Molecular targets of Chinese herbs: a clinical study of hepatoma based on network pharmacology

Abstract

Traditional Chinese medicine (TCM) has been used to treat tumors for years and has been demonstrated to be effective. However, the underlying molecular mechanisms of herbs remain unclear. This study aims to ascertain molecular targets of herbs prolonging survival time of patients with advanced hepatocellular carcinoma (HCC) based on network pharmacology, and to establish a research method for accurate treatment of TCM. The survival benefit of TCM treatment with Chinese herbal medicine (CHM) was proved by Kaplan-Meier method and Cox regression analysis among 288 patients. The correlation between herbs and survival time was performed by bivariate correlation analysis. Network pharmacology method was utilized to construct the active ingredient-target networks of herbs that were responsible for the beneficial effects against HCC. Cox regression analysis showed CHM was an independent favorable prognostic factor. The median survival time was 13 months and the 5-year overall survival rates were 2.61% in the TCM group, while there were 6 months, 0 in the non-TCM group. Correlation analysis demonstrated that 8 herbs closely associated with prognosis. Network pharmacology analysis revealed that the 8 herbs regulated multiple HCC relative genes, among which the genes affected proliferation (KRAS, AKT2, MAPK), metastasis (SRC, MMP), angiogenesis (PTGS2) and apoptosis (CASP3) etc.

Figure 1. Process overview.

Figure 2. Survival analysis between the TCM and non-TCM group.

The median OS in the TCM group was longer than that in the on-TCM group (13 vs 6 months, respectively; P < 0.001). OS = overall survival, TCM = traditional Chinese medicine.

Figure 3

The ingredient-target networks of (A) RST, (B) FC, (C) CT, (D) RS, (E) RAB, (F) RB, (G) SL and (H) RAM. The diamond nodes represent ingredients, and the circular nodes represent targets. The colors of the nodes are illustrated from red to yellow in descending order of degree values.

Figure 4. The herb-target networks of the 8 herbs.

The diamond nodes represent herbs, and the circular nodes represent targets. The targets distributed in a circle represent they are acting by the same number of herbs, which illustrated as “n”.

Figure 5. Simplified pathways in HCC.

All of the targets are shown by the gene name.

Figure 6. Effect of the aqueous extract of FC on cell proliferation and cell migration.

Data are expressed as mean ± S.D. (A) The morphology of SMMC-7721 cells (upper) and the result of wound-scratch assay (lower). (B) The results of Typanblue staining assay (left) and CCK-8 assay (right), “***” represented for p < 0.001. (C) Immunocytochemical staining showed Erk, p-Erk, Akt and p-Akt of four groups. (D) Western blot assay analyzed Erk, p-Erk, Akt and p-Akt of four groups.