The Clinical Significance Of KIF2C Gene Expression Level In Hepatocellular Carcinoma And Effect On Cell Proliferation,Cell Cycle,Apoptosis

Objective: This study aimed at exploring the clinical significance of KIF2 C expression level in hepatocellular carcinoma and the effect on cell proliferation,cell cycle,apoptosis.Methods: The differential expression of KIF2 C in hepatocellular carcinoma and the correlation of gene expression with clinicopathological variables were investigated based on the public databases by bioinformatics analysis methods.The hepatocellular carcinoma cell lines Hep3 B and Huh7 were transfected by the virus vector to up-regulate or down-regulate the expression level of KIF2 C gene.The up-regulation and down-regulation efficiency of KIF2 C at mRNA level in the stable transfected cell lines were validated by q RT-PCR method.CCK8 assay was performed to detect the proliferation ability of Hep3 B and Huh7 in vitro.Flow cytometric analysis was used to detect the effect on cell cycle and cell apoptosis.Transcriptome sequencing was used to screen for significantly differentially expressed miRNAs and enrichment analysis of their target genes was conducted to screen related biological processes and molecular signaling pathways.Results: 1.The expression level of KIF2 C in tumor tissue was obviously higher than adjacent non-tumor tissue(P < 0.05).In addition,the expression level of KIF2 C gene was significantly associated with serum AFP level,tumor stage,and pathological grade(P < 0.05).2.Comparing with the normal group,the mRNA level of KIF2 C gene was significantly up-regulated or down-regulated in over-expression group or silenced group,correspondingly(P < 0.05).The above results suggested that we had successfully transfected the cells and interfered the expression level of target gene.3.CCK8 assay was performed to detect the effect on cell proliferation ability after the KIF2 C expression level had been interfered: comparing with the normal group,the proliferation ability was enhanced substantially in the over-expression group,meanwhile,the proliferation ability was significantly inhibited in silenced group(P < 0.05).4.The results of cell cycle suggested that: the cell cycle process was not notably altered in the over-expression group(P > 0.05).In contrast,the cell cycle process was remarkably blocked and the proportion in G0/G1 phase was significantly increased after the expression of KIF2 C gene being interfered(P <0.05).5.The results of cell apoptosis showed that: the distribution proportion of apoptosis cell in the over-expression group was not significantly altered(P > 0.05).While,in the silenced group,the proportion of apoptosis cell was substantially increased,especially for the early-stage apoptosis cell(P < 0.05).6.Among all the up-regulated genes,the most obvious differentially expressed gene is miRNA-219a-2-3p;among all the down-regulated genes,the most significant differentially expressed gene is miRNA-4286.Enrichment analysis showed that the target genes of miRNA are mainly enriched in lipid metabolism pathway,Ras signaling pathway,MAPK signaling pathway,and T cell receptor signaling pathway.Conclusion: KIF2 C gene can promote the progression of hepatocellular carcinoma and is associated with its malignant characteristic.Down-regulated KIF2 C gene expression could significantly inhibit cell proliferation,delay cell cycle process,and promote cell apoptosis.