JS-K, a nitric oxide prodrug, induces DNA damage and apoptosis in HBV-positive hepatocellular carcinoma HepG2.2.15 cell
JS-K, a nitric oxide prodrug, induces DNA damage and apoptosis in HBV-positive hepatocellular carcinoma HepG2.2.15 cell
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JS-K，一种一氧化氮前药，诱导 HBV 阳性肝细胞癌 HepG2.2.15 细胞 DNA 损伤和细胞凋亡
DOI：
10.1016/j.biopha.2017.05.141
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发表时间：
2017-08-01
影响因子：
7.5
通讯作者：
Wang, Huan
中科院分区：
文献类型：
--
作者：
Liu, Zhengyun;Li, Guangmin;Wang, Huan
肝细胞癌是导致癌症相关死亡的最主要原因，而85%的肝细胞癌是由慢性乙肝病毒感染引起的，患者的预后和HBVDNA水平的降低仍不尽如人意。JS-K是一种释放一氧化氮的二氮杂二酸盐，对多种肿瘤都有疗效，但对乙肝病毒阳性的肝癌的治疗效果却知之甚少。我们发现JS-K降低了HBV2.2.15阳性细胞中HBs Ag和HBeAg的表达。本研究旨在进一步检测JS-K对人肝癌细胞株HepG2.2.15的抗肿瘤作用。采用四甲基偶氮唑盐比色法和集落形成实验研究JS-K对细胞生长的抑制作用，采用划痕实验和Transwell实验检测细胞迁移情况。流式细胞仪检测细胞周期。采用免疫荧光、流式细胞仪、免疫印迹等方法检测细胞DNA损伤和细胞凋亡。JS-K以剂量依赖方式抑制HepG2.2.15细胞生长，抑制细胞集落形成和迁移，使细胞聚集于G2期。JS-K(1-20 mU M)可增加DNA损伤相关蛋白磷酸化H(2)AX(Gamma H(2)AX)、检查点蛋白1的磷酸化(p-Chk1)、检查点蛋白2的磷酸化(p-Chk2)、共济失调性毛细血管扩张突变(ATM)、共济失调性毛细血管扩张相关突变的Rad3(p-ATR)和凋亡相关蛋白裂解caspase-3、裂解caspase-7、裂解多聚ADP-核糖聚合酶(裂解PARP)的表达。本研究证实JS-K对乙肝病毒阳性的HepG2.2.15细胞具有抗病毒作用，其作用机制不仅与抑制HBs Ag和HBeAg的分泌有关，还与诱导DNA损伤和细胞凋亡有关。JS-K是一种很有前途的抗癌候选药物，可用于治疗乙肝病毒阳性的肝癌。(C)2017年爱思唯尔·马森公司。版权所有。
Hepatocellular carcinoma (HCC) is the most important cause of cancer-related death, and 85% of HCC is caused by chronic HBV infection, the prognosis of patients and the reduction of HBV DNA levels remain unsatisfactory. JS-K, a nitric oxide-releasing diazeniumdiolates, is effective against various tumors, but little is known on its effects on HBV positive HCC. We found that JS-K reduced the expression of HBsAg and HBeAg in HBV-positive HepG2.2.15 cells. This study aimed to further examine anti-tumor effects of JS-K on HepG2.2.15 cells. The MTT assay and colony forming assay were used to study the cell growth inhibition of JS-K; scratch assay and transwell assay were performed to detect cell migration. The cell cycle was detected by flow cytometry. The immunofluorescence, flow cytometry analysis, and western blot were used to study DNA damage and cell apoptosis. JS-K inhibited HepG2.2.15 cell growth in a dose-dependent manner, suppressed cell colony formation and migration, arrested cells gather in the G2 phase. JS-K (1-20 mu M) increased the expression of DNA damage-associated protein phosphorylation H(2)AX (gamma H(2)AX), phosphorylation of checkpoint kinase 1 (p-Chk1), phosphorylation of checkpoint kinase 2 (p-Chk2), ataxia-telangiectasia mutated (ATM), phosphorylation of ataxia-telangiectasia mutated rad3-related (p-ATR) and apoptotic-associated proteins cleaved caspase-3, cleaved caspase-7, cleaved poly ADP-ribose polymerase (cleaved PARP). The study demonstrated JS-K is effective against HBV-positive HepG2.2.15 cells, the mechanisms are not only related to inhibition of HBsAg and HBeAg secretion, but also related with induction of DNA damage and apoptosis. JS-K is a promising anti-cancer candidate against HBV-positive HCC. (C) 2017 Elsevier Masson SAS. All rights reserved.