Compatibility mechanism of Chrysanthemi Indici Flos compound against hepatocellular carcinoma based on balance of DNMTs/TET2

Abstract

Objective

At present, traditional Chinese medicine (TCM) compounds is widely used in tumor therapy. However, due to the current single and simplistic efficacy evaluation indicators that are detached from the guidance of TCM theory, the elucidation of its mechanisms of efficacy is hindered. Therefore, the compatibility mechanism of Chrysanthemi Indici Flos compound (YJHFF) against hepatocellular carcinoma based on DNA methyltransferases/DNA demethylase ten-eleven translocation 2 (DNMTs/TET2) balanced relationship was explored according to the concept of the overall balance of TCM.

Methods

This study screened the best compound compatibility ratio in the in vitro experiments, cell viability and DNMTs/TET2 balance were used as evaluation indicators to investigate the effects of single herb and different compatibility of compound on HepG2 and H22 cells. In vivo experiments were conduct on H22 liver orthotopic transplantation model mice. After treatment, the tumor tissues were observed by hematoxylin-eosin (HE), TUNEL and Ki67 immunohistochemical staining, respectively. The oxidative stress and liver function indicators in serum were detected by enzyme-linked immunosorbent assay. In addition, the proportion of tumor-infiltrating lymphocytes was detected by immunofluorescence staining. The mRNA expression levels of DNMTs and TET2 in tumor tissues were detected by RT-PCR.

Results

The results of in vitro experiments showed that the cell proliferation inhibition rate was highly consistent with the change trend of DNMTs/TET2 balance, accordingly, this study obtained the optimal formula YJHFF. In the in vivo experiments, the tumor growth inhibition rate of YJHFF in 5 g/kg was 62.97%. Extensive necrosis, increased apoptosis and inhibited proliferation activity were observed in tumor tissue. Furthermore, the serum SOD activity was significantly increased, MDA, ALT, AST and AFP levels were significantly decreased, suggesting YJHFF could systematically alleviated oxidative stress and improved liver function. Besides, the infiltration of CD3⁺, CD4⁺ and CD8⁺ T lymphocytes increased significantly, indicating that the body’s anti-tumor immune response was enhanced. Finally, RT-PCR analysis confirmed that antitumor effects were closely related to the core mechanism of DNMTs/TET2 expression regulation and reconstruction of epigenetic balance.

Conclusion

This work confirmed the rationality and feasibility of the DNMTs/TET2 balance relationship as an evaluation indicator for the study of the compatibility mechanism of TCM compounds. The YJHFF screened under the guidance of DNMTs/TET2 balance has liver protection and tumor immune function, showing good anti-hepatocellular carcinoma effect.