Fu Changhao, Wang Lu, Tian Geer, Zhang Chen, Zhao Yuanyuan, Xu Hao, Su Manman, Wang Yi
Department of Regenerative Medicine, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
Xiamen Institute of Rare Earth Materials, Chinese Academy of Sciences, Xiamen, Fujian 361021, P.R. China.
Oncol Lett. 2019 Feb;17(2):1798-1806. doi: 10.3892/ol.2018.9796. Epub 2018 Dec 5.
Oncostatin M (OSM) induces the differentiation of liver cancer stem cells (LCSCs) and increases sensitivity to the chemotherapeutic agent 5-fluorouracil, whereas salinomycin (Sal) induces apoptosis in cancer stem cells and inhibits the proliferation of liver cancer cells. However, there have been no studies investigating the anticancer effects of combination treatment with OSM and Sal. In the present study, we investigated the synergistic effects of OSM and Sal on LCSCs, the CD133 subpopulations from HepG2 human liver cancer cells. CD133 LCSCs were isolated using an immunomagnetic bead technique and identified through colony formation. After incubating with OSM and Sal, the ability of LCSC proliferation and invasion, as well as apoptosis rates were evaluated, and the expression of stemness-related genes was examined by quantitative real-time polymerase chain reaction. Additionally, the secretion of α-fetoprotein (AFP) and albumin (ALB) were analyzed by enzyme-linked immunosorbent assay. Our results indicated that OSM combined with Sal significantly suppressed LCSC proliferation and invasion and induced apoptosis, as determined by flow cytometry and increases in cleaved caspase-3 levels detected by western blotting. The results of the JC-1 staining assay indicated that this effect involved the mitochondrial pathway. Moreover, combination treatment reduced the expression of CD133 in LCSCs and suppressed stemness-related gene expression. Furthermore, the LCSCs produced lower levels of AFP and higher levels of ALB following combination treatment. In all experiments, combination treatment elicited more efficient anticancer effects on LCSCs as compared with single-drug treatment; therefore, our results demonstrated that combined treatment with OSM and Sal inhibited proliferation and induced differentiation and apoptosis in LCSCs, suggesting combined use of OSM and Sal as a therapeutic strategy for liver cancer.
抑瘤素M（OSM）可诱导肝癌干细胞（LCSCs）分化，并增加其对化疗药物5-氟尿嘧啶的敏感性，而沙林霉素（Sal）可诱导癌症干细胞凋亡并抑制肝癌细胞增殖。然而，尚未有研究探讨OSM与Sal联合治疗的抗癌效果。在本研究中，我们研究了OSM和Sal对LCSCs（来自HepG2人肝癌细胞的CD133亚群）的协同作用。使用免疫磁珠技术分离CD133 LCSCs，并通过集落形成进行鉴定。在用OSM和Sal孵育后，评估LCSC增殖和侵袭能力以及凋亡率，并通过定量实时聚合酶链反应检测干性相关基因的表达。此外，通过酶联免疫吸附测定分析甲胎蛋白（AFP）和白蛋白（ALB）的分泌情况。我们的结果表明，OSM与Sal联合使用可显著抑制LCSC增殖和侵袭并诱导凋亡，这通过流式细胞术确定，并且通过蛋白质印迹法检测到裂解的半胱天冬酶-3水平升高得以证实。JC-1染色试验结果表明，这种作用涉及线粒体途径。此外，联合治疗降低了LCSCs中CD133的表达并抑制了干性相关基因的表达。此外，联合治疗后LCSCs产生的AFP水平较低，ALB水平较高。在所有实验中，与单药治疗相比，联合治疗对LCSCs产生了更有效的抗癌作用；因此，我们的结果表明，OSM与Sal联合治疗可抑制LCSCs的增殖并诱导其分化和凋亡，提示OSM与Sal联合使用可作为肝癌的一种治疗策略。
