In vivo study revealed pro-tumorigenic effect of CMTM3 in hepatocellular carcinoma involving the regulation of peroxisome proliferator-activated receptor gamma (PPARγ)
In vivo study revealed pro-tumorigenic effect of CMTM3 in hepatocellular carcinoma involving the regulation of peroxisome proliferator-activated receptor gamma (PPARγ)
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DOI：
10.1007/s13402-022-00733-1
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发表时间：
2022-10-26
影响因子：
6.6
通讯作者：
Liu，Fujun
中科院分区：
文献类型：
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作者：
Wang，Jiahui;Chu，Hongjin;Liu，Fujun
目的澄清CMTM3在肝细胞癌（HCC）发生发展过程中功能的模糊性，探讨其分子机制。方法采用CRISPR-Cas9技术建立小鼠cmm3 - ko C57BL/6菌株。腹腔注射100、25 mg/kg急性肝损伤模型和肝细胞癌模型。BW n -亚硝基二乙胺（DEN）对雄性小鼠。采用血清谷丙转氨酶、谷丙转氨酶水平及HE染色评价肝功能及组织学。采用RNA-seq、RT-qPCR、Western blotting、免疫组织化学、免疫荧光等检测肝组织中基因和蛋白的表达。通过STRING和拓扑测量研究蛋白-蛋白相互作用。使用UALCAN数据库分析CMTM3和PPARs mRNA表达与患者生存率。结果成功证实了KO小鼠cmtm3的全基因敲除。cmtm3敲除减轻了den诱导的急性肝组织完整性和肝功能损伤，减少了DNA损伤和凋亡，并显著减少了细胞数量（WT: 8.7±5.5vs）。KO: 2.7±3.1，P= 0.0394)，肿瘤总大小（WT: 130.9±181.8 mm2vs）。KO: 9.3±11.5 mm2，P= 0.026)。在机制上，cmtm3敲除导致DEN中毒后Pparγ及其下游脂质代谢基因（如Adipoq）的表达减少和失活。CMTM3和PPARγ在HCC中均过表达，这两个基因的高水平与HCC患者的总生存率较差相关。本研究明确了CMTM3在HCCin体内的促肿瘤作用，可能是通过上调PPARγ和激活PPAR通路实现的。图形抽象
PurposeTo clarify the ambiguity of the function of CMTM3 in the development of hepatocellular carcinoma (HCC) and explore its molecular mechanism.MethodsTheCmtm3-KO C57BL/6 mouse strain was established using CRISPR-Cas9. Acute liver damage and HCC models were induced by peritoneal injection of 100 or 25 mg/kg.BW N-Nitrosodiethylamine (DEN) to male mice. Liver function and histology were evaluated by blood serum levels of AST and ALT, and HE staining. Gene and protein expression in liver tissues was investigated by RNA-seq, RT-qPCR, Western blotting, immunohistochemistry, and immunofluorescence. Protein–protein interactions were studied by STRING and topological measures. The mRNA expression of CMTM3 and PPARs and patient survival were analyzed using the UALCAN database.ResultsGlobal knockout ofCmtm3in KO mice was successfully confirmed.Cmtm3knockout alleviated DEN-induced acute damage to liver histological integrity and liver function, reduced DNA damage and apoptosis, and also caused a significantly reduced number (WT: 8.7 ± 5.5vs. KO: 2.7 ± 3.1,P= 0.0394) and total size of tumors (WT: 130.9 ± 181.8 mm2vs. KO: 9.3 ± 11.5 mm2,P= 0.026) in the liver. Mechanistically,Cmtm3knockout resulted in reduced expression and inactivation of Pparγ and its downstream lipid metabolism genes (e.g. Adipoq) upon DEN intoxication. CMTM3 and PPARγ were both overexpressed in HCC, and higher levels of both genes were associated with worse overall survival of HCC patients.ConclusionThis study clarified the pro-tumorigenesis role of CMTM3 in HCCin vivo, possibly through the upregulation of PPARγ and activation of the PPAR pathway.Graphical abstract