Huang Chuan, Wang Si-Hong, Liu Tao-Tao, Wu Mei-Yi, Cai Kai-Ning, Xie Zhan-Hong, Zhao Rui-Qiang, Wen Yan
Department of Pharmacy, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China.
Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Guangxi Medical University, Nanning, 530021, China.
Recent Pat Anticancer Drug Discov. 2024 Jul 11. doi: 10.2174/0115748928293750240619092747.
The emergence of drug resistance to oxaliplatin (OXA) is one of the critical obstacles in the therapy of advanced Hepatocellular Carcinoma (HCC). As an ethyl derivative of the natural compound epigallocatechin gallate (epigallocatechin-3-gallate, EGCG), Y6 was found to be able to enhance the sensitivity of HCC cells to doxorubicin. This study aimed to investigate the effect of Y6 on oxaliplatin resistance in HCC.
MTT was used to determine the reversal effect of Y6 on OXA resistance. To further explore the reversal mechanism, we treated OXA alone or in combination with Y6 or EGCG in drugresistant cells and observed the morphological changes of the cells. At the same time, transwell assay was used to detect the invasion and migration ability of cells. Moreover, Real-time PCR and Western blot analysis were performed to determine the expression levels of the miR-338-3p gene, HIF-1α/Twist proteins, and EMT-related proteins.
We found that Y6 could inhibit the proliferation of HCC cells and effectively reverse the drug resistance of oxaliplatin-resistant human liver cancer cells (SMMC-7721/OXA) to OXA, and the reversal effect was more significant than that of its lead drug EGCG. Most of the cells in the control group and OXA group showed typical mesenchymal-like cell morphology, while most of the cells in co-administration groups showed typical epithelioid cell morphology, and the ability of the cells to invade and migrate decreased dramatically, particularly in Y6 plus OXA group. At the same time, Y6 could up-regulate the EMT epithelial marker protein E-cadherin and down-regulate the interstitial marker protein Vimentin. In addition, in co-administration groups, the expression of miR-338-3p was up-regulated, while the expression of HIF-1α and Twist was down-regulated.
Y6 significantly enhanced the susceptibility of drug-resistant cells to OXA, and the process may be related to the regulation of miR-338-3p/HIF-1α / TWIST pathway to inhibit EMT. Therefore, Y6 could be considered an effective medication resistance reversal agent, which could improve the therapeutic effect for hepatocellular cancer patients.
对奥沙利铂（OXA）产生耐药性是晚期肝细胞癌（HCC）治疗中的关键障碍之一。作为天然化合物表没食子儿茶素没食子酸酯（表没食子儿茶素-3-没食子酸酯，EGCG）的乙基衍生物，Y6被发现能够增强肝癌细胞对阿霉素的敏感性。本研究旨在探讨Y6对肝癌细胞奥沙利铂耐药性的影响。
采用MTT法测定Y6对奥沙利铂耐药的逆转作用。为进一步探究逆转机制，我们在耐药细胞中单独用奥沙利铂处理或联合Y6或EGCG处理，并观察细胞的形态变化。同时，采用Transwell实验检测细胞的侵袭和迁移能力。此外，进行实时荧光定量PCR和蛋白质免疫印迹分析以确定miR-338-3p基因、HIF-1α/Twist蛋白以及上皮-间质转化（EMT）相关蛋白的表达水平。
我们发现Y6能够抑制肝癌细胞的增殖，并有效逆转耐奥沙利铂的人肝癌细胞（SMMC-7721/OXA）对奥沙利铂的耐药性，且逆转作用比其先导药物EGCG更显著。对照组和奥沙利铂组的大多数细胞呈现典型的间充质样细胞形态，而联合用药组的大多数细胞呈现典型的上皮样细胞形态，并且细胞的侵袭和迁移能力显著下降，尤其是在Y6加奥沙利铂组。同时，Y6能够上调EMT上皮标志物蛋白E-钙黏蛋白并下调间质标志物蛋白波形蛋白。此外，在联合用药组中，miR-338-3p的表达上调，而HIF-1α和Twist的表达下调。
Y6显著增强了耐药细胞对奥沙利铂的敏感性，这一过程可能与miR-338-3p/HIF-1α/TWIST通路调控抑制EMT有关。因此，Y6可被视为一种有效的耐药逆转剂，有望提高肝癌患者的治疗效果。
