Original Article
NVP-TNKS656 inhibits the growth of hepatocellular carcinoma cell lines by regulating Hippo signaling pathway
Qiao Yu, Hu Zhehui, Yu Gang, Duan Beibei, Zhang Shuai, Zhao Yunbo, Zhang Zijin, Li Lin
Published 2020-06-14
Cite as Chin J Geriatr, 2020, 39(6): 700-705. DOI: 10.3760/cma.j.issn.0254-9026.2020.06.022
Abstract
ObjectiveTo investigate the effect of a tankyrase inhibitor NVP-TNKS656 on the growth of hepatocellular carcinoma(HCC)cell lines and the involved molecular mechanisms.
MethodsFive HCC cell lines were treated with 0, 2.5, 5.0, 10.0 μmol/L of NVP-TNKS656.The cell lines of HLE and HLF were selected and divided into four groups: 0.0 μmol/L(control or DMSO), 2.5 μmol/L, 5.0 μmol/L, 10.0 μmol/L of NVP-TNKS656 groups.Cells were cultured for 48 h for subsequent experiments.Crystal violet staining was used to count the number of the newly formed cell clones.Western blotting was used to detect the protein expression levels of Yes-associated protein(YAP), angiomotin-like 1(AMOTL1)and AMOTL2.The real-time qRT-PCR was used to detect the mRNA expression of YAP and its downstream connective tissue growth factor(CTGF)and cysteine-rich 61(Cyr61). Dual luciferase reporter gene was used to detect the luciferase activity of transcriptional enhancer activator domain(TEAD)family.
ResultsNVP-TNKS656 inhibited the growth of 5 HCC cell lines in a dose-dependent manner in HLE, HLF, Huh7, MHCC97-H, and MHCC97-L cell lines.There were significant differences in the newly formed cell clone numbers between control(0 μM of NVP-TNKS656)and each of 2.5 μmol/L, 5.0 μmol/L, 10.0 μmol/L of NVP-TNKS656 groups in a dose-dependent manner( F=90.46, 68.58, 191.8, 114.6 and 201.4, all P<0.05). In HLE and HLF cell lines, NVP-TNKS656 significantly reduced the protein level of YAP in a dose-dependent manner and decreased the YAP target gene CTGF(HLE cells: 1.02±0.02, 0.90±0.03, 0.57±0.02, 0.38±0.03, HLF cells: 0.98±0.03, 0.86±0.02, 0.66±0.02, 0.43±0.01)and Cyr61(HLE cells: 1.00±0.01, 0.86±0.02, 0.74±0.03, 0.44±0.03 and HLF cells: 0.99± 0.02, 0.87±0.01, 0.72±0.02, 0.54±0.01)(P<0.05), and inhibited the activity of YAP/TEAD luciferase.At the same time, NVP-TNKS656 up-regulated two major negative regulators of YAP, namely AMOTL1 and AMOTL2 proteins, and promoted the apoptosis of HLE and HLF cells in a dose-dependent manner.
ConclusionNVP-TNKS656 can inhibit the proliferation of HCC by stabilizing AMOTL1/ AMOTL2 and down-regulating the YAP target gene.This study indicates that NVP-TNKS656 can be used as a potential drug for treating HCC.
Key words:
Hepatocellular carcinoma; NVP-TNKS656; Signal pathway
Contributor Information
Qiao Yu
Department of Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
Hu Zhehui
Party Committee Office, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
Yu Gang
Xi Cheng the Second Retirement Centre for Retired Cadres, Beijing 100034, China
Duan Beibei
Department of Pharmacy, the Sixth Medical Center, PLA General Hospital, Beijing 100048, China
Zhang Shuai
Department of Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
Zhao Yunbo
Department of Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
Zhang Zijin
Department of Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
Li Lin
Department of Oncology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
