Study Of Molecular Mechanism On Compound Prescription Of WeiMaiNing Inhibiting Lung Cancer Metastasis

The process of tumor metastasis consists of many factors and many steps. Adhesionbetween tumor cells and endothelial cells and tumor angiogenesis are the critical steps of tumorblood metastasis. According to the theory of traditional Chinese medicine, clinical practices andpast investigative results, we chose WeiMaiNing,berberine and HuangQi polysaccharide, whichhas the anti-lung cancer, anti-various tumors and immune regulation function, respectively, andconstituted the Chinese medicine compound prescription(FWN). The effect of FWN onanti-proliferation, anti-metastasis in lung cancer and molecular mechanism were investigated invivo and vitro, so as to provide the academic bases for anti-metastasis drugs researching,development and applying in clinic. This study consists of three parts: 1) The effect of each constitute medicine of FWN on inhibiting tumor cells in vitro, whichprovides the experimental bases for constituting prescription of FWN. 2) Anti-lung cancer experiment in vivo: ① The effect of FWN on inhibiting murine Lewislung carcinoma in vivo: To establish murine Lewis lung carcinoma (3LL) transplant model,observe changes of the body weight and inhibitory rate after FWN treatment, and detect theeffect of FWN on cell cycle.② The effect and molecular mechanism of FWN on suppressingspontaneous lung metastasis in 3LL in vivo: To observe the effect of FWN on spontaneous lungmetastasis focus formation, and detect the effect of FWN on adhesion molecule expression intumor cell and on tumor angiogenesis. 3) Anti-lung cancer experiment in vitro:① The effect of FWN on anti-proliferation inPG,PAa: To investigate the effect on cell cycle. ② The effect and molecular mechanism ofFWN on anti-tumor metastasis in vitro: To observe the effect of FWN on adhesion between highmetastasis lung cancer cell(PG) and endothelial cells, detect the effect of FWN on adhesionmolecule expression in PG, HUVEC and on tumor angiogenesis. The animal experiment was divided randomly into six group: nomal saline solution(NS),Berberine(Ber),WeiMaiNing lower dose(WML),WeiMaiNing higher dose(WMH),compoundprescription of WeiMaiNing(FWN) and Cyclophosphamide(CTX). The cultured PG cell was divided into five group: control, Berberine(Ber),WeiMaiNing(WMN), compound prescription of WeiMaiNing lower dose(FWL) and compoundprescription of WeiMaiNing higher dose(FWH).The cultured HUVEC cell was divided into six英文摘要 5group: It included TNF-α(or bFGF) group besides above five group in PG.The results displayed as follow:1 The effect of each constitute medicine of FWN on inhibiting tumor cells in vitro Berberine(20,40μg/ml)and WeiMaiNing(125,250μg/ml)displayed inhibitory effecton six tumor cells chose at different degree by MTT, but HuangQi polysaccharide had not effecton them.2 Anti-lung cancer experiment in vivo2.1 The effect and molecular mechanism of FWN on inhibiting murine Lewis lung carcinomain vivo Inhibitory rate of WMN taken orally in 3LL was from 33.59% to 40.63% at a dosageranging 120~200mg/kg?d. Inhibitory rate of berberine taken orally was 15.09% at 20g/kg d.Inhibitory rate of FWN taken orally was increased greatly up to 47.66%.The result indicates thatthis three drugs cooperate with each other in inhibitory effect. FWN could block 3LL cells inG0-G1 phase by Flow cytometry(P<0.01). FWN could significantly decrease Cyclin D1expression in 3LL cells by immunohistochemistry staining and Western blotting(P<0.01).2.2 The effect and molecular mechanism of FWN on suppressing spontaneous lung metastasisin 3LL in vivo2.2.1 The effect of FWN on spontaneous lung metastasis focus formation In FWN group, lung metastasis focus was smaller and fewer, formation rate of spontaneouslung metastasis focus was 60%(P<0.01).This indicates that FWN has obviously inhibitoryeffect on lung metastasis in 3LL.2.2.2 The molecular mechanism of FWN on suppressing spontaneous lung metastasis in 3LL2.2.2.1 The...