Mechanism Study Of Alcohol Extract From Semen Pharbitidis On Anti-tumor And Anti-metastasis In Lewis Lung Cancer

Lung cancer is one of the pernicious tumours heavily harming human life and health. and is the cancer which causes the highest morbidity and mortality in the world. In recent years, the studies of tumor metastasis at home and abroad have made great progress, but there is no medicine which can be widely used in clinical prevention and treatment of tumor metastasis. It is very important to explore the therapeutical ideas of Chinese medicine about tumor metastasis and to clarify their possible mechanism. The Chinese medicine Semen Pharbitidis can clear phlegm, kill the parasites and eliminate the lumps in human bodies, and bring the stool out. It is possible for Semen Pharbitidis to have the anti-tumor and anti-metastasis effects in many aspects based on phlegm theorety from Chinese medicine. The purpose of this study are to fully work out the unique anti-tumor advantages of the Chinese medicine Semen Pharbitidis by its anti-tumor mechanism in system, cellular and molecular levels, and to provide the new experimental idea for the Chinese medicine in tumor treatment and prevention.In this study, MTT assay is used to determine the effect of alcohol extract from Semen Pharbitidis on cell proliferation in Lewis lung cancer cells, cell scratch is used to determine the cell migration rate, acridine orange staining is used to test the cellular autophagy. based on above observations, the anti-proliferation and anti-metastasis of Alcohol extract from Semen Pharbitidis are evaluated in vitro. In vivo, the subcutaneously transplanted model of Lewis lung carcinoma is established to evaluate the anti-proliferation and anti-metastasis of Alcohol extract from Semen Pharbitidis. The serums carcinoembryonic antigen (CEA) and p2-microglobulin (J32-MG) are detected by electrochemiluminescence. the influence of Alcohol extract from Semen Pharbitidis on the immune activity is investigated by the spleen index and thymus index.the tumor metastasis is determined by the number of metastatic nodules on the lung surface, the survival quality of mouse is evaluated by the automatic activity and the life span..Furtherly. Fluorescent yellow transmission is applied to test the intercellular junction communication, western bloting is used to test the cells expression of aquaporin1(AQP1), and immunohistochemistry is applied to detect the expression of AQP1and gap junction protein Cx43in lung metastatic nodles.In vitro, the experimental results showed that:Alcohol extract from Semen Pharbitidis inhibited cell proliferation in a dose-dependent manner with a half inhibitory concentration (IC50) of37.21[.ig■mL-1in lewis lung cancer ceils. Alcohol extract from Semen Pharbitidis could reduce cell autophagy under low serum condition and prevent the cells migration. The drug-treated lewis cells were migrated20.5■3.1um and46.8■4.2um respectively, while the control cells were migrated121.3■8.6umIn vivo, the experimental results showed that:Alcohol extract from Semen Pharbitidis could reduce the levels of the serum CEA and P2-MG in tumor-bearing mice, and obviously inhibited tumor growth with the inhibition rates of43.5%and38.2%in high dose group and in low dose respectively. In addition, Alcohol extract from Semen Pharbitidis could significantly prevent the lung metastasis in dose-dependent manner in the experimental pulmonary metaststic model. Compared with model group, the mouse treated with Alcohol extract from Semen Pharbitidis showed a prolonged survival and more automatic activity, but their spleen index and thymus index had not been affected, which suggested a higher survival quality in Semen Pharbitidis group.Mechanism study results showed:Alcohol extract from Semen Pharbitidis could reduce the cell protein AQP1. and promote the intercellular junction communication. In immunohistochemistry. Alcohol extract from Semen Pharbitidis could weaken the positive staining density of aquaporin AQP1and enhance the staining density of gap junction protein of Cx43in tumor-bearing lung tissue.In summary, Alcohol extract from Semen Pharbitidis has the exact role in anti-tumor and anti-metastasis, it is mechanism may be related to the enhancement of intercellular junction communication and the reduction of aquaporin AQP1.