关键词: 肝细胞癌, ADH1B, 脉管侵犯, 增殖, 侵袭, 迁移, 预后

Abstract: Objective To investigate the clinical value of alcohol dehydrogenase 1B (ADH1B)in patients with hepatocellular carcinoma with vascular invasion (VI⁃HCC) and its effect on the biological function of liver cancer cells. Methods RNA sequencing of VI⁃HCC tissues, and GO, KEGG, GSEA, WGCNA analysis were performed. The expression level of ADH1B protein in tissues was detected by immunohistochemistry. The Kaplan⁃Meier method was used to draw the survival curve and the log⁃rank test was performed to analyze the survival difference of patients with different ADH1B protein expression groups. Univariate and multivariate Cox regression models were used to analyze the independent risk factors affecting the prognosis of patients with VI⁃HCC after surgery, and Spearman test was used to analyze the correlation between ADH1B and AFP/Ki67. The lentivirus overexpressing ADH1B and its negative control were used to infect the Huh7 cells to construct a stable ADH1B overexpression strain. The expression level of ADH1B mRNA in cells were detected by qRT⁃PCR. EDU and plate cloning assay were used to detect the proliferation ability of the cells. Transwell invasion and migration assay and scratch assay were used to detect the invasion and migration ability of the cells. Flow cytometry was used to detect apoptosis of cells. Results The results of RNA sequencing showed that the primary tumors of VI⁃HCC patients with poor prognosis after radical resection was characterized by metabolic reprogramming. The results of immunohistochemistry showed that the expression level of ADH1B protein in VI⁃HCC tissues was significantly lower than that in adjacent normal tissues (P<0.001). The expression of ADH1B protein in patients with good prognosis was significantly higher than that in patients with poor prognosis (P<0.05). Low expression of ADH1B could reduce both the relapse⁃free survival (RFS) and the overall survival (OS) of patients (all P<0.01). The expression level of ADH1B was negatively correlated with AFP and Ki67 levels (all P<0.01). Cox regression model showed that low expression level of ADH1B was an independent risk factor for postoperative RFS and OS in the patients with VI⁃HCC. Cytological results showed that overexpression of ADH1B could inhibit the proliferation, invasion and migration of cells, and promoted the apoptosis (all P<0.05). Conclusions The expression of liver metabolism⁃detoxification gene ADH1B is down⁃regulated in HCC, and its low expression reduces the RFS and OS in patients with VI⁃HCC. The possible mechanism is that ADH1B inhibits the proliferation, invasion and migration of HCC cells, and promotes the apoptosis of HCC cells.

Key words: Hepatocellular carcinoma, ADH1B, Vascular invasion, Proliferation, Invasion, Migration, Prognosis