Expression Of GPC3 In Hepatocellular Carcinoma Cell Lines With Different Metastatic Invasion Ability And Its Relationship With 4E-BP1 And PS6K

Objective:Detection of phosphatidylinositol proteoglycan 3(GPC3),eukaryotic cell activating factor 4E binding protein 1(4E-BP1)and ribosomal S6 protein kinase(PS6K)in three hepatocellular carcinoma(HCC)cell lines with different metastatic capabilities m RNA and protein expression.Based on the analysis of bioinformatics,the correlation between these three genes and the metastasis and invasion ability of different hepatocellular carcinoma cells were analized.Methods:GPCR3 m RNA and protein expression levels of human normal liver cell line HL-7702,low metastatic hepatocellular carcinoma cell line MHCC-97 L,high metastatic hepatocellular carcinoma cell line MHCC-97 H were detected using fluorescent quantitative PCR and Western blotting methods,respectively.,and 4E-BP1 m RNA and protein expression levels,PS6 K m RNA and protein expression levels.The relationship between the three proteins and the invasion ability of hepatocellular carcinoma(HCC)was specifically analyzed by R language.Results: The online website cbioportal analysis showed that the prognosis of HCC patients with abnormal expression of GPC3,4E-BP1,and PS6 K was significantly different from that of HCC patients expressing this gene(p-values were 0.0281,1.18e-7,and 0.0119,respectively).The life cycle is reduced.Quantitative quantitative PCR results showed that compared with HL-7702 cell line,GPC3 and PS6 K m RNA levels expressed by MHCC-97 H cells were significantly higher than HL-7702(P<0.05),while GPC3 m RNA levels expressed by MHCC-97 L cells were significantly increased.High and statistically significant(P <0.05),the expression of PS6 K slightly increased but there was no significant difference.The m RNA expression of 4E-BP1 was HL-7702,MHCC-97 L,and MHCC-97 H from high to low,and there weresignificant differences compared with the HL-7702 group(* P <0.05).Compared with MHCC-97 L,MHCC-97 H significantly increased the levels of GPC3 and PS6 K m RNA,and significantly reduced the expression of 4E-BP1 m RNA(P <0.05).Western blot results also showed that compared with MHCC-97 L,the levels of GPC3 and PS6 k proteins expressed by MHCC-97 H cells were significantly increased and there were statistical differences(P <0.05).On the contrary,the expression level of 4E-BP1 protein was higher To the lowest are HL-7702,MHCC-97 L,and MHCC-97 H.According to the value of protein expression obtained by Western blot experiments,the correlation matrix analysis was performed by R programming with r> 0.7 as positive correlation and r <-0.7 as negative correlation.In three cell lines,PS6 k was positively correlated with GPC3 protein expression,with correlation coefficients greater than 0.8.4E-BP1 was negatively correlated with GPC3 protein expression,and correlation coefficients were less than-0.8.PS6 k was negatively correlated with4E-BP1 protein expression.The correlation coefficients are all less than-0.8.Conclusion:In three cell lines,the m RNA and protein levels of GPC3 and PS6 K increased with increasing metastatic capacity,and PS6 K was positively correlated with GPC3 expression.The expression of 4E-BP1 was inhibited with the increase of metastatic invasion ability,and was negatively correlated with the expression of GPC3.And the expression of these three proteins is closely related to the enhanced invasion ability of HCC.