Choi Sung Hoon, Cho Kyung Joo, Yun Sung Ho, Jin Bora, Lee Ha Young, Ro Simon W, Kim Do Young, Ahn Sang Hoon, Han Kwang-Hyub, Park Jun Yong
Yonsei Liver Center, Yonsei University College of Medicine, Seoul, Republic of Korea.
BK21 plus project for medical science, Yonsei University College of Medicine, Seoul, Republic of Korea.
J Cancer. 2020 Feb 10;11(9):2442-2452. doi: 10.7150/jca.39380. eCollection 2020.
Hepatocellular carcinoma is a malignant disease with improved hepatic regeneration and survival, and is activated by human telomere transferase (hTERT). hTERT is expressed during early fetal development and switched off in most adult tissues, but it becomes reactivated in HCC. The exact mechanism regulating these expression changes remains unknown during HCC progress. We evaluated the relationship between hTERT expression and human kruppel-related 3 (HKR3) and cell cycle-related factors in HCC cell lines. Following transfection for hTERT knockdown and HKR3 overexpression, proteomic and transcriptomic analyses related to hTERT were performed using liquid chromatography/mass spectrometry (LC/MS) and RNA sequencing (RNAseq) in HCC cell lines. The expression levels of hTERT, HKR3, and cell cycle-related factors were measured using western blotting, and tumor growth were evaluated via cell proliferation and cell cycle assays. Transcriptomic and proteomic analyses showed that HKR3, hTERT and cyclin-dependent kinase inhibitor 2A (CDKN2A) were correlated. Up-regulation of HKR3 expression decreased hTERT and cyclin activation and suppressed the G1/S phase of the cell cycle through CDKN2A activation. Our results suggest that HKR3 induced regulation of cell cycle through hTERT inhibition and CDKN2A activation. Our results will facilitate further exploration of the pathways regulating human telomerase activity in HCC cell lines.
肝细胞癌是一种具有改善的肝再生和生存率的恶性疾病，由人端粒转移酶（hTERT）激活。hTERT在胎儿早期发育过程中表达，在大多数成人组织中关闭，但在肝癌中重新激活。在肝癌进展过程中，调节这些表达变化的确切机制仍不清楚。我们评估了hTERT表达与人克鲁ppel相关3（HKR3）和肝癌细胞系中细胞周期相关因子之间的关系。在转染hTERT敲低和HKR3过表达后，使用液相色谱/质谱（LC/MS）和RNA测序（RNAseq）在肝癌细胞系中进行了与hTERT相关的蛋白质组学和转录组学分析。使用蛋白质印迹法测量hTERT、HKR3和细胞周期相关因子的表达水平，并通过细胞增殖和细胞周期测定评估肿瘤生长。转录组学和蛋白质组学分析表明，HKR3、hTERT和细胞周期蛋白依赖性激酶抑制剂2A（CDKN2A）相关。HKR3表达上调降低了hTERT和细胞周期蛋白的激活，并通过激活CDKN2A抑制细胞周期的G1/S期。我们的结果表明，HKR3通过抑制hTERT和激活CDKN2A诱导细胞周期调控。我们的结果将有助于进一步探索肝癌细胞系中调节人端粒酶活性的途径。
