Mechanism Study Of Retro-zuojin Pill On Anti-tumor And Anti-metastasis

Malignant tumor, one of the most serious diseases threatening human life, has occupied the first place in the common cause of death in our country. At present, chemotherapy is the main method for cancer therapy, but it has indigenous defects, for instance,it has great toxicity, and multi-drug resistance is frequently occurred, etc. so, more and more people begin to look back on the traditional chinese medicine therapy, zuojin wan arise from “ ‘Dan Brook Heart Law’”,and be composed by Berberine and Evodia by 6:1,it has the effect of Qinggan xiehuo and descending perverse rise of stomach-qi to stop vomiting.Modern pharmacologocal studies have shown that: zuojin wan has a preferable role in prevention and treatment to gastric cancer, but it is still lacking theoretical foundation.In the former work of our laboratory,we had discouered and identified fanzuojin wan(be composed by Berberine and Evodia by 1:6) has more advantages in anti-tumor. In this study, we evaluated the effect of zuojin wan,fanzuojin wan and fufang(based on fanzuojin wan adding phytolaca americana and cinnamomum cassia Presl) on anti-tumor from overall,cellular and molecular levels,in order to provide theoretical support for fanzuojin wan used for prevention and treatment of cancer.In this study, MTT assay was used in vitro to detect the influence of different concentrations of ethanol extract from zuojin wan,fanzuojin wan and jiawei fufang on cell proliferation in human lung cancer cells(A549),cell scratch was used to determine the influence of these three compounds on cell migration rate, AO staining was used to test the cellular autophagy in starvation. In vivo experiments, we used H22 subcutaneous allograft model, Lewis experimental lung metastasis model and urethane-induced lung carcinogenesis model in mice to evaluate the effect of retro-zuojin Pill and fufang full-drug compatibility and water decoction on tumor prevention and treatment. By detecting the mental state, body temperature, survival rate, independent active number of times to assess the effect of tested drugs on TCM symptoms, we have observed the pathological changes of lung tissue by hematoxylin-eosin staining;and to investigate the changes of serum APT、AFu、Gp-73、SOD、MDA and LDH contents in tumor-bearing mice.In addition, western blot was used to observe the effect of tested drugs on expression of cancer stem cell marker Nanog and Oct4.In vitro, the experimental results showed that ethanol extracts from zuojin wan,fanzuojin wan and jiawei fufang can inhibit A549 cells proliferation in a dose dependent matter(IC50=47.3,39.1,32.6μg/ml),they can also inhibit cell migration,however,fanzuojin wan and jiawei fufang are more effective at the same dose.Besides, fanzuojin wan and jiawei fufang can also inhibit A549 cells autophagy in starvation.In vivo, the experimental results showed that fanzuojin Pill and jiawei fufang can slow tumor growth, prevent tumor metastasis and prevent tumorigenesis, maintain constant body temperatures and improve the quality of life in mice, reduce serum APT、AFu、Gp-73、MDA and LDH contents,but improve serum SOD content.,prolong the survival time of tumor-bearing mice.Pathological results showed that the Fanzuojin wan aqueous extract can significantly improve the pathologic changes of lung tissue.Western blot results showed that fanzuojin wan and fufang can significantly reduce the expression of tumor stem cell markers Nanog and Oct4 in lung of Urethane-induced lung carcinogenesis model mice.Above all, fanzuojin wan and jiawei fufang have a preferable role in stop tumor cell growth and prevent lung cancer in mice,it implies that hanning qiyu may play an important role in the occur and development progress of malignant tumours,and we should reduce the use of “cold” medicine in clinic tumor therapy.