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Bie Jia Jian Pill Combined with Bone Mesenchymal Stem Cells Regulates microRNA-140 to Suppress Hepatocellular Carcinoma Stem Cells
International Journal of Stem Cells
Published online February 28, 2021;  
© 2021 Korean Society for Stem Cell Research.

Huang Jingjing1,2,*, Huang Hongna3,*, Zhang Wenfu4, Lv Jianlin4, Huang Guochu1, Lin Yuanjia1, Chen Songlin5, Hu Yueqiang3,6

1Department of Spleen, Stomach and Liver Diseases, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China
2Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine, Nanning, China
3Teaching and Research Office of Internal Medicine of Traditional Chinese Medicine, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China
4Department of Internal Medicine, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China
5Department of Human Resources, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China
6Guangxi Key Laboratory of Basic Research of Traditional Chinese Medicine, Nanning, China
Correspondence to: Hu Yueqiang
Teaching and Research Office of Internal Medicine of Traditional Chinese Medicine, The First Affiliated Hospital of Guangxi University of Chinese Medicine, No. 89-9, Dongge Road, Qingxiu District, Nanning 530200, China
Tel: +86-13152510678, E-mail: huyq@gxtcmu.edu.cn
*These authors contributed equally to this work.
Received September 28, 2020; Revised December 10, 2020; Accepted January 17, 2021.
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background and Objectives: Cancer stem cells (CSCs) with tumorigenic potential are reported as the crucial factors of hepatocellular carcinoma (HCC) recurrence and therapy resistance. Bone mesenchymal stem cells (BMSCs) are documented to play an important role in the protection of hepatocytes. Bie Jia Jian pill (BJJP), a Traditional Chinese Medicine, has been used to treat liver fibrosis and liver cancer. This study aimed to explore the potential role of combined use of BJJP with BMSCs in HCC cell lines.
Methods and Results: Flow cytometry was used to identify BMSCs isolated from BALB/c mice and CSCs enriched from Huh7 cells by measuring CD24, CD133, CD44, CD73, CD105, CD166, CD29, CD14 and CD34. Differentiation potential of BMSCs was also determined. Cell viability and proliferation ability of CSCs were determined by CCK-8 assay and clone formation assay. The expressions of CSCs biomarkers and Wnt/β-catenin signal pathway related proteins were determined by PCR and western blot. TOP-Flash/FOP-Flash luciferase assay was applied to measure the activity of β-catenin/TCF. Compared with untreated CSCs, BJJP or BMSCs treatment alone on CSCs lead to increased miR-140 expression and cell apoptosis, as well as decreased expressions of CD24, CD133, EpCAM and cell viability. Downregualted expressions of Wnt/β-catenin signal pathway related proteins, Wnt3a and β-catenin were found in response to BJJP or BMSCs treatment alone. The combination of BJJP+BMSCs treatment on CSCs could further enhance the suppressive effect on CSCs. Down-regulation of miR-140 in CSCs partially blocked the effects of BMSCs or BMSCs+BJJP on the expressions of Wnt3a and β-catenin as well as the cell viability and apoptosis of CSCs. Reversed expression pattern was found in CSCs transfected with miR-140 overexpression.
Conclusions: Taken together, we demonstrate that BJJP+BMSCs together could further enhance the suppressive effect on CSCs through regulating miR-140 and suppressing Wnt/β-catenin signal pathway. This study demonstrated the potential of BJJP+BMSCs in therapeutic treatment of HCC.
Keywords : Hepatocellular carcinoma, Cancer stem cells, Bie Jia Jian pill, Bone mesenchymal stem cells, microRNA 140