Mao Xiaorong, Du Senrong, Yang Zhongxia, Zhang Liting, Peng Xuebin, Jiang Ni, Zhou Haiyu
Department of Infectious Disease, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
Department of Infectious Disease, Lanzhou Pulmonary Hospital, Lanzhou, Gansu 730046, P.R. China.
Mol Med Rep. 2017 Jul;16(1):208-214. doi: 10.3892/mmr.2017.6568. Epub 2017 May 10.
It has been confirmed that the inhibitors of poly ADP-ribose polymerF(^9ase‑1 (PARP‑1) can inhibit the proliferation, apoptosis and invasion of tumor cells. However, the effects of inhibitors of PARP‑1 on hepatocellular carcinoma remain to be elucidated. The aim of the present study was to investigate the effect of three types of PARP‑1 inhibitor on the proliferation, apoptosis and migration of hepatocellular carcinoma in vitro. An MTT assay was performed to detect the proliferation of HepG2 cells following treatment with the PARP‑1 inhibitors, AG014699, BSI‑201 and AZD‑2281. Flow cytometry was used to detect the apoptosis of HepG2 cells, Western blot analysis was used to detect the protein expression of Casepase‑3, Casepase‑8, B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax), Bcl‑2, phosphatase and tensin homolog (PTEN), tissue inhibitor of metalloproteinase (TIMP) 3 and matrix metalloprotease (MMP) 3. A Transwell assay was performed to detect the migration of HepG2 cells. The results showed that AG014699, BSI‑201 and AZD‑2281 had an inhibitory effect on the proliferation of HepG2 cells in a time‑ and concentration‑dependent manner. AG014699 at concentrations of 10, 30 and 50 µmol/l, and BSI‑201 at concentrations of 20, 40 and 60 µmol/l induced the apoptosis of HepG2 cells, and the apoptotic rates were particularly high at 48 h (31, vs. 0.01%; P<0.01 and 24.12, vs. 0.03%, respectively; P<0.01). The protein expression levels of Caspase 3, Caspase 8, Bax, PTEN and TIMP 3 increased with increasing drug concentrations, whereas the protein levels of Bcl‑2 and MMP3 decreased with increasing drug concentrations, and were significantly different compared with those in the control group (P<0.01). In conclusion, AG014699, BSI‑201 and AZD‑2281 inhibitors of PARP‑1 significantly inhibited the proliferation of HepG2 cells, however, AG014699 and BSI‑201 demonstrated more sensitivity, induced apoptosis and inhibited migration of the hepatocellular carcinoma cells, which may be associated with alterations of the apoptosis signaling pathway and the expression of proteins associated with migration.
已证实聚ADP核糖聚合酶-1（PARP-1）抑制剂可抑制肿瘤细胞的增殖、凋亡和侵袭。然而，PARP-1抑制剂对肝细胞癌的作用仍有待阐明。本研究的目的是探讨三种PARP-1抑制剂对体外培养的肝细胞癌增殖、凋亡和迁移的影响。采用MTT法检测PARP-1抑制剂AG014699、BSI-201和AZD-2281处理后HepG2细胞的增殖情况。采用流式细胞术检测HepG2细胞的凋亡情况，采用蛋白质印迹分析检测半胱天冬酶-3（Caspase-3）、半胱天冬酶-8（Caspase-8）、B细胞淋巴瘤-2（Bcl-2）相关X蛋白（Bax）、Bcl-2、磷酸酶和张力蛋白同源物（PTEN）、金属蛋白酶组织抑制剂（TIMP）3和基质金属蛋白酶（MMP）3的蛋白表达。采用Transwell实验检测HepG2细胞的迁移情况。结果显示，AG014699、BSI-201和AZD-2281对HepG2细胞的增殖具有抑制作用，且呈时间和浓度依赖性。浓度为10、30和50 μmol/L的AG014699以及浓度为20、40和60 μmol/L的BSI-201可诱导HepG2细胞凋亡，在48 h时凋亡率尤其高（分别为31% vs. 0.01%；P<0.01和24.12% vs. 0.03%；P<0.01）。随着药物浓度增加，Caspase 3、Caspase 8、Bax、PTEN和TIMP 3的蛋白表达水平升高，而Bcl-2和MMP3的蛋白水平随药物浓度增加而降低，与对照组相比差异有统计学意义（P<0.01）。综上所述，PARP-1抑制剂AG014699、BSI-201和AZD-2281可显著抑制HepG2细胞的增殖，然而，AG014699和BSI-201表现出更高的敏感性，可诱导肝癌细胞凋亡并抑制其迁移，这可能与凋亡信号通路的改变以及迁移相关蛋白的表达有关。
