Shi Lu, Zhou Shan-Shan, Chen Wan-Bo, Xu Lei
Hubei University of Chinese Medicine, Ministry of Education Key Laboratory of Traditional Chinese Medicine Resources and Compounds, Wuhan, Hubei 430065, P.R. China.
Guangzhou University of Chinese Medicine, The First Clinical Medical College of Guangzhou University of TCM, Guangzhou, Guangdong 510405, P.R. China.
Exp Ther Med. 2017 Jun;13(6):3116-3122. doi: 10.3892/etm.2017.4314. Epub 2017 Apr 6.
Hepatocellular carcinoma (HCC) is the second leading cause of cancer mortality in China and the third leading cause of cancer mortality worldwide. The mechanisms involved in the development and progression of HCC are not well understood. In the present study, the functions of endothelin-1 (ET-1) in HCC were studied and its underlying mechanisms were investigated. ET-1, B-cell lymphoma 2 (Bcl-2), Bcl-2 related protein 4 (Bax), matrix metalloproteinase (MMP)-2 and MMP-9 expression was measured by reverse transcription-quantitative polymerase chain reaction and western blotting. Cell proliferation was measured via Cell Counting kit-8 assay. Flow cytometry was performed for cell cycle and apoptosis analysis. Migration was measured via Transwell assay. The results demonstrated that ET-1 expression significantly increased in HCC tissues compared with the normal tissues of patients in The Cancer Genome Atlas dataset (P<0.01). Furthermore, downregulation of ET-1 was able to significantly inhibit cell proliferation and growth (P<0.01) and (P<0.01), and induce cell cycle arrest (P<0.05) and apoptosis (P<0.01) in the HCC SMMC-7721 cell line. Bioinformatics analysis demonstrated that the cell apoptosis signaling pathway was activated by ET-1. The ratio of B-cell lymphoma (Bcl-2) -related protein 4 (Bax)/Bcl-2 was significantly increased by downregulation of ET-1 (P<0.01). ET-1 downregulation also inhibited migration of SMMC-7721 cells (P<0.05) via decreasing levels of matrix metalloproteinase (MMP) -2 (P<0.05) and MMP-9 (P>0.05). These results suggest that ET-1 may be able to affect the apoptosis and migration of HCC cells via modulation of the Bax/Bcl-2 ratio and expression levels of MMP-2 and MMP-9, which indicates that ET-1 maybe a potential novel target for HCC treatment.
肝细胞癌（HCC）是中国癌症死亡的第二大原因，也是全球癌症死亡的第三大原因。HCC发生和发展所涉及的机制尚未完全明确。在本研究中，对内皮素-1（ET-1）在HCC中的功能进行了研究，并探究了其潜在机制。通过逆转录-定量聚合酶链反应和蛋白质印迹法检测ET-1、B细胞淋巴瘤2（Bcl-2）、Bcl-2相关蛋白4（Bax）、基质金属蛋白酶（MMP）-2和MMP-9的表达。通过细胞计数试剂盒-8法检测细胞增殖。进行流式细胞术分析细胞周期和凋亡。通过Transwell法检测迁移。结果表明，与癌症基因组图谱数据集中患者的正常组织相比，HCC组织中ET-1表达显著增加（P<0.01）。此外，ET-1的下调能够显著抑制HCC SMMC-7721细胞系中的细胞增殖和生长（P<0.01），并诱导细胞周期停滞（P<0.05）和凋亡（P<0.01）。生物信息学分析表明，细胞凋亡信号通路被ET-1激活。ET-1的下调显著增加了B细胞淋巴瘤（Bcl-2）相关蛋白4（Bax）/Bcl-2的比值（P<0.01）。ET-1的下调还通过降低基质金属蛋白酶（MMP）-2（P<0.05）和MMP-9（P>0.05）的水平抑制了SMMC-7721细胞的迁移（P<0.05）。这些结果表明，ET-1可能通过调节Bax/Bcl-2比值以及MMP-2和MMP-9的表达水平来影响HCC细胞的凋亡和迁移，这表明ET-1可能是HCC治疗的一个潜在新靶点。
