Effect Of Overexpression Of CYFIP1 On Proliferation And Migration Of Nasopharyngeal Carcinoma Cell

Objective Tumor suppressor gene is closely related to the development of tumor.Abnormal expression of tumor suppressor gene can affect the biological function of tumor cells.In the past study,we found that the candidate tumor suppressor gene CYFIP1 in nasopharyngeal carcinoma and nasopharyngeal carcinoma cell lines were in the low expression or in lack of expression.This research was aimed to investigate the effect of overexpression of CYFIP1 on proliferation and migration of nasopharyngeal carcinoma cell.Methods 1.The lentiviral vectors containing CYFIP1 gene fragment and empty fragment were designed and packaged.A human nasopharyngeal carcinoma cell line(CNE2)was infected with a lentiviral vector containing CYFIP1,or the empty vector which were used as experimental group and blank group.The uninfected CNE2 acted as a control group.Cell stability was obtained by puromycin screening.2.The expression levels of CYFIP1 gene andprotein in different groups were measured by Western Blotting and qRT-PCR(quantitative real-time polymerase chain reaction).3.Cell migration was analyzed by transwell chamber and wound healing assays.The cell migration distance and the number of penetrating cells were calculated.The expression of E-cadherin gene was detected in different groups of cells.4.Cell proliferation was evaluated by MTT(represented by OD value)after cell culture for 24,48,72 and 96 h,separately.In clone formation assays,the cell culture were terminated when clones in plate could be seen by naked eyes and clones were counted.PI reagent was used to stain the cells,and the cell cycle was detected by flow cytometry.Results 1.The expression levels of mRNA and protein of CYFIP1 in experimental group,blank group and control group group were 3.491 ± 0.169,1.079 ± 0.037,1.00 ± 0.01 and 0.326±0.018?0.169±0.013?0.149±0.011,respectively.Compared with the control group and blank group,the expression of gene and protein level of CYFIP1 in the experimental group were significantly higher(P <0.05).The difference was statistically significant.2.In the observation of the effect of CYFIP1 on the migration ability of nasopharyngeal carcinoma cells,the cell migration distance of experimental group,blank group and control group were(6.71±0.1),(9.82±0.12)and(9.98±0.1)mm.Compared with blank group and control group,the experimental group had a lower migration distance(P < 0.05).The numbers of penetrating cells of experimental group,blank group and control group were(37.33±1.53),(62.67±5.03)and(62.13±4.36).The experimental group had a lower number of penetrating cells than blank group and control group.3.Comparing the OD value of different cell-culture times,three groups detected no significant changes(P > 0.05).The expression level of mRNA of E-cadherin inexperimental group,blank group and control group group were 1.316±0.166?1.070±0.033 ? 1.00±0.01,respectively.Compared with the control group and blank group,the expression of E-cadherin in the experimental group were significantly higher(P <0.05).The number of clone of experimental group,blank group and control group were(414±29),(426±30)and(381±59).The difference was not statistically significant(P > 0.05).In the cell cycle detection,the S-phase cells of experimental group,blank group and control group were(23.773 ± 3.586)%,(22.253 ± 2.089)% and(24.963 ± 1.896)%,and there was no significant difference between the different cell groups(P> 0.05).Conclusions 1.The CYFIP1-CNE2 cell with stable expression is successfully constructed.2.There was no significant correlation between CYFIP1 gene expression and tumor proliferation in nasopharyngeal carcinoma.However,CYFIP1 is related to tumor cell migration.CYFIP1 maybe participate in the infiltration and development of nasopharyngeal carcinoma,which is expected to become a molecular marker of nasopharyngeal carcinoma metastasis.