Synergi Stic Inhibitory Effect Of Evodiamine Combined With Chemotherapy On Hepatocellular Carcinoma Cells

ObjectiveAt present,malignant tumor is the leading cause of death,especially liver cancer in China.Our previous studies have found that both berberine and evodiamine,the main components of ZuoJinFang,have antitumor effects.Evodiamine can inhibit the proliferation of tumor cells by blocking their cell cycle.Other studies have found that evodiamine has an restraining effect on the invasion and metastasis of liver cancer cells.On this basis,we will further focus on the effect of the main components of ZuoJinFang combined with chemotherapy on the activity of hepatocellular carcinoma cells.In vivo and in vitro experiments were conducted to explore whether there is synergistic effect of evodiamine combined with the chemotherapy drug paclitaxel on the growth of hepatocellular carcinoma cells,so as to provide some theoretical basis for future clinical use.MethodsIn vitro culture of human hepatocellular carcinoma cells HepG2,the effect of the combination of evodiamine and chemotherapy on the growth and proliferation of hepatocellular carcinoma cells for 48H under different concentrations was detected by MTT method and High content screening.The effect of evodiamine combined with paclitaxel on the proliferation of hepatocellular carcinoma cells for 48H was detected by plate cloning formation experiment.Flow cytometry was used to detect the effect of evodiamine combined with paclitaxel on the apoptosis rate for 48H.Mouse H22 cells were cultured by passage of ascites tumor,and the growth inhibition of hepatocellular carcinoma cells by combination of evodiamine and chemotherapeutic drugs was detected by mouse underarm tumor transplantation model.Results(1)The MTT method and the High Content screening showed that the combination of evodiamine with paclitaxel or pentafluouracil had a growth inhibition effect on the HepG2 cells in a concentration-dependent way.Most of the combined groups had a synergistic effect with Q larger than 1.15.There was no synergism of evodiamine combined with cisplatin.(2)The plate cloning formation test detected that the colony of cloned cells in the combination of evodiamine plus paclitaxel group was significantly less than that in the control group and the single drug group,and it had an inhibitory effect on the proliferation of hepatocellular carcinoma cells,compared with that in the single drug group,with a statistical difference of P<0.05.The Q value is 1.323,indicating that the combination has synergistic effect.(3)Flow cytometry detection of the effect of evodiamine combined with paclitaxel for 48H showed that the rate of apoptotic cells in the combined group was significantly increased,compared with that in the single drug group,with a statistical difference of P<0.05.The Q value is 1.561,indicating that the combination has synergistic effect.(4)It was found that the tumor volume and weight of the experimental groups were reduced by different degrees through the model of axillary tumor transplantation in mice.The group of evodiamine combined with paclitaxel was significantly lower than that of the control group and the group of evodiamine or paclitaxel monotherapy,with a statistical difference of P<0.05.ConclusionThrough evodiamine combined with paclitaxel on the growth proliferation and apoptosis of hepatocellular carcinoma cells in vivo and in vitro experiments showed that,evodiamine combined with commonly used chemotherapy paclitaxel had an concentration dependently inhibitory effect on the growth of hepatocellular carcinoma cells.In vivo and in vitro experiments showed that evodiamine combined with first-line chemotherapy paclitaxel could synergisticly inhibit the growth and proliferation of hepatocellular carcinoma cells and induced the apoptosis of tumor cells.