| Part OneObjective: to investigate the effect of KIF2C gene on the prognosis of patients with primary hepatocellular carcinoma.Methods:first,we obtained complete survival data and Gene Expression Omnibus(GEO)data from the high-throughput Expression database according to the inclusion criteria(1)for primary hepatocellular carcinoma(HCC),(2)and(3).The filtered GSE14520 dataset was updated to April 5,2017 and validated on December 15,2017.The data were downloaded and the gene expression data set of primary hepatocellular carcinoma was obtained.The gene expression level of KIF2C was screened from the data for data processing.Univariate analysis was performed on the general clinical data,and the indicators with statistical differences(P< 0.05)were included in the multivariate analysis.Meanwhile,kaplan-meier method was used to draw the survival analysis curve to explore the relationship between the expression level of KIF2C and the prognosis of patients with hepatocellular carcinoma associated with hepatitis b.Then we by The Cancer Genome Atlas(TCGA)database search of primary hepatocellular carcinoma(HCC)in patients with clinical information and gene expressionprofile,download The data and according to The same into The group of 370 cases of patients with primary liver Cancer screening with standard data and The gene expression patterns,carries on The single factor index and statistically significant(P<0.05)in multi-factor analysis.Kaplan-meier method was also used to draw the survival analysis curve.Results: the multivariate analysis and survival curve of the two groups were statistically different(P < 0.05).Results: multivariate analysis of GEO data showed that the expression of KIF2C was correlated with the prognosis of patients(P=0.009,HR(95%ci)=1.816(1.159-2.844)),and kaplan-meier survival analysis showed poor prognosis of patients with high KIF2C expression(P=0.0119).Multivariate analysis of TCGA data showed that the expression level of KIF2C was correlated with the prognosis of patients(P=0.001,HR(95%ci)=1.949(1.324-2.869)),and kaplan-meier survival analysis showed poor prognosis of patients with high KIF2C expression(P< 0.0001).Conclusion: the level of KIF2C expression has a certain impact on the prognosis of patients with primary hepatocellular carcinoma,and patients with high expression have poor prognosis.It may provide new ideas for the targeted treatment of primary hepatocellular carcinoma.Part TwoObjective: to investigate the effect of KIF2C gene on the biological behavior of SMMC-7721 hepatocellular carcinoma cell line.Methods: the expression of KIF2C in various liver cancer cell lines was detected by real-time fluorescence quantitative PCR,and appropriate cell lines were selected for subsequent study.Then the smmc-7721 cell line was transfected with lentivirus mediated technology to construct the transfected cell line with KIF2C gene silencing and overexpression stability,and the silencingand overexpression efficiency of KIF2C gene in ssmc-7721 cell line was detected by western blot.In vitro functional tests of KIF2C stable transfected cell lines were conducted,including cell proliferation experiment,migration experiment,cell plate clone formation experiment,invasion experiment,etc.,to observe the effect of KIF2C gene on the biological behavior of smmc-7721 liver cancer cell line.Results: in the cell proliferation experiment,the number of smmc-7721 and smmc-7721-kif2c-overexpressed cells was significantly higher than that of smmc-7721-kif2c-silenced cells(P=0.011,P=0.035)in the cells reseeded and cultured for 24 h(P=0.004),48 h(P< 0.001)and 72 h(P < 0.001).The clonal formation rate of smmc-7721-kif2c-overexpressed cells was significantly higher than that of smmc-7721,smmc-7721-nc and smmc-7721-kif2c-silenced cells.In the cell scratch experiment,the average migration distance of smmc-7721-kif2c-overexpressed cells within 0-24 h and 24-48 h was significantly higher than that of smmc-7721-kif2c-silenced cells(P=0.044,P=0.016).In thecell invasion experiment,the number of smmc-7721-kif2 coverexpressed cells crossing the stromal membrane was significantly higher than that of smmc-7721,smmc-7721-nc and smmc-7721-kif2c-silenced cells(P=0.022,P=0.002,P< 0.001).Conclusion: KIF2C gene can promote the proliferation,migration and invasion of smmc-7721. |
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