Department of Pharmacy, Hunan Cancer Hospital/The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, Hunan, P. R. China.
Translational Medicine Centre, Hunan Cancer Hospital / The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, Hunan, P. R. China.
Chem Biol Drug Des. 2024 Jan;103(1):e14396. doi: 10.1111/cbdd.14396. Epub 2023 Dec 6.
Patients with advanced liver cancer may benefit from 5-fluorouracil (5-FU) therapy. However, most of them eventually faced drug resistance, resulting in a poor prognosis. The present study aims to explore the potential mechanism of let-7g/ABCC10 axis in the regulation of 5-FU resistance in liver cancer cells. Huh-7 cells were used to construct 5-FU resistant Huh-7/4X cells. CCK8, flow cytometry, and TUNEL staining were used to detect the characterization of Huh-7 cells and Huh-7/4X cells. Double luciferase report, PCR, and western blot analyses were used to detect the regulatory effects between let-7g and ABCC10. The levels of biomarkers related to cell cycle progression and apoptosis were detected by western blot assays. The role of let-7g in 5-FU sensitivity of liver cancer cells was evaluated in nude mice. Compared with LX-2 cells, the expression of let-7g was decreased in Hep3B, HepG2, Huh-7, and SK-Hep1 cells, with the lowest expression in Huh-7 cells. The sensitivity of Huh-7 cell to 5-FU was positively correlated with let-7g expression. Transfection of let-7g mimics inhibited the viability of Huh-7/4X cells by prolonging the G1 phase, with the downregulation of ABCC10, PCNA, Cyclin D1, and CDK4. Meanwhile, let-7g promoted apoptosis to increase 5-FU sensitivity of Huh-7/4X by downregulating ABCC10, Bcl-XL as well as upregulating Bax, C-caspase 3, and C-PARP. Dual-luciferase assay further confirmed that let-7g inhibited ABCC10 expression by binding to the ABCC10 3'-UTR region. Furthermore, let-7g increased the sensitivity of Huh-7/4X to 5-FU in vitro and in vivo, which can be reversed by ABCC10 overexpression. In conclusion, let-7g sensitized liver cancer cells to 5-FU by downregulating ABCC10 expression.
晚期肝癌患者可能从氟尿嘧啶（5-FU）治疗中获益。然而，他们中的大多数最终面临药物耐药，导致预后不良。本研究旨在探讨 let-7g/ABCC10 轴在调节肝癌细胞对 5-FU 耐药中的潜在机制。使用 Huh-7 细胞构建 5-FU 耐药 Huh-7/4X 细胞。CCK8、流式细胞术和 TUNEL 染色用于检测 Huh-7 细胞和 Huh-7/4X 细胞的特征。双荧光素酶报告、PCR 和 Western blot 分析用于检测 let-7g 和 ABCC10 之间的调节作用。Western blot 检测细胞周期进展和细胞凋亡相关生物标志物的水平。在裸鼠中评估 let-7g 对肝癌细胞对 5-FU 敏感性的作用。与 LX-2 细胞相比，Hep3B、HepG2、Huh-7 和 SK-Hep1 细胞中 let-7g 的表达降低，而 Huh-7 细胞中的表达最低。Huh-7 细胞对 5-FU 的敏感性与 let-7g 表达呈正相关。转染 let-7g 模拟物通过延长 G1 期抑制 Huh-7/4X 细胞的活力，同时下调 ABCC10、PCNA、Cyclin D1 和 CDK4。同时，let-7g 通过下调 ABCC10、Bcl-XL 以及上调 Bax、C-caspase 3 和 C-PARP 促进凋亡，从而提高 Huh-7/4X 对 5-FU 的敏感性。双荧光素酶测定进一步证实 let-7g 通过结合 ABCC10 3'-UTR 区域抑制 ABCC10 表达。此外，let-7g 在体外和体内增加了 Huh-7/4X 对 5-FU 的敏感性，这可以通过 ABCC10 过表达逆转。总之，let-7g 通过下调 ABCC10 表达使肝癌细胞对 5-FU 敏感。
