Expression Of Girdin In Primary Hepatocellular Carcinoma And Its Effect On Cell Proliferation And Invasion

Objective:To investigate the expression and the clinical significance of Girdin in primary hepatocellular carcinoma, and to examine the effects of RNA interference (RNAi) Girdin on the bioactivity of HCC HepG2and Huh7.5.1cell lines.Methods:(1) Immunohistochemistry method was used to detect the expression of Girdin in40HCC tissues and30matched adjacent tissue. The relationship between the expression of Girdin and pathological factors in HCC were analyzed using the statistical methods.(2) We designed and synthesized the specific small interfering RNA of Girdin, and transfected it into HepG2and Huh7.5.1cells respectively by LipofectamineTM2000. Cells were transfected with no significant homology human gene sequences (negative control group) and without transfection treatment (blank control group) were treated in parallel. Real time-PCR was performed to determine the effects of siRNA on Girdin expression. Proliferation was assessed by MTT assay. Invasion abilities were determined by Transwell assay in vitro.Result:(1) Positive expression rate of Girdin in HCC tissues was higher than that in liver non-tumor tissue (P<0.001). The expression of Girdin protein was correlated with tumor size, T stage, TNM stage and Edmondson-Steiner stage (all P<0.05). The one-year and three-year cumulative survival rate for patients with low Girdin expression were100%and76.9%, which are significantly better than patients with high Girdin expression in these two groups (59.3%and14.8%)(P=0.001) Univariate analysis showed that the OS of HCC patient was correlated with T stage, TNM stage, Edmondson-Steiner stage and the expression of Girdin (P<0.05). Multivariate analysis displayed that tumor location, TNM stage, Edmondson-Steiner stage or the expression of Girdin could be the independent risk factors for the prognosis of HCC patients.(2) Real-time PCR showed that the specific Girdin siRNA significantly inhibited Girdin expression. And MTT assay revealed that the Girdin siRNA group significantly reduced cell proliferation comparing to the negative group or blank group cells (P<0.01). Transwell assay exhibited that HepG2and Huh7.5.1cells transfected with Girdin siRNA had much lower invasive activity than the negative group or blank group cells (P<0.05).Conclusion:(1) Girdin is highly expressed in HCC tissues. It is closely correlated with tumor size, T stage, TNM stage and Edmondson-Steiner stage. The expression of Girdin can be used as an independent adverse prognosticator of HCC.(2) Downregulation of Girdin by siRNA could effectively inhibit the growth of HepG2and Huh7.5.1cells. It also inhibits their invasion abilities.