The Biological Efficacy Of Cafestol’s Inhibition Of Colon Cancer

Objectives:The research starts from the in vitro cell level to explore influence of cafestol on proliferation of colon cancer cell HCT116 and its dose-effect relationship,detect autophagy marker to confirm that cafestol induces colon cancer cells to lead to autophagic cell death,so as to delve into the key pathway and action mechanism of cafestol inducing autophagy via detecting key target gene on different autophagical pathways.Exploration into the biological efficacy of cafestol inhibiting colon cancer conduces to further develop cafestol resources,and display the function of cafestol in preventing and controlling colon cancer.This is of far reaching importance to advance medication and enhance the key function of fundamental research to prevention and control of chronic diseases.Methods:Cultivated cells HCT116,added cafestol of different dosages(Oμm,10μm,20μm,40μm,80μm and 160μm)to be incubated with cells for 48h,detected influence of cafestol on proliferation of colon cancer cells HCT116 and its dose-effect relationship via MTT experiment,detected autophagical marker using transmission electron microscope,then used Western Blot to detect change of expression of LC3B-Ⅱ induced by cafestol,lastly detected key target gene on different autophagical pathway using RT-PCR.Results:1.MTT result shows that with escalation of cafestol concentration,the cells activity was inhibited,i.e.cafestol can inhibit proliferation of colon cancer cells.2.The result of transmission electron microscope shows that after treating HCT116 cells using cafestol of different dosages(0μM,20μM,40μM)for 24h,the mitochondria deformation for cells incubated with 40μm cafestol was observed under transmission electron microscope,and they have autophagosome encircled by bilaminar membrane,indicating cell autophagy occurred to HCT116 cells under inducement of cafestol.2.Western Blot detected autophagical marker LC3B-Ⅱ.The result shows that with escalation of cafestol concentration,strap trace of LC3B-Ⅰ faded gradually.While LC3B-Ⅱ is enhanced gradually.So it was conjectured that cell autophagy occurs to colon cancer cells when cafestol achieves a certain concentration.4.PCR result show that with gradual rise of cafestol concentration,the relative change of expression level of Beclinl and mTOR has no statistical differences.It was conjectured that autophagy of colon cancer cell HCT116 may be not via these two pathways.Concrete key target spot need further study.Conclusions:1.Cafestol can inhibit proliferation of colon cancer cells.2.Cafestol of certain concentration can induce autophagy of HCT116 cells.3.The relative change of expression level of Beclinl and mTOR in different concentrations of cafestol has no statistical differences,prompting that the cell autophagy induced by cafestol may be not induced by the two genes.The concrete key target spot needs further studies.