Uncropped western blots.

V-ATPase Subunit G1 (ATP6V1G1) is one of the subunits of Vacuolar ATPases. Previous studies have indicated that ATP6V1G1 plays important roles in hepatocellular carcinoma (HCC) and is associated with HCC progression. However, the effect of ATP6V1G1 in HCC requires further elucidation. The aim of the present study was to explore the roles of ATP6V1G1 in HCC and further decipher the detailed mechanism. To identify the phosphorylated proteins regulated by ATP6V1G1 in HCC, phosphoproteomics and LC-MS/MS analysis were performed on HepG2 with overexpression of ATP6V1G1 and empty vector. Western blotting was applied to validate the differentially expressed phosphorylated proteins (DEPPs). As a result, 163 DEPPs were identified by proteomics; two up-regulated phosphorylated proteins (p-RPS6(Ser235)) and (p-SQSTM1(Ser272)) and two down-regulated phosphorylated proteins (p-PDPK1(Ser241)) and (p-EEF2 (Ser57)) were validated. Taken together, this study highlighted the potential impact of ATP6V1G1 on tumor progression, which may be beneficial to liver cancer related basic research.V型ATP酶亚基G1（V-ATPase Subunit G1, ATP6V1G1）是液泡型ATP酶的亚基之一。既往研究表明，ATP6V1G1在肝细胞癌（hepatocellular carcinoma, HCC）中发挥重要作用，且与肝细胞癌的进展密切相关。然而，ATP6V1G1在肝细胞癌中的具体调控效应仍有待进一步阐明。本研究旨在探讨ATP6V1G1在肝细胞癌中的作用，并进一步解析其潜在的分子机制。为筛选ATP6V1G1在肝细胞癌中调控的磷酸化蛋白，本研究对过表达ATP6V1G1的肝癌细胞系HepG2及转染空载体的对照组细胞开展了磷酸化蛋白质组学（phosphoproteomics）分析与液相色谱-串联质谱（LC-MS/MS）检测。随后采用蛋白质免疫印迹（Western blotting）对筛选得到的差异表达磷酸化蛋白（differentially expressed phosphorylated proteins, DEPPs）进行验证。实验结果显示，通过蛋白质组学共鉴定出163个差异表达磷酸化蛋白；经验证，上调的磷酸化蛋白为p-RPS6(Ser235)与p-SQSTM1(Ser272)，下调的磷酸化蛋白为p-PDPK1(Ser241)与p-EEF2(Ser57)。综上，本研究揭示了ATP6V1G1对肿瘤进展的潜在影响，可为肝癌相关基础研究提供有益参考。