The Inhibitory Effect With Its Mechanism Of Esculetin To The Hepa1-6Cells And Subcutaneously Translanted Tumor Of Hepa1-6in Mice

ObjectiveResearching the inhibitory effect of roliferation invitro to esculetin on themouse hepatoma Hepa1-6cells ang making mouse subcutaneous model tosurvey the inhibitory effect of esculetin to the tumor growth of translantedHepa1-6in mouse.Investigating the inhibitory effect with its mechanism ofesculetin to the mouse of Hepa1-6cells and tumor growth translanted Hea1-6inmouse.Methods（1）Detecting the inhibitory effect of different concentrations of esculetin anddifferent rocessing time on Hea1-6cells by using MTT assay.FCM was used toassay the change of aototic rate of different concentrations of esculetin onHea1-6cells.（2）Hepa1-6cells line （murine） at37percent degree celsius,5%C02incubator.The Hepa1-6tumor cells were inoculated in the right front armitits ofthe mices for making mice subcutaneous model.40mices were allocated intofollowing5groups randomly: Saline group0.2ml·d^-1, cyclohoshamide（CTX）group100mg·kg·d^-1, Esculetin High dose group700mg·kg·d^-1,med-ium dose group400mg·kg·d^-1,low dose group200mg·kg·d^-1.The third days after intraeritoneal injection to medicine,the mouse were weighted every twodays.The mouse were killed after21days and tumor tissues searated andweighted;Detecting the expression of mRNA of aotosis genes Bax ang Bcl-2byusing the technology of RT-CR;detecting the protein expression of Bax,Bcl-2and VEGF by using the technology of Western blot.Results（1） The effect of roliferation in vitro to esculetin on the Hea1-6cells:thedifferent inhibitory effect of5concentrations of esculetin to Hea1-6cells by usingMTT assay showed that the concentrations of esculetin were1.04mmol·L^-1,2.08mmol·L^-1and4.16mmol·L^-1,which have an effect on roliferation;the result ofFCM showed that with the increasing of the concentrations of esculetin theaotosis was raised;esculetin may disturb the normal cell cycle ang make thecells in the stage of S.（2） From the seventh day,the mouse of model group became thin and theeleventh day the mouse of cyclophosphamide group became thin too they wereanorexia and insensitive until the end of the experiment.But the mouse ofgroups of esculetin were not obvious;The effect of esculetin to the mice ofsubcutaneously translanted tumor:the inhibition ratio of esc-uletin high,medium and low dose groups were55.42%,40.37%and,coma-ring with model group the differences were statistical significance （P﹤0.01）;upregulateding Bax rotein expression while downregulateding Bcl-2roteinexpression, the extent of expression relate to the dose,the rotein expression ofVEGF depend on the dose,too;Comared to the model group,the expression ofeach group of esculetin Bax gene mRNA and Bcl-2gene mRNA.The differencewas statistically significant （P﹤0.05）. ConclusionsThe esculetin can make certain inhibition effect to the growth of Hepa1-6cells and thetumor translanted with Hepa1-6; the effect of esc-uletin on Hepa1-6cells is concerned with motivateing cell roliferation. Itsmechanism is robably raise the expression of Bax rotein genes and reduce theexpression of Bcl-2rotein genes.and also concerned with inhibiting tumorvascular endothelial growth factor expression of VEGF.