摘要
目的探讨mytoxin B和myrothecine A对肝癌细胞SMMC-7721的增殖抑制作用及分子机制。方法采用MTT法检测细胞增殖,流式细胞术检测细胞周期变化,Western blot检测细胞周期负调控蛋白p27蛋白的表达水平及细胞转染pSi-p27-469质粒(pSi-p27),以观察mytoxin B和myrothecine A是否通过调节p27蛋白表达影响SMMC-7721细胞的增殖。结果 mytoxin B和myrothecine A对SMMC-7721细胞增殖皆有较强的抑制作用,且myrothecine A呈现出明显的剂量依赖性;其增殖抑制作用与S期细胞周期阻滞有关。进一步研究发现,mytoxin B和myrothecine A皆可导致p27蛋白表达明显增加;而SMMC-7721细胞在转染pSip27 48 h后,p27蛋白的表达明显减少;同时,myrothecine A所致的细胞增殖抑制完全被阻断,而mytoxin B所致的细胞增殖抑制部分被逆转。结论 myrothecine A主要通过增加p27蛋白的表达而抑制SMMC-7721细胞增殖,而mytoxin B对SMMC-7721细胞的增殖抑制作用除与上调p27蛋白表达有关外,可能还有其他相关作用机制。
Objective To investigate the molecular mechanism of mytoxin B and myrothecine A in inhibiting the proliferation of SMMC-7721 of hepatocarcinoma cells.Methods MTT assay,flow cytometric analysis,westem blot and pSi-p27-469 plasmid （pSi-p27） cell transfection experiment were used to detect the proliferation activity,cell cycle distribution,expression of negative cell cycle regulatory protein p27 and influence of p27 on cell proliferation.Results The results showed that mytoxin B and myrothecine A exerted potent anti-proliferative activity on SMMC-7721 cells and myrothecine A presented dependent relation.The proliferation inhibition activity of the two compounds were both related to S phase delay.Further research indicated that both mytoxin B and myrothecine A could increase the expression of protein p27 obviously.The pSi-p27 cell transfection experiment discovered that the expression of protein p27 decreased after 48 hours and proliferation inhibition of myrothecine A was blocked completely,and proliferation inhibition of mytoxin B was rescued partly.Conclusion It is proved that myrothecine A can inhibit proliferation of SMMC-7 7 2 1 cells through increasing the expression level of protein p27,while there are other mechanisms involved in proliferation inhibition induced by mytoxin B in SMMC-7721 cells as well as protein p27.
出处
《实用临床医药杂志》
CAS
2014年第11期1-5,共5页
Journal of Clinical Medicine in Practice
基金
国家自然科学基金项目(21372191)
2012年国家级大学生创新创业训练计划(201211117022)
2012年江苏省大学生实践创新训练计划项目(2012JSSPITP1334)
