背景:奥沙利铂(OXA)耐药性的出现是晚期肝细胞癌(HCC)治疗的关键障碍之一。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),发现Y6能够增强HCC细胞对阿霉素的敏感性。本研究旨在探讨Y6对HCC奥沙利铂耐药的影响。
方法:MTT用于确定Y6对OXA抗性的逆转作用。为了进一步探索逆转机制,我们在耐药细胞中单独或与Y6或EGCG联合处理OXA,并观察细胞的形态变化。同时,transwell法检测细胞的侵袭和迁移能力。此外,进行实时PCR和Western印迹分析以确定miR-338-3p基因的表达水平。HIF-1α/Twist蛋白,和EMT相关蛋白.
结果:我们发现Y6可以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OXA的耐药性,且逆转效应比其先导药物EGCG更为显著。对照组和OXA组大部分细胞呈典型的间充质样细胞形态,而共同给药组中的大多数细胞表现出典型的上皮样细胞形态,细胞侵入和迁移的能力急剧下降,特别是Y6加OXA组。同时,Y6可以上调EMT上皮标记蛋白E-cadherin,下调间质标记蛋白Vimentin。此外,在共同管理团体中,miR-338-3p的表达上调,而HIF-1α和Twist的表达下调。
结论:Y6能显著增强耐药细胞对OXA的敏感性,此过程可能与调控miR-338-3p/HIF-1α/TWIST通路抑制EMT有关。因此,Y6可以被认为是一种有效的药物耐药逆转剂,提高肝癌患者的治疗效果。
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.