miR-199a-5p/GPR89A axis modulates sorafenib resistance in hepatocellular carcinoma through glutamate metabolic reprogramming

作者：Yao Li, Cai Yi, Guangliang Duan, Yulin Du, Junjie Hu, Xue Han, Xiaolong Tang, Liangxian Jiang, Junqi Ren, Qi Huo · 发表于：Science Progress · 年份：2025 · DOI：10.1177/00368504251381592 · 被引用次数：2 · 研究领域：Cancer, Hypoxia, and Metabolism、Cancer, Stress, Anesthesia, and Immune Response、PI3K/AKT/mTOR signaling in cancer

ObjectiveSorafenib is a key targeted therapeutic agent for hepatocellular carcinoma (HCC). However, the emergence of drug resistance greatly limits its clinical efficacy. We found that GPR89A is markedly overexpressed in both sorafenib-resistant HCC cell lines and clinical tumor specimens. Nevertheless, the exact biological role of GPR89A in sorafenib-resistant HCC remains to be elucidated.MethodsTo generate sorafenib-resistant HCC cell lines, cells were cultured in medium with 10 μM sorafenib. GPR89A expression in these cells was examined by quantitative reverse transcription-polymerase chain reaction and Western blot. The impact of GPR89A inhibition on cell proliferation was assessed using cell counting kit-8 and colony formation assays. Bioinformatics predicted miR-199a-5p binding sites in the GPR89A promoter region; luciferase reporter assays were conducted with wild-type or mutant promoter sequences. Glutamate in culture supernatants was measured by enzyme-linked immunosorbent assay, and exogenous glutamate showed dose-dependent inhibition of resistant cell proliferation (MTT assay). Clinical correlation analysis involved 30 pairs of advanced HCC samples. Pearson analysis revealed significant inverse correlations among miR-199a-5p, GPR89A, and metabotropic glutamate receptor 1 (mGluR1) expression.ResultsWe found that the cell surface transmembrane protein GPR89A was substantially overexpressed in both sorafenib-resistant HCC cell lines and clinical tumor specimens. Impor...