Wang Shanshan S, Chen Yuhan H, Chen Ning, Wang Lijun J, Chen Dexi X, Weng Honglei L, Dooley Steven, Ding Huiguo G
Department of Gastrointestinal and Hepatology, Beijing You' An Hospital Affiliated to Capital Medical University, Beijing, 100069, China.
Cell Biology Laboratory, Beijing Institute of Hepatology, Beijing, 100069, China.
Cell Death Dis. 2017 Mar 23;8(3):e2688. doi: 10.1038/cddis.2017.18.
Hydrogen sulfide (HS), in its gaseous form, plays an important role in tumor carcinogenesis. This study investigated the effects of HS on the cell biological functions of hepatocellular carcinoma (HCC). HCC cell lines, HepG2 and HLE, were treated with NaHS, a donor of HS, and rapamycin, a classic autophagy inducer, for different lengths of time. Western blotting, immunofluorescence, transmission electron microscopy (TEM), scratch assay, CCK-8 and flow cytometric analysis were carried out to examine the effects of HS on HCC autophagy, cell behavior and PI3K/Akt/mTOR signaling. Treatment with NaHS upregulated expression of LC3-II and Atg5, two autophagy-related proteins, in HepG2 and HLE cells. TEM revealed increased numbers of intracellular double-membrane vesicles in those cells treated with NaHS. Like rapamycin, NaHS also significantly inhibited expression of p-PI3K, p-Akt and mTOR proteins in HCC cells. Interestingly, the expression of LC3-II was further increased when the cells were treated with NaHS together with rapamycin. In addition, NaHS inhibited HCC cell migration, proliferation and cell division. These findings show that HS can induce HCC cell apoptosis. The biological function of the gasotransmitter HS in HCC cells was enhanced by the addition of rapamycin. Hydrogen sulfide influences multiple biological functions of HCC cells through inhibiting the PI3K/Akt/mTOR signaling pathway.
气态硫化氢（HS）在肿瘤发生过程中发挥着重要作用。本研究调查了HS对肝癌（HCC）细胞生物学功能的影响。使用HS供体硫氢化钠（NaHS）和经典自噬诱导剂雷帕霉素对肝癌细胞系HepG2和HLE进行不同时长的处理。采用蛋白质免疫印迹法、免疫荧光法、透射电子显微镜（TEM）、划痕试验、CCK-8法和流式细胞术分析来检测HS对肝癌细胞自噬、细胞行为及PI3K/Akt/mTOR信号通路的影响。用NaHS处理可上调HepG2和HLE细胞中两种自噬相关蛋白LC3-II和Atg5的表达。TEM显示，经NaHS处理的细胞内双膜囊泡数量增加。与雷帕霉素一样，NaHS也显著抑制肝癌细胞中p-PI3K、p-Akt和mTOR蛋白的表达。有趣的是，当细胞同时用NaHS和雷帕霉素处理时，LC3-II的表达进一步增加。此外，NaHS抑制肝癌细胞迁移、增殖和细胞分裂。这些发现表明，HS可诱导肝癌细胞凋亡。添加雷帕霉素可增强气体信号分子HS在肝癌细胞中的生物学功能。硫化氢通过抑制PI3K/Akt/mTOR信号通路影响肝癌细胞的多种生物学功能。
