A study of combined killing effects of cinobufagin and cisplatin on osteosarcoma MG-63 cells

SONG Ke-guan; ZHANG Tao; YAN Feng; SUN Yuan-xin; WANG Hai; ZHOU Yong-fei

doi:10.3969/j.issn.2095-252X.2014.03.016

Abstract：Objective To investigate and compare killing effects of cinobufagin and cisplatin on human osteosarcoma MG-63 cell lines. Methods Osteosarcoma MG-63 cell lines were cultured in vitro, and logarithmic growth phase cells were selected for experiments. The methyl thiazolyl tetrazolium ( MTT ) colorimetry assay was used to determine the growth inhibitory effects on MG-63 cells and to calculate the cell inhibitory rate. The flow cytometry was used to detect the apoptosis rate and cell cycle of osteosarcoma cells, and to observe the changes of cell apoptosis rates and the inhibitory effects of drugs on cell cycles. The in situ terminal deoxynucleotidyl transferase ( TdT )-mediated dUTP-X nick end labeling ( TUNEL ) was used to observe the apoptosis rate of osteosarcoma cells and to detect the cell apoptosis rate. The osteosarcoma cell growth status and morphological changes were observed by the phase contrast microscope. Results Both cinobufagin and cisplatin could inhibit the growth of osteosarcoma cells. In the cinobufagin group, cisplatin group and cinobufagin+cisplatin group, the inhibitory effects on the osteosarcoma cell growth were not identical at different drug concentrations. When the drug concentration was less than 80 μg/ml, the cell growth inhibitory effects in all experimental groups would become greater with the increase of drug concentration. When the drug concentration reached 80 μg/ml, the cell growth inhibitory effects in the cinobufagin group and cisplatin group would not increase. And when the drug concentration reached 160 μg/ml, the cell growth inhibitory effects in the cinobufagin+cisplatin group were greater than the effects at the concentration of 80 μg/ml. In all experimental groups, the growth inhibitory effects of drugs at the same concentration on osteosarcoma cells in the cinobufagin+cisplatin group were greater than that in the single cinobufagin or cisplatin group. Conclusions The combined killing effects of cinobufagin+cisplatin on osteosarcoma MG-63 cells are obviously greater than that of the single cisplatin or cinobufagin. In the inhibition of osteosarcoma MG-63 cell growth, the cell killing effects of cinobufagin combined with cisplatin are signiifcant, with a time-and concentration-dependence.