Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (19): 3016-3023.doi: 10.3969/j.issn.2095-4344.3523

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Chinese medicine compound preparation inhibits proliferation of CD133+ liver cancer stem cells and the expression of stemness transcription factors

Qin Yanchun1, Rong Zhen2, Jiang Ruiyuan1, Fu Bin1, Hong Xiaohua1, Mo Chunmei2   

  1. 1Department of Graduate, Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China; 2First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China
  • Received:2020-05-11 Revised:2020-05-15 Accepted:2020-06-29 Online:2021-07-09 Published:2021-01-13
  • Contact: Mo Chunmei, Chief physician, First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China
  • About author:Qin Yanchun, Master candidate, Department of Graduate, Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81760850 (to MCM); the Basic Ability Improvement Project of Young and Middle-Aged Teachers in Guangxi Universities, No. 2018KY0282 (to MCM); the Guangxi Medical and Health Appropriate Technology Development and Application Project, No. S2018046 (to MCM); the Research and Development of Hospital Preparations in the First Affiliated Hospital of Guangxi University of Chinese Medicine, No. 2018ZJ004 (to MCM)

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Abstract: BACKGROUND: The malignant transformation of cancer originating from stem cells is increasingly recognized. Liver cancer stem cells have the potential of continuous self-renewal and multi-directional differentiation, which can promote the occurrence and development of liver cancer, postoperative recurrence and the production of drug resistance.
OBJECTIVE: To explore the effect of Fuhe preparation formula on proliferation of CD133+ liver cancer stem cells and stemness transcription factor expression. 
METHODS:  CD133+ HepG2 liver cancer stem cells were selected by immunomagnetic bead method. CD133+ HepG2 cells were treated with serum containing 2%, 4%, 8%, 10%, 12% and 16% Fuhe preparation formula respectively. The inhibitory rate of CD133+ HepG2 cells was determined by CCK-8 method, and the optimal volume fraction of serum containing drugs was selected. The CD133+ HepG2 liver cancer stem cells were divided into dimethyl sulfoxide group, application and preparation group, serum control group and blank control group. Cells in the dimethyl sulfoxide group were intervened with 0.05% dimethyl sulfoxide. Cells in the application and preparation group were intervened with 16% application and preparation containing serum. Cells in the serum control group were intervened with 16% normal rat serum. After 6 days of intervention, the percentage of CD133+ HepG2 liver cancer stem cells was detected by flow cytometry. The mRNA and protein levels of transcription factors SOX2, NANOG, and OCT4 were detected by RT-PCR and western blot assay. 
RESULTS AND CONCLUSION: (1) The serum containing application and preparation formula inhibited the proliferation of CD133+ HepG2 cells, which was related to the time and concentration. The inhibitory effect of 16% application and preparation formula was the most obvious in 72 hours (P < 0.05). (2) Compared with the serum control and blank control groups, the percentage of CD133+ HepG2 liver cancer stem cells was decreased in the application and preparation and dimethyl sulfoxide groups (P < 0.05), and the effect was more obvious in the application and preparation group. (3) Compared with the serum control and blank control groups, SOX2, NANOG, OCT4 mRNA and protein expression levels were downregulated in the application and preparation and dimethyl sulfoxide groups (P < 0.05), and the effect was more obvious in the application and preparation group. (4) These findings conclude that the serum containing application and preparation formula can effectively reduce the expression levels of CD133, a marker on the surface of liver cancer stem cells, and transcription factors SOX2, NANOG and OCT4, and can reduce the malignant degree of liver cancer stem cells. 

Key words: stem cells, liver cancer stem cells, application and preparation formula, drug serum, stem cell transcription factors

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