Studies On Induced Apoptosis Of K562,NCI-H460 Cells And Inhibition Of Transplanted Tumor Growth In Nude Mice By Anthocyanin

Objective: To study the effect induced apoptosis of K562,NCI-H460 cells by anthocyanidin.Methods: K562,NCI-H460 cells were cultured with different concentrations of anthocyanidin (25,50,100 and 200μg/ml) in vitro, the growth inhibition effect and the IC50 value were assayed by MTT method. Cell viability was analyzed by trypan blue exclusion assay. Morpho1ogica1 changes of cells were observed with microscopy. The typical DNA ladder was detected by DNA agarose gel electrophoresis. Morphology of cells nucleus apoptosis were observed by Hoechst 33258 fluorescent staining. Cell apoptosis rate and cell cycle distribution were further analyzed by flow cytometry(FCM).Results: K562 and NCI-H460 cells viability decreased obviously and presented different morphological changes by concentrations anthocyanidin between 25 to 200μg/ml. The proliferation inhibitory ratio of K562 and NCI-H460 cells after treated by anthocyanidin (25,50,100 and 200μg/ml)for 72 hours were 16.0,33.6,53.9,65.1 and 23.0,38.1,50.6,59.1. The IC50 values of anthocyanidin on K562 and NCI-H460 cells were 89.1,90.8μg/ml. K562 and NCI-H460 cells proliferation were remarkably inhibited by different concentration between 25 to 200μg/ml and the inhibition was both in-dose and time-dependent manner. K562 and NCI-H460 cells were exposed to anthocyanidin (50～200μg/ml ) for 72 hours, the typical DNA ladder on agarose gel electrophoresis for analysis of cellular apoptosis was significantly appeared, marked nuclear shrink and nuclear fragmentation morphological changes of cells apoptosis were observed by Hoechst 33258 fluorescent staining. Simultaneously, the sub-G1 peak was found in FCM analysis, the proportion of K562 and NCI-H460 cells in G0/G1-phase of cell cycle was significantly increased and the proportion cells in S-phase was decreased with the increasing concentration of anthocyanidin. The apoptosis rate of K562 and NCI-H460 cells were 57.4,27.8% by 200μg/ml anthocyanidin.Conclusions: Anthocyanidin(25～200μg/ml)can inhibit proliferation of K562,NCI-H460 cells and the inhibition was both in-dose and time-dependent manner. Anthocyanidin(50～200μg/ml)can induce apoptosis of K562,NCI-H460 cells in dose-dependent manner and cause cells detained at G0/G1 phase. Induction apoptosis of K562,NCI-H460 cells is one of the anti-tumor mechanisms of anthocyanins.2. The study on inhibition of tumor growth in K562,NCI-H460 cells tumor-bearing nude mice by anthocyanidinObjective: To investigate the inhibitory effect of anthocyanidin on tumor growth in k562,NCI-H460 cells tumor-bearing mice in vivo.Methods: K562,NCI-H460 cells and tumor block were subcutaneously injected into nude mice to establish model of transplanted tumor. Nude mice were randomly divided into model group,the treatment group and the positive control group. Model group was given normal sodium 20 ml/(kg.d), the treatment group was given to 50,100 and 150 mg/(kg.d) anthocyanins gavage administration 15 days, the positive control group was given 20 mg/ kg every two days cyclophosphamide gavage administration 8 days. The nude mice were continued to be raised for one month after stopping the medicine. The kinetics of tumor formation and tumor growth were measured, tumor volume was the survey, tumor growth curve was drawn, tumor growth inhibition rate (IR) was calculated and the survival situation of nude mice bearying-tumor was observed.Results: The K562 cell line and the lump block's nude mice transplant lump success ratio is extremely low. The NCI-H460 cell line and the lump block's transplant success ratio reaches 100%. The anthocyanidin has the obvious inhibitory action to the NCI-H460 bare mouse transplant lump's growth. The middle and high dose group inhibitory rate reach to 47.1% and 58.9% in end time giving the medicine. The bare mouse beard lump survival situation is bad, the state of mind is bad, slow-acting, becomes emaciated obviously.Conclusions: The K562 cell line and the lump block's nude mice transplant lump success ratio is extremely low. The NCI-H460 cell line and the lump block's transplant success ratio reaches 100%. The anthocyanidin has the obvious inhibitory action to the NCI-H460 bare mouse transplant lump's growth. The anthocyanidin has certain prevention function to the NCI-H460 cell bare mouse transplant lump's growth.