Ginsenoside-Rh2 Potentiates The Sensitivity Of Tumor Cells To Betulinic Acid

At present, cancer is a serious threat to human and animal health, which is a serious threat to the health of people. Fragnent, the main treatment for cancer is surgery, supplemented by radiotherapy, chemotherapy and other methods. The efficacy of chemotherapy is proportional to the dose within a certain range. As the increases the risk of side effects occurred a corresponding increase in the tumor cells to chemotherapeutic drugs, tolerance, often lead to treatment failure. Found a synergistic effect of anticancer drugs can make drugs safe in therapeutic doses to generate better results. Most of the anti-tumor treatment, mainly including chemotherapy through induced apoptosis, and to prevent the body from drug tolerance, two and multi-drug combination therapy in the treatment of malignant tumors is better than a single anti-cancer drug. Because of synergies between the various drugs help to enlarge the original weak death signal. Therefore, the drug combination treatment may be a strong therapeutic strategy.Ginsenoside Rh2 (ginsenoside Rh2, G-Rh2) is extracted from ginseng with natural active, the study confirmed that the composition has a strong anti-tumor activity. According to reports, G-Rh2 can induce human breast cancer cells, human melanoma cells, human prostate cancer cells, human leukemia cells, human liver cancer cells and other tumor cells. Ginsenoside Rh2 also induces tumor cell differentiation, enhance immunity, and improve the ability of anti-tumor effect.Betulinic acid [3β-Hydroxy-lup-20(29)-en-28-oic acid] is a lup-type pentacyclic triterpenoids. Betulin acid is the carboxylic acid derivatives of C-28 of betulinol. However, betulinic acid with more people expected pharmacological activity which diference with betulin, such as anti-inflammatory, anti-cancer, anti-HIV and so on. Betulinic acid has anti-tumor, anti-HIV, anti-inflammatory effects, anti-malarial in vitro and other diseases.In this study, we found ginsenoside Rh2 significantly increased sensitivity to betulinic acid in A549 cells and HepG2 cells. Through analysis of cell morphology of two cell lines joining the two drugs, the morphological change significantly, there bubble membrane, nuclear condensation phenomena such as apoptosis. DAPI nuclear staining can be more easily observed phenomenon. Based on the above phenomenon, we analyzed by flow cytometry showed that, cells in the Sub-G1 significantly increased the percentage of the number when G-Rh2 and Bet A join to A549 cells and HepG2 cells simultaneously,. Caspase-3 activity analysis showed that, cells joined G-Rh2 and Bet A at the same time increased the activity of caspase-3, compared with G-Rh2 and Bet A were added alone, and the activity of caspase-3 increased significantly as the time. Using Western-Blot analysis PARP that is substrate of caspase-3 , the results show that the fracture of PARP when G-Rh2 and Bet A added at the same time increased significantly. Using Western-Blot analysis of caspase-9 activation, the results showed that activation of caspase-9 that G-Rh2 and Bet A all in tumor cells is higher than G-Rh2 and Bet A alone in A549 cells and HepG2 cells, and with time, activation is more evident.In summary, we found that ginsenoside Rh2 can improve the A549 cells and HepG2 cells to betulinic acid sensitivity, and confirmed that the result is caused by tumor cells through programmed cell death occurred.