Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China
Department of Gastroenterology, The First Affiliated Hospital, Gannan Medical University, Ganzhou 341000, China.
Biosci Rep. 2018 Nov 9;38(6). doi: 10.1042/BSR20180854. Print 2018 Dec 21.
To explore whether aspirin (ASA) enhances the sensitivity of hepatocellular carcinoma (HCC) side population (SP) cells to doxorubicin (Doxo) via miR-491/ATP-binding cassette sub-family G member 2 (ABCG2). Non-SP and SP cells were isolated from MHCC-97L cell line using flow cytometry analysis and fluorescence-activated cell sorting. Colony formation assay was performed to determine the colony-formation ability of cells. Cell viability of SP cells was determined with the MTT assay. Luciferase reporter assay was applied in confirming the binding between miR-491 and ABCG2. Although the Doxo treatment lowered the colony-formation ability of both non-SP and SP cells, the colony-formation ability of SP cells was 2-fold higher than that of non-SP cells (<0.05). Doxo slightly inhibited the cell viability of SP cells in a concentration-dependent manner; the addition of ASA dramatically enhanced the inhibitory effect of Doxo on SP cell viability in a concentration-dependent manner (<0.05). Compared with non-SP cells, the miR-491 expression was significantly decreased in SP cells, which was significantly reversed by ASA (<0.05). miR-491 directly controlled the ABCG2 expression. In the presence of Doxo, miR-491 inhibitor reduced the inhibitory effect of ASA on the cell viability of SP cells, which was significantly reversed by knockdown of ABCG2 (<0.05). ASA enhanced the sensitivity of SP cells to Doxo via regulating the miR-491/ABCG2 signaling pathway.
探讨阿司匹林（ASA）是否通过 miR-491/ATP 结合盒亚家族 G 成员 2（ABCG2）增强肝癌（HCC）侧群（SP）细胞对阿霉素（Doxo）的敏感性。使用流式细胞术分析和荧光激活细胞分选从 MHCC-97L 细胞系中分离非 SP 和 SP 细胞。集落形成实验用于确定细胞的集落形成能力。MTT 测定法用于测定 SP 细胞的细胞活力。荧光素酶报告测定法用于确认 miR-491 与 ABCG2 之间的结合。虽然 Doxo 处理降低了非 SP 和 SP 细胞的集落形成能力，但 SP 细胞的集落形成能力是非 SP 细胞的 2 倍（<0.05）。Doxo 以浓度依赖性方式轻微抑制 SP 细胞的细胞活力；ASA 的加入以浓度依赖性方式显着增强 Doxo 对 SP 细胞活力的抑制作用（<0.05）。与非 SP 细胞相比，SP 细胞中的 miR-491 表达明显降低，ASA 明显逆转（<0.05）。miR-491 直接控制 ABCG2 的表达。在 Doxo 存在的情况下，miR-491 抑制剂降低了 ASA 对 SP 细胞活力的抑制作用，ABCG2 的敲低明显逆转（<0.05）。ASA 通过调节 miR-491/ABCG2 信号通路增强了 SP 细胞对 Doxo 的敏感性。
