重楼皂苷Ⅰ对人骨肉瘤MG63细胞增殖、凋亡及DNA损伤影响研究  

Study on effect of polyphyllin I on proliferation,apoptosis and DNA damage of human osteosarcoma MG63 cell line

作　　者：王洪伸 林涌鹏[1] 尹萌辰[4] 常君丽[4] 李欣怡[1] 李永津 陈博来[1] WANG Hongshen;LIN Yongpeng;YIN Mengchen;CHANG Junli;LI Xinyi;LI Yongjin;CHEN Bolai(Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510120, Guangdong,China;Postdoctoral Research Station of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;Postdoctoral Research Station of Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510120, Guangdong,China;Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China)

机构地区：[1]广东省中医院,广东广州510120 [2]上海中医药大学博士后科研流动站,上海201203 [3]广东省中医院博士后科研工作站,广东广州510120 [4]上海中医药大学附属龙华医院,上海201203

出　　处：《现代中西医结合杂志》2021年第36期3991-3996,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金青年基金项目(81704097);广州市科技计划项目(202102010012);广东省普通高校特色创新类项目(2019KTSCX024)。

摘　　要：目的探讨重楼皂苷Ⅰ对人骨肉瘤MG63细胞增殖、凋亡及DNA损伤的影响。方法将MG63细胞分为PBS对照组、0.625μmol/L重楼皂苷Ⅰ组、1.25μmol/L重楼皂苷Ⅰ组,各组给予相应干预72 h后,采用Countstar细胞计数仪检测细胞存活率;将MG63细胞分为PBS对照组、0.625μmol/L重楼皂苷Ⅰ组,划痕实验检测各组培养0 h、24 h、48 h、72 h细胞迁移能力;将MG63细胞分为PBS对照组、0.625μmol/L重楼皂苷Ⅰ组、1.25μmol/L重楼皂苷Ⅰ组,流式细胞术检测干预24 h后各组细胞凋亡率;将MG63细胞分为PBS对照组、0.625μmol/L重楼皂苷Ⅰ组、1.25μmol/L重楼皂苷Ⅰ组,TUNEL法和彗星实验分别检测细胞DNA损伤情况;Western blot法检测1.25μmol/L重楼皂苷Ⅰ处理0 h、0.5 h、1.0 h、1.5 h、3 h、6 h后MG63细胞中Bax、Bcl-2、Cleaved PARP蛋白表达情况。结果0.625μmol/L重楼皂苷Ⅰ组和1.25μmol/L重楼皂苷Ⅰ组细胞存活率均明显低于PBS对照组(P均<0.05),且1.25μmol/L重楼皂苷Ⅰ组明显低于0.625μmol/L重楼皂苷Ⅰ组(P<0.05)。培养24 h、48 h、72 h时,0.625μmol/L重楼皂苷Ⅰ组的划痕宽度均大于PBS对照组(P均<0.05)。0.625μmol/L重楼皂苷Ⅰ组和1.25μmol/L重楼皂苷Ⅰ组细胞凋亡率、TUNEL/DPAI荧光值、细胞DNA损伤率均明显高于PBS对照组(P均<0.05),且1.25μmol/L重楼皂苷Ⅰ组均明显高于0.625μmol/L重楼皂苷Ⅰ组(P均<0.05)。1.25μmol/L重楼皂苷Ⅰ培养后0 h、0.5 h、1.0 h、1.5 h、3 h、6 h,Bax、Cleaved PARP蛋白表达随时间延长而逐渐增高,Bcl-2蛋白表达逐渐降低。结论重楼皂苷Ⅰ能够通过诱导DNA损伤抑制骨肉瘤MG63细胞增殖、迁移,促进细胞凋亡,发挥抑制骨肉瘤的作用。Objective It is to investigate the effect of polyphyllin I on MG63 cell line proliferation,apoptosis and DNA damage in human osteosarcoma.Methods The MG63 cells were divided into PBS control group,0.625μmol/L polyphyllin I group,and 1.25μmol/L polyphyllin I group.After 72 hours of intervention in each group,the survival rate of the cells was measured by the Countstar cell counter.The MG63 cells were divided into PBS control group and 0.625μmol/L polyphyllin I group,the cell migration ability of each group cultured for 0 h,24 h,48 h,and 72 h were detected by scratch test.The MG63 cells were divided into PBS control group,0.625μmol/L polyphyllin I group and 1.25μmol/L polyphyllin I group,the apoptosis rate of each group after 24 hours of intervention was detected by flow cytometry.The MG63 cells were divided into PBS control group,0.625μmol/L polyphyllin I group,and 1.25μmol/L polyphyllin I group,the cell DNA damage was detected by TUNEL method and comet assay;the expression of Bax,Bcl-2,Cleaved PARP protein in MG63 cells after treatment with 1.25μmol/L polyphyllin I for 0 h,0.5 h,1.0 h,1.5 h,3 h,and 6 h was detected by Western blot.Results The cell survival rate of 0.625μmol/L polyphyllin I group and 1.25μmol/L polyphyllin I group was significantly lower than that of the PBS control group(all P<0.05),and the cell survival rate of 1.25μmol/L polyphyllin I group was significantly lower than 0.625μmol/L polyphyllin I group(P<0.05).When cultured for 24 h,48 h,72 h,the scratch width of 0.625μmol/L saponin I group was larger than that of PBS control group(all P<0.05).The cell apoptosis rate,TUNEL/DPAI fluorescence value,and cell DNA damage rate of 0.625μmol/L polyphyllin I group and 1.25μmol/L polyphyllin I group were significantly higher than those of the PBS control group(P<0.05),and the rate of the 1.25μmol/L polyphyllin I group was significantly higher than that of the 0.625μmol/L polyphyllin I group(all P<0.05).After culture with 1.25μmol/L polyphyllin I for 0 h,0.5 h,1.0 h,1.5 h,3 h,6 h,Bax and C

关 键 词：骨肉瘤 重楼皂苷Ⅰ DNA损伤 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]