摘要
Abstract
Objective To investigate the roles of the mechanosensitive ion channel protein Piezo1 and Yes-associated transcription pro-tein 1(YAP1)in the development of liver hepatocellular carcinoma(LIHC).Methods The expression levels of Piezo1 and YAP1 in LIHC,as well as their relationship with tumor grade,stage,lymph node metastasis,methylation level,and patient prognosis,were analyzed using the Tumor Immune Estimation Resource(TIMER)and UALCAN databases.The correlation between Piezo1 and YAP1 protein expression levels and their expressions in different types of liver cancer were analyzed using the Gene Expression Profiling Interactive Analysis(GEPIA)database.The gene interaction network between Piezo1 and YAP1 was analyzed using the GeneMANIA database.Small interfering RNA(si-RNA)was used to knock down Piezo1 in liver cancer cells MHCC97H and Hep3B.The expression levels of Piezo1,YAP1,nuclear factor κB(NF-κB)pathway-related proteins,and extracellular matrix(ECM)-related proteins were detected using quantitative real-time PCR and Western blot.Cell proliferation and invasion abilities after Piezo1 knockdown were evaluated using Cell Counting Kit-8(CCK-8)assay,transwell assay,and colony formation assay.Results The analysis of TIMER,UALCAN,and GEPIA databases showed that both Piezo1 and YAP1 were synergistically highly expressed in LIHC,and their expressions were associated with LIHC grade,stage,lymph node metastasis,and DNA promoter methylation(all P＜0.05),and correlated with poor prognosis in LIHC patients.GeneMANIA database analysis revealed that Piezo1 and YAP1 form an extensive gene interaction network,collectively promoting the infiltration of cancer-associated fibroblasts.PCR results indicated that Piezo1 and YAP1 were highly expressed in liver cancer cell lines MHCC97H,Hep3B,and HepG2(all P＜0.05).After knocking down Piezo1 in MHCC97H cells,the expression of YAP1 decreased,the phosphorylation of NF-κB inhibitor alpha(IκB-α)and p-NF-κB in the NF-κB signaling pathway was inhibited,and the expressions of ECM-related proteins fibronectin and collagen Ⅰ were reduced.Meanwhile,cell proliferation,invasion,and colony formation abilities were suppressed(all P＜0.05).Conclusions Piezo1 and YAP1 are synergistically highly expressed in LIHC.Piezo1 enhances the proliferation and invasion abilities of LIHC by regulating YAP1,reshaping tumor microenvironment to promote the malignancy of liver cancer and harm patients'prognosis.关键词
肝细胞癌/肿瘤微环境/细胞外基质/Piezo1/Yes相关转录蛋白1Key words
hepatocellular carcinoma/tumor microenvironment/extracellular matrix/Piezo1/Yes-associated transcription protein 1
