Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 21, 2016; 22(15): 3962-3968
Published online Apr 21, 2016. doi: 10.3748/wjg.v22.i15.3962
Norcantharidin combined with ABT-737 for hepatocellular carcinoma: Therapeutic effects and molecular mechanisms
Jing Ren, Gang Li, Wen Zhao, Ling Lin, Tao Ye
Jing Ren, Gang Li, Ling Lin, Tianjin Key Laboratory of Biomedical Detection and Instruments, Tianjin University, Tianjin 300072, China
Jing Ren, Wen Zhao, Medical Laboratory, Tianjin Medical University General Hospital, Tianjin 300052, China
Tao Ye, Department of Oncology, Minhang Hospital, Fudan University, Shanghai 201199, China
Author contributions: Ren J and Ye T designed research; Ren J, Li G, Zhao W and Ye T performed research; Li G, Zhao W and Lin L contributed new reagents/analytic tools; Ren J, Lin L and Ye T analyzed data; and Ren J and Ye T wrote the paper.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Tianjin University.
Conflict-of-interest statement: We declare that there is no conflict of interest to disclose in our study.
Data sharing statement: We state that no additional data are available to share.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Tao Ye, MD, Department of Oncology, Minhang Hospital, Fudan University, No. 170 Xinsong Road, Minhang District, Shanghai 201199, China. yaplter@vip.sina.com
Telephone: +86-21-64923400 Fax: +86-21-64923400
Received: November 29, 2015
Peer-review started: November 30, 2015
First decision: December 16, 2015
Revised: December 30, 2015
Accepted: January 17, 2016
Article in press: January 18, 2016
Published online: April 21, 2016
Abstract

AIM: To study the therapeutic effect of norcantharidin (NCTD) combined with ABT-737 on hepatocellular carcinoma cells and the molecular mechanism.

METHODS: Two hepatocellular carcinoma (HCC) cell lines, HepG2 and SMMC-7721, were selected. ABT-737 and NCTD were allocated into groups to be used alone or in combination. HepG2 and SMMC-7721 cells were cultured in vitro. Liver cancer cells in the logarithmic phase of growth were vaccinated and cultured to the cell wall stage; these cells were treated for 48 h with different concentrations of NCTD, or ABT-737, or NCTD combined with ABT-737. The cell proliferation inhibition rate was detected by methyl thiazolyl tetrazolium. The expression of Mcl in HCC cells was detected by Western Blotting, and the cells in each group after treatment had apoptosis detected by flow cytometry. The proliferation inhibition rate, the expression of Mcl-1 in cells and the apoptosis inducing effect of treatment were observed in each group, and the effect of NCTD on ABT-737 in the treatment of HCC and its mechanism of action were analyzed.

RESULTS: As the concentration of NCTD increased, the cell proliferation inhibition rate gradually decreased; and the treatment effect of ABT-737 1-3 μm combined with NCTD on cell proliferation inhibition was stronger than that of ABT-737 alone. The difference was statistically significant (P < 0.05). In observing the expression of Mcl-1 in cells after the treatment of different concentrations of NCTD, this was partially inhibited after treatment with NCTD 15 μm, and the expression of Mcl-1 was almost undetectable after treatment with NCTD 30 μm and 60 μm. The effect on inducing apoptosis with the treatment of ABT-737 or NCTD alone for 48 h was lower than that of the control group. The difference was not statistically significant (P > 0.05). The effect on inducing apoptosis in HepG2 and SMMC-7721 cells with the treatment of ABT-737 combined with NCTD for 48 h was greater than that of ABT-737 or NCTD alone. The difference was statistically significant (P < 0.05).

CONCLUSION: NCTD combined with ABT-737 has a positive role in the treatment of HCC, and it has great value in clinical research.

Keywords: Norcantharidin, Hepatocellular carcinoma cell, Mcl-1

Core tip: The effects of ABT-737 and norcantharidin (NCTD) alone or in combination on HepG2 and SMMC-7721 cells were tested by methyl thiazolyl tetrazolium, Western blot and flow cytometry. We found that as the concentration of NCTD increased, the cell proliferation inhibition rate gradually decreased; and the treatment effect of ABT-737 1-3 μm combined with NCTD on cell proliferation inhibition was stronger than that of ABT-737 alone (P < 0.05). The effect on inducing apoptosis in HepG2 and SMMC-7721 cells with the treatment of ABT-737 combined with NCTD for 48 h was greater than that of ABT-737 or NCTD alone (P < 0.05). NCTD combined with ABT-737 has a positive role in the treatment of HCC.