褪黑素对骨肉瘤MG-63细胞的增殖和抑制作用的体外实验研究

Effects of Proliferation and Inhibition of Melatonin on Growth of Human Osteosarcoma Cell Line MG-63 Cell

目的:研究褪黑素对人成骨肉瘤细胞株MG-63增殖和抑制作用。方法:不同浓度的褪黑素作用于体外培养的MG-63细胞,倒置显微镜下观察MG-63细胞的形态学变化,采用四甲基偶氮唑蓝(MTT)法检测不同浓度褪黑素对MG-63存活率,细胞计数试剂8(CCK-8)法检测褪黑素对MG-63细胞增殖抑制能力,Annexin V/PI双染法标记检测细胞凋亡情况以及黏附实验测定细胞黏附能力和培养液中LDH释放量的检测。结果:MTT法检测经褪黑素作用后的MG-63细胞存活率以及细胞黏附能力明显下降,LDH的增加提示MG-63细胞死亡率上升;褪黑素可抑制骨肉瘤细胞株MG-63细胞增殖,5mmol/L的褪黑素对MG-63细胞诱导凋亡作用最明显,不同浓度的褪黑素之间实验结果具有统计学意义。结论:褪黑素能够诱导MG-63细胞凋亡,降低存活率,抑制细胞增殖,是治疗骨肉瘤潜在的靶向药物。

Objective： To investigate the effects ofmelatonin （MLT） on proliferation and apoptosis effect of human osteosarcoma cell MG-63. Methods： The cultured MG-63 cells were treated with MLT of different concentrations, and the morphological changes were observed under inverted microscope. The survival ratio of MG-63 was assessed through the 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenylte- trazolium bromide （MTT） assay and the inhibitory rates of MG-63 was detected through cell counting ofkit-8（CCK-8） assay. The apoptosis MG-63 was measured through double staining of Annexin V/PI, the adhesion capacity of MG-63 and releasing of lactate dehydrogenase （LDH） were evaluated through adhesion assay. Results： The survival ratio and adhesion capacity of MG-63 decreased significantly after treatment with different concentrations of MLT, and the releasing of LDH was increased, implicating the morality rate of MG-63 was ele- vated. The MLT could inhibit the proliferation of MG-63, with the maximal effects of induction of apoptosis at concentrations of 5mmol/L, there also existed statistical significance among treatments with different concentrations of MLT. Conclusions： MLT could in- duce the apoptosis of MG-63, reduce its survival rate and inhibit its proliferation, and thus could be a potential targeted drug for treatment of osteosarcoma.