The Separation Of The Anticancer Bioactive Fractions Of AMH And Their Anti-cancer Effects

Objectives:Separating the anticancer bioactive fractions from the fungus plant extract AMH,,and to study the anticancer effects of the bioactive fractions both in vitro and in vivo.Methods:Extract the active components of fungalplant by ethanol circumfluence extraction and organic solvent. Cell culturing human lung cancer A-549cells, human breast cancer MCF-7and human colon cancer HCT-116, detect the inhibitory rate of cell proliferation, IC50, and draw it’s time-effect and dose-response curve. To evaluate the anti-cancer effect of active components and their combination drugs DDP,5-FU and clinical tumor from the cell level. Transplanted3LL micetumor model of human lung cancer cell line In C57BL/6J mice, randomly divided the mice into four groups:The control group and AMH-D treatment group (50,100,200mg/kg), administered by intraperitoneal injection every other day, were administered10times, measuring tumor weight, tumor inhibition rate was calculated and the spleen coefficient; selected Kunming mice, establishment of sarcoma S-180cell transplantation tumor model, the mice were divided into four groups randomly. The control group and AMH-D treatment group (50,100,150mg/kg), administered by intraperitoneal injection every other day, were administered5times, measuring tumor weight, tumor inhibition rate was calculated; establishment of sarcoma S-180cells of mice transplanted tumor model, mouse were randomly divided into control group, AMH-D (25,50,100mg/kg) treatment group, CP group (20mg/kg) and combination group, administered by intraperitoneal injection were administered every other day,5times, measuring tumor weight, tumor inhibition rate was calculated and the Q value, evaluation of fungal plant extract from the overall level (AMH-D) anti-cancer effect.Results:1) extraction, separation of AMH-D from fungi plants (2.03kg) and AMH-L (0.58kg).2) AMH-D (6.25,12.50,25and50μg/ml) on human lung cancer A-549cell proliferation inhibition rates were9.6%,29.29%,48.97%and86.38%, showed a significant dose-effect and time-effect relationship. AMH-D and DDP combination, the maximum Q value were1.14; and5-FU combination, the maximum Q value of1.17.3) AMH-D (50,100and200mg/kg) inhibited subcutaneous lung cancer3LL cell xenograft tumor growth inhibition rates were30.94%,34.11%and42.02%, each dose group of mice spleen coefficient compared with the control group, no significant difference (P>0.05). AMH-D (50,100and150mg/kg) on mice sarcoma S-180cells subcutaneous transplantation tumor inhibition rates were36.04%,43.14%and50.45%, presents the obvious dose-effect relationship.50mg/kg AMH-D and CP combination, Q value is1.27.4) AMH-L (2.50,5,10,15and20μg/ml) on A-549human lung cancer cells72h proliferation inhibition rates were94.88%,94.95%, and95.33%94.89%94.62%; on MCF-7human breast cancer cell72h proliferation inhibition rates were86.37%,87.18%,88.96%,92.20%and94.14%; on human colon cancer HCT-11672h cell proliferation inhibition rate was99.29%,99.83%,99.62%,99.65%and99.83%respectively. A-549, MCF-7and HCT-116three kinds of tumor cells of72h IC50were1.74μg/ml,2.08μg/ml and1.51μg/ml.Conclusion:1) AMH-D A-549on the proliferation of lung cancer cells significantly inhibited, and presents the obvious dose-effect relationship and time-effect relationship.2) AMH-D and DDP against human lung cancer A-549has additive effect, have synergistic effect with5-FU.3) AMH-D has inhibitory effect on lung cancer C57BL/6J3LL cells in mice and Kunming mice sarcoma S-180cell xenograft tumor growth, presents the obvious dose-effect relationship; AMH-D and CP combined with mouse sarcoma S-180cell transplantation tumor effect, synergistic action.4) AMH-L can significantly inhibit human lung cancer A-549, human breast cancer MCF-7and human colon cancer HCT-116cells proliferation.5) Inhibition of AMH-L on proliferation of lung cancer A-549cells by AMH-D.