Study On Effects Of Yiqiyangyinjiedufang On Proliferation And Apoptosis Of Mouse Lewis Lung Cancer Cells

Objective: (1) Observed the effects of proliferation and apoptosis on Lewis lung cancer cells by yiqiyangyinjiedufang; (2) Observed the effect of yiqiyangyinjiedufang on proliferation of Lewis lung transplanted tumor and apoptosis of Lewis lung transplanted tumor cells; (3)Detected the effect of yiqiyangyinjiedufang on the expression of Caspase-3, preliminarily study the mechanism of yiqiyangyinjiedufang on tumor.Methods:(1)The contained drug serum of low, medium, high dose Yiqiyangyin jiedufang and Cisplatin(DDP), Yiqiyangyinjiedufang jointed DDP, normal saline(NS) was taken from SD rats and was inactivated before the experiment on the base of the serum pharmacology method, then the serum was added to LLC in vitro, the proliferation of LLC was measured by methyl thiazolyl tetrazolium(MTT) method.(2) Establish the Lewis lung carcinoma models of C57BL/6 mice, sixty C57BL/6 mice were randomly divided into 6 groups: control group, yiqiyangyinjiedufang low dose group, medium dose group, high dose group, DDP group, yiqiyangyinjiedufang combine with DDP group. 24 hours after tumor implantation, every group of yiqiyangyinjiedufang and NS group were administered lavage with yiqiyangyin jiedufang and NS for 20 days, DDP group and combine group were given DDP once every other day by intraperitoneal injection for 7 days, Volume of tumors were measure respectively during the therapy time, all mice were sacrificed after 28 days since tumors were transplantation, Weigh of tumor and calculate inhibitory rate. (3) The cell apoptosis were measured by flow cytometry(FCM) and Annexin V- FITC/PI, Immunohistochemistry (SP method) measured the expression levels of caspase-3. (4)Assessed apoptosis index of tumor cells by The TUNEL method, SP method measured the expression levels of caspase-3 of Lewis lung transplanted tumor.Results: (1)Yiqiyangyinjiedufang Contained drug serum had a remarkable inhibition on LLC in vitro, these effects showed characters being time dependent and concentration dependent, effect of high concentrations on LLC in 72h, the growth inhibiting rate(IR) was 50%, inhibiting rate of yiqiyangyinjiedufang joint DDP Containing drug serum up to 94.67%. (2)The tumor growth was suppressed significantly in yiqiyangyinjiedufang low dose group, medium dose group, high dose group, the weights of tumor were decreased in every yiqiyangyinjiedufang group compared with control group, the inhibition rates were 19.30%, 32.83% and 34.09%, the difference was significant (P<0.01), Yiqiyangyinjiedufang joint DDP group was 83.71%, compared with the other groups, the difference was significant (P<0.01). (3) Compared with the control group, each group containing serum can induce apoptosis of LLC, and showed characters being time-dependent and concentration-dependent. yiqiyangyinjiedufang joint DDP Containing drug serum on LLC for 72h, the apoptosis rate declined and the necrosis increase, the expression of caspase-3 was being dose an time dependent manner(P<0.05), Compared with other groups, expression of caspase-3 of Chinese combined DDP was highest(P<0.05). (4) Compared with control group, the apoptosis index of Lewis lung transplanted tumor in every yiqiyangyinjiedufang group and yiqiyangyinjiedufang joint DDP group were heighten by TUNEL method, the expression of Caspase-3 in group F was strongest, compared with other groups, the difference was significant(P<0.05), yiqiyangyinjiedufang can induce apoptosis of tumor cells, when joint with DDP, the effect increased(P<0.05), the expression of caspase-3 of every yiqiyangyinjiedufang group was higher than control group, Compared with other groups, expression of caspase-3 of Chinese combined DDP was highest(P<0.05).Conclusion: yiqiyangyinjiedufang can inhibit proliferation and induce apoptosis of LLC, and joint DDP has synergistic effect. The promotion of caspase-3 protein expression may be one of the mechanisms of inducing apoptosis of LLC. Yiqiyangyinjiedufang can inhibit the growth of Lewis lung carcinoma transplanted in mice, when combined with DDP the growth inhibition was increased, and its mechanisms might be relative to promoting the expression of Caspase-3 and inducing apoptosis.