors related to metastasis such as migration and invasion of colorectal cancer cells.
Centromere protein FColorectal cancerMigrationTo invadeEpithelial mesenchymal transformation
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