Exploring The Anti-tumor Immune Responses Mechanisms Of Lingjia Anti-tumor Medicine In Tumor-bearing Mice

Chinese medicine is a treasure of the Chinese nation, is explore the long-term struggle with the disease out of the great achievements, in the reproduction and the development of the Chinese medicine play a very important role. In contemporary China, the traditional Chinese medicine and Western medicine has become the main means of disease treatment. In recent years, with the rapid development of Chinese traditional medicine industry, the use of Chinese traditional medicine and natural small molecules to tumor therapy which has been more and more favored in the medical profession. In the treatment of tumor, the Chinese traditional medicine has the unique advantages, for example, improve immune system in cancer treatment, reduce side effects of chemotherapy, improve the survival of patients after treatment. However, due to the complexity of Chinese medicine prescription, in the process of treating diseases treatment mechanism is not clear, so that traditional Chinese medicine promotion and large-scale clinical application.Regulatory T cells (Treg) is a special kind, of immune cells in T cells, which play a very important role in the process of tumor occurrence and development, and has its own immune, immune tolerance and immune suppression function, which can made the antitumor immune response by the synergy of various mechanisms. Therefore, during the treatment of malignant tumor, as a traditional unique form in traditional Chinese medicine, it can inhibit the Treg cell number and weaken the function of Treg cells, restrain the immune tolerance, improve immunity, which also provides a new idea for the treatment of Chinese traditional medicine on tumor.Lingjia Anticancer Medicine(LAM)is anticancer compound traditional Chinese medicine which has already used for many years in clinical, which improve symptoms, enhance quality of life and prolong the survival time, during the treatment of hepatocellular carcinoma and breast cancer patients, the effect is obvious. The previous experimental results show that, the ethanol extract (LAM) can inhibit the proliferation of tumor cells in vitro experiment, the activation of NF-κB signaling pathway and the expression of downstream gene, and the cycle block of tumor cell finally induce the apoptosis of tumor cells. The main purpose of this study is to study the ethanol extract (LAM) to enhance anti-tumor immunity by regulating the immune system. Firstly, the daily oral administration of Chinese traditional medicine got by important traditional decocting method, and then got LAM by 45% ethanol and 80% ethanol extract liquid. In order to evaluate the extraction fractions (LAM) of toxicity on mice, we were divided the mice into four groups, which group has three different concentrations (0.5 g/Kg,1 g/Kg,2 g/Kg) of LAM to feed mice. After 14 days, there is no obvious effect for mice normal growth, no mice died, which according to the toxicological evaluation standard, so LD50>2 g/Kg. The result of liver and kidney of mice HE staining showed that, three concentration LAM is not damaged, and changing the compound extract non-toxic for mice safe. To verify the control response of LAM on the immune function, we construct a BALB/c-4T1 mouse model of breast cancer. The experiment divided into six groups:normal group (Normal), bearing cancer control group (Control), prevention group (Pre-5 g/Kg), the treatment group one (5 g/Kg), the treatment group two (10 g/Kg), cyclophosphamide group (CPM-10 mg/Kg).We found that the prevention group, treatment group and CPM group were significantly inhibit the growth of tumor, and the Antitumor effect of prevention group (67.9%) was better than the same concentration of the treatment group (61.54%) and cyclophosphamide group (43.08%).At the same time, the prevention group, treatment group, the CPM group, the inhibitory effect of Treg cell is clear, by the number of Treg cell from Immunophenotype CD4+CD25+FOXP3+ and CD4+CD25+CD127.The result of ELI SA is that, LAM significantly inhibit the secretion of IL-10 and TGF-β and the expression of FOXP3 was obviously inhibited after treatment confirmed by Western blotting. The experiment deeper into verified that the drugs effect on cell proliferation, CD8 cells and NK cells which inhibited by Treg cells. The results showed that LAM can improve the proliferation of lymphocyte, the CD4/CD8 tended to be normal, the percentage of NK cells increased, the ELISA results also indicated that the secretion of IL-2 and IFN-y, perforin and granzyme B increased significantly.Given all that, On the basis of in vitro experiments, in vivo experiments confirmed, LAM can inhibit tumor growth by inhibiting Treg cell number and function, break caused by Treg cells of immune tolerance and improve the body immune function, restore the normal antitumor immunity. These discussing a possible anti-tumor mechanism of Lingjia Anticancer Medicine extracts, For Lingjia Anticancer Medicine clinical application provides the theory support.