蝎毒多肽对人骨肉瘤细胞株MG-63活性的影响  

Effect of scorpion venom polypeptide on the activity of human osteosarcoma cell line MG-63

作　　者：徐绘华[1] 罗仕勇 黄智通 郭奇峰[1] XU Hui-hua;LUO Shi-yong;HUANG Zhi-tong;GUO Qi-feng(Department of Orthopedics,Guangzhou First People′s Hospital,Guangzhou 510180,China)

机构地区：[1]广州市第一人民医院骨科,广东广州510180

出　　处：《解剖学研究》2020年第2期112-115,123,共5页Anatomy Research

基　　金：国家自然科学基金资助项目(31601031)

摘　　要：目的探讨蝎毒多肽对人骨肉瘤MG-63细胞增殖凋亡的影响。方法利用0、4、8、12μg/mL浓度蝎毒多肽处理MG-63细胞12、24、48 h,CCK-8法检测细胞活性、镜下观察细胞形态变化、吖啶橙荧光染色法观察细胞凋亡,8μg/mL蝎毒多肽处理MG-63细胞24、48 h,Hoechst-33258荧光染色观察细胞核染色质形态。结果 4、8、12μg/mL的蝎毒多肽可抑制MG-63细胞活性,抑制其增殖,呈剂量-时间依赖性。8μg/mL处理24 h细胞存活率为0.5506±0.0502,与对照组比较,差异有统计学意义(P<0.01)。吖啶橙荧光染色,细胞染色质浓缩,细胞核碎裂成点状,被染成大小不一、致密浓染的绿色颗粒,随着浓度增大、时间延长,坏死细胞逐渐增多。呈现凋亡核碎裂典型变化,随剂量增加更趋明显。呈剂量依赖性抑制其增殖能力。Hoechst-33258染色观察可见凋亡细胞染色质呈固缩或碎裂状染色质,染色质着色不均,核形态各异。随作用时间增加而更趋明显。结论蝎毒多肽可以抑制骨肉瘤细胞活性,抑制其体外增殖,诱导其发生凋亡,该作用呈剂量依赖和时间依赖。Objective To investigate the effect of scorpion venom polypeptide on the proliferation and apoptosis of human osteosarcoma MG-63 cell line.Methods MG-63 cells were treated with 0,4,8,12 μg/mL scorpion venom polypeptide for 12,24,48 hours,then the activity of cells was detected by CCK-8 method,the morphological changes were observed under microscope,the apoptosis was observed by acridine orange fluorescence staining,and the morphology of MG-63 cells was observed by hoechst-33258 fluorescence staining after 24,48 hours.Results 4,8,12 μg/l scorpion venom polypeptide could inhibit the activity and proliferation of MG-63 cells in a dose-time dependent manner.The cell survival rate was 0.5506±0.0502 after 24 hours treatment with 8 μg/mL.The difference was statistically significant compared with the control group(P <0.01).After treatment with different concentrations of drugs,acridine orange fluorescence staining,chromatin concentration,nuclear fragmentation into dots,dyed into green particles of different sizes and dense concentration.With the increase of concentration and time,necrotic cells gradually increased.The cells showed typical changes,such as apoptotic nuclear fragmentation,which became more obvious with the increase of drug dosage.It inhibited the proliferation in vitro in a dose-dependent manner.Hoechst-33258 staining showed that the chromatin of apoptotic cells was white,pyknosis or fragmentation,the chromatin was not uniform,and the nuclear morphology was different.Conclusion Scorpion venom polypeptide can inhibit the activity of osteosarcoma cells,inhibit their proliferation in vitro and induce their apoptosis,which is dose-dependent and time-dependent.

关 键 词：蝎毒多肽 人骨肉瘤MG-63细胞 增殖 凋亡 

分 类 号：R738[医药卫生—肿瘤]