The E-cadherin And PAR-3Expressions In The Process Of Colon Cancer Along With MicroRNA Mediation And Chinese Medicine Effects

Objective1.To explore the different expression and distribution of the characteristic molecular markers of Vimentin, CK18and CD133in the tissues of different distance to colorectal cancer;2.To detect the variation of E-cadherin and PAR-3expression and distribution differences in the tissues of different distance to colorectal cancer;3.Inducing the Organization structure change of neonatal rats colon with3D culture system, to detect the variation of E-cadherin and PAR-3expression and their related microRNA expression before and after treated with evodiamine and berberine,to explore whether evodiamine and berberine can reverse the decreased expression of E-cadherin and PAR-3.Methods1.To get tissue samples from different distances of proximal tumor focus, we called distance of10cm from tumor focus as number l,the distance of5cm as number2,then the distance of2cm as number3, Observing the variation of crypt core micro-archelloteture of three parts of tissue, using immunhistochemistry to detect expression and distribution of the Vimentin, CK18and CD133in the tissues of No.1,No.2and No.3.2.E-cadherin and PAR-3can be detected in immunhistochemistry and Western blot in the tissues of No.1,No.2and No.3.3.Inducing the Organization structure change of neonatal rats colon with3D culture system by TGF-β for48h, to detect the variation of E-cadherin and PAR-3expression and their related microRNA expression with qPCR. After that the model tissues was treated with evodiamine and berberine for6h and24h, then to detect the variation of E-cadherin and PAR-3expression and their related microRNA expression again with qPCR after treated with4.6mg/L evodiamine and42.8mg/L berberine for6h and24h.Results1.The expression and distribution of cell CD133from different parts of the crypt were detected by immunohistochemical method, The results showed that the expression of CD133existed in all the tissues, and the expression was mostly distributed in the crypt cell membrane and the cytoplasm. With the distance getting closer to tumor focus, The expression of CD133gradually increased,the datas between two adjacent groups had statistical significance.2.Vimentin expressed in all the tissues, and the expression mostly distributied in the cytoplasm of cells outside of the crypt. With the distance getting closer to tumor focus, The expression of Vimentin gradually increased,the datas between two adjacent groups had statistical significance.3.The expression of CK18was lowest in the No.2tissue, the highest was No.1tissue, the datas between two adjacent groups,group1and group3all had statistical significance.4.With the distance getting closer to tumor focus,crypt architecture was destroyed more and more obvious. It showed as the cells of crypt were irregularly arranged, some of them were lost.5.Western blot and immunhistochemistry both showed that the expression of E-cadherin and PAR-3decreased progressively from far to near to tumor focus.Immunhistochemistry showed that the trafficking of E-cadherin and PAR-3from plasma membrane into the cytoplasm with the distance from far to near to tumor focus.6.The model tissues was treated with evodiamine for6h,the mRNA expression of E-cadherin and PAR-3were relatively lower,when treated with24h, the mRNA expression of E-cadherin is relaticvely higher,while the mRNA expression of PAR-3was no obviously variation related to not treated. The model tissues was treated with berberine for6h,the mRNA expression of E-cadherin and PAR-3were relatively lower, when treated with24h, the mRNA expression of PAR-3is relaticvely higher,while the mRNA expression of E-cadherin was no obviously variation related to not treated.7.the model tissues was treated with evodiamine and berberine for6h and24h, The expressions of miR-200a-5p were all relatively higher,while the expression of miR-429was only higher when treated with evodiamine for6h, its expression was lower when treated with berberine fot6h and24h,and treated with evodiamine for24h.Conclusion1.The crypt disorder became worse and the expression of E-cadherin and PAR-3decreased progressively from far to near to tumor focus in the tissues of different distance. 2.Inducing the organization structure change of neonatal rats colon by TGF-β,Then treated with evodiamine and berberine,It is not sure that evodiamine and berberine can reverse the decreasing of E-cadherin and PAR-3expression.