Study Of Phloretin’s Effect On Treatment Of Ulcerative Golitis Associated Colorectal Cancer In An Inducible Mouse Model

ObjectiveColon cancer is one of the common malignant tumor in digestive system.It is the main reason for the killing of inflammatory bowel disease.The occurrence of colitis associated cancer has experienced a pathological process about’inflammation-dysplasia-cancer’.Therefore,to control the colon inflammation development is favorable for colon cancer prevention.Phloretin is the effective active ingredients in leaf of Strigose Hydrangea,has anti-inflammatory,anti-tumor,pharmacological activity.In order to further explore the pathogenesis of colitis associated cancer,we used DSS model and AOM/DSS model as the breakthrough point,studied the phloretin’s influence on STAT3 signaling pathways,to find out low and effective antitumor drugs to provide experimental basis.Methods and ResultsThis study will from two aspects,cell experiments and animal experiments,to explore the prevention of colitis associated cancer in phloretin.1.The cells experimentChoose six kinds of natural compounds which isolated from medicinal plants for active screening test on caco-2 cells in vitro.this study selected phloretin as the object for the further study by MTT method.Using flow cytometry detection technology to determine phloretin monomer influence on caco-2 cells’ proliferation,cycle and apoptosis,the results showed that phloretin is likely to be through to block caco-2 cells in vitro on S phase of cell cycle to induce caco-2 cells apoptosis and inhibit the further proliferation of colon cancer cells.Phloretin extraction concentrations treated on protein of caco-2 cells,western blot methods measuring the expression of IFN-y,IL-6,p-STAT3 and Bcl-xL,Bax protein,The results found that phloretin can decrease the STAT3 signaling pathway by influenced the IL-6,p-STAT3,Bcl-xL,IFN-γ protein’ s expression,and promote the expression of Bax protein.This indicates that the antitumor function of the phloretin may be associated with inhibition of STAT3 signaling pathway activation,and random quantity increase,the more obvious inhibition of STAT3 signaling pathway.2.The animal experimentsThrough the establishment of the acute ulcerative colitis model in Balb/c mice induced by DSS,to observe the influence in mice normally,DAI score,and the colonic mucosa tissue damage when used phloretin,then enzyme-linked immunosorbent(ELISA)to detect the serum TNF-α,IL-10 levels,and Western Blot method to detect the IFN-γ,IL-6,p-STAT3 and Bcl-xL,Bax protein’ s expression in colon tissue.The experimental results showed that phloretin can reduce the degree of inflammation in mice by adjusted the TNF-a,IL-10 levels,and inhibit tumor proliferation.Then continue to choose Balb/c mice as experimental animals object,established colitis associated cancer model in mice induced by AOM/DSS,observed the influence by used phloretin on mice model to its general behavior pattern,organization of general morphological observation,intestinal quality index and HE histopathological changes,then ELISA methods to detect the serum TNF-α,IL-10 levels and Western Blot method to detect the IFN-γ,IL-6,p-STAT3 and Bcl-xL,Bax protein’ s expression in colon tissue.According to the results,the size and number of colonic neoplasm of phloretin dosage group are significantly less than the model group,HE dyeing observation found that adenoma of intestinal mucosa decreased,inflammation have varying degrees of ease.Using the phloretin to intervent the AOM/DSS induced mice,we can be found that phloretin can through adjusted IL-6,p-STAT3,Bcl-xl,IFN-γ protein’ s expression,and raised its Bax protein’s expression in the colon-tumor tissue to suppress the activation of STAT3 signal pathways.According to the experiment,the data shows that the prevention effect of phloretin for colitis associated colorectal cancer is in a present positive correlation to concentration.ConclusionAccording to the experiment in vitro and vivo,we found that phloretin has obvious inhibitory effect on the caco-2 cells,inflammation and colon tumor tissue,and phloretin can improve the quality of the survival of mice through to prevent the further hyperplasia cancer of colon inflammation.Phloretin also can decrease IL-6,p-STAT3,Bcl-xl,IFN-y protein’ s expression,and increase Bax protein’ s expression level in caco-2 cells or colon tissue,which is one of the mechanism of inhibition of colon cancer.Phloretin can inhibit the activation of STAT3 signaling pathways,resulting in increased for continued the repair effects of chronic inflaammation of colon injury,inhibit inflammation-cancer dysplasia,quickens the process of the apoptosis of tumor cells,and then play its corresponding antitumor effects.