Mechanism Of Transcription Factor Tex10 Combined With STAT3 To Regulate Hepatocellular Carcinomametastasis

Objective:To investigate the clinical significance of Tex 10 expression in HCC patients and its effect on the invasion and migration of HCC cells and the progression of EMT,and to explore the specific regulatory mechanism of Tex 10 involved in HCC recurrence and metastasis.Method:(1)TCGA databaseand KM-Plotter database were used to analyze the expression level of Tex 10 in hepatocellular carcinomatissues and the correlation between Tex 10 and hepatocellular carcinomaprognosis.(2)CCK-8 assay,soft AGAR colony formation assay and animal subcutaneous tumor model experiment verified the effects of Tex 10 overexpression on the growth and proliferation,colony formation and in vivo subcutaneous tumorigenicity of Huh7 and HepG2 hepatoma cells.(3)The effects of overexpression of Tex 10 on the invasion,migration and in vivo lung metastases of Huh7 and HepG2 cells were detected by Transwell assay and in vivo lung metastasis model experiments in mice.(4)Protein changes of EMT-related protein molecules N-cadherin,E-cadherin and Snail after Tex 10 overexpression were detected by Western Blot(WB).(5)Tex 10 was overexpressed in hepatocellular carcinoma cell lines,the changes of downstream target proteins in STAT3 signaling pathway were detected by WB assay,the interaction between Tex 10 and STAT3 was verified by Immunoprecipitation(IP)assay,and the role of p300 in this process was explored by WB assay.Result:(1)Tex 10 is highly expressed in HCC tissues and cells,and the overall survival,disease free survival and progression free survival of HCC patients with higher Tex 10 expression are significantly shorter than those with lower Tex 10 expression;(2)CCK-8 assay,soft AGAR colony formation assay and animal subcutaneous tumor model experiment showed that the growth and proliferation of Tex 10 overexpressed HCC cells increased,the number of colony formation increased,the volume and weight of subcutaneous tumor increased.(3)Transwell experiment and animal lung metastasis model experiment showed that the overexpression of Tex 10 promoted the invasion,migration and lung metastasis of hepatocellular carcinoma cells Huh7 and HepG2.(4)WB results showed that N-cadherin and Snail expression were up-regulated and E-cadherin expression was down-regulated after Tex 10 overexpression.(5)Overexpression of Tex 10 promoted the expression levels of CyclinD1,C-Myc and Bcl-XL downstream target proteins of STAT3 signaling path way;(6)Immunoprecipitation results showed that Tex 10 interacts with STAT3.(7)Overexpression of Tex 10 promotes the activeacetylation of STAT3;(8)When Tex 10 was overexpressed and p300 was silenced by siRNA,the expressions of CyclinD1,C-Myc and Bcl-XL downstream target proteins of STAT3 signaling pathway were down-regulated.Conclusion:(1)Tex 10 is highly expressed in HCC tissues and is positively correlated with poor prognosis of HCC patients,suggesting that Tex 10 can be an independent prognostic biomarker for HCC patients and may play an important role in the occurrence,development,recurrence and metastasis of HCC.(2)Overexpression of Tex 10 in vitro and in vivo can promote the growth and proliferation of HCC cells.(3)Overexpression of Tex 10 in vitro promotes the invasion and migration of HCC cells;In vivo overexpression of Tex 10 promotes lung metastasis of HCC cells.(4)Overexpression of Tex 10 promotes EMT process in HCC cells.(5)Overexpression of Tex 10 activates the expression of STAT3 signaling pathway and downstream target proteins.(6)Immunoprecipitation results showed that Tex 10 interacts with STAT3.(7)Tex 10 promotes the active acetylation of p300 substrate STAT3.(8)After silencing p300,Tex10’s up-regulation of downstream target proteins in STAT3 signaling pathway disappeared,indicating that Tex10’s regulation of STAT3 signaling pathway is mediated by p300.