Analysis of the mechanism of action and in vitro experimental verification of resveratrol against multiple tumors based on network pharmacology
Ruixiangsu is extracted from plants of the Ruixiang genus and is a natural medicinal monomer first developed in China. Due to its anticoagulant properties, it is currently mainly used in clinical practice for the treatment of cardiovascular diseases. In addition, a large number of studies have shown that daphnetin has inhibitory effects on a variety of tumor cells. Its mechanisms include inhibition of breast cancer through MAPK1/2-JNK1/2-Akt pathway, inhibition of liver cancer through Bcl-2/Bax target, inhibition of lung adenocarcinoma through Akt/NF kB pathway, and inhibition of ovarian cancer through AMPK/Akt/mTOR pathway. The targets and pathways of daphnetin’s anti-tumor effect have some commonness. Tumor is a major disease that seriously endangers human health, and its incidence rate is increasing year by year. Among them, liver cancer is the most lethal malignant tumor; Triple negative breast cancer is a kind of female breast cancer with high malignancy and difficult treatment; Malignant glioma is a neurological malignancy with high rates of disability and mortality. Previous studies have shown that resveratrol has clear anti-tumor effects, but whether there are common targets or pathways for its anti-tumor effects has not been reported. Therefore, this study intends to predict the common target or pathway of daphnetin in the treatment of malignant glioma, liver cancer and triple negative breast cancer through network pharmacology, and verify it in vitro, so as to provide experimental basis for the clinical application of daphnetin in anti-tumor.

In this study, we first used network pharmacology analysis to find that P53 pathway is the common pathway of daphnetin against malignant glioma, liver cancer and breast cancer. Meanwhile, this study found through in vitro validation experiments that resveratrol significantly inhibited the proliferation of U-251MG, HepG-2, and MDA-MB231 cells at concentrations of 40 and 80 μ g/mL, and significantly increased the expression level of P53 protein in these three tumor cells. P53 is an important tumor suppressor gene, whose mechanisms mainly include cell cycle arrest, promoting tumor cell aging and apoptosis, inhibiting angiogenesis and tumor metastasis, and promoting DNA synthesis, repair, and regulation. Our research results indicate that resveratrol can increase the expression of P53 in the three types of tumor cells mentioned above, thereby exerting anticancer effects. RRM2 is a key enzyme regulating DNA synthesis and repair in P53 pathway, and it is enriched in P53 pathway. Numerous studies have also shown that P53 can regulate cell cycle, DNA synthesis and repair, and induce tumor cell apoptosis through RRM2. Therefore, RRM2 has become an important target for various cancer treatments.

As a result, it was found that the expression of RRM2 protein was significantly reduced in U-251MG, HepG-2, and MDA-MB231 cells treated with 40 and 80 μ g/mL resveratrol. RRM2 is the most important subunit of ribonucleic acid reductase (RR) and a key enzyme that catalyzes DNA synthesis and repair in living organisms. Current research shows that RRM2 is highly expressed in glioma, breast cancer, liver cancer and other diseases, and is the target of tumor treatment. It is related to the invasion and metastasis ability of tumor cells, drug resistance and cell cycle regulation. RRM2 contains tyrosine free radicals and Fe-S groups, so most of the targeted drugs currently developed are based on these targets, such as free radical scavengers, iron chelators, iron analogues, etc. Some RRM2 inhibitors have been applied in the clinical treatment of tumors, but they have various side effects such as blood lymphatic system metabolic disorders, liver and kidney dysfunction, gastrointestinal reactions, etc. Ruixiangsu is a natural plant active ingredient with iron chelation function. Previous studies have found that Ruixiangsu can inhibit RR activity and expression in malaria parasites. Our results further demonstrate that Ruixiangsu can inhibit the expression of RRM2 protein in various human tumor cells. Due to the important role of RRM2 in DNA replication and repair, it is speculated that resveratrol can inhibit various tumor cells and is closely related to its regulation of RRM2 expression. Ruixiangsu can be used as a new RRM2 inhibitor for the treatment of tumors, and it has the characteristics of high safety and wide distribution.
In summary, our results indicate that resveratrol can inhibit the proliferation of U-251MG, HepG-2, and MDA-MB231 cell lines, and has good binding ability with P53 and RRM2. The mechanism by which resveratrol inhibits multiple tumors may be related to the P53/RRM2 pathway. Ruixiangsu can be used as a new RRM2 inhibitor with great development prospects.