摘要
目的研究二烯丙基二硫(diallyl disulfide,DADS)对人胃癌MGC803细胞G2/M期检查点Chk1与Chk2的影响。方法流式细胞术检测细胞周期改变;Northern blot、Westernblot与免疫细胞化学检测DADS处理MGC803细胞的Chk1与Chk2表达。结果流式细胞术分析显示,30 mg.L-1DADS呈时间依赖性阻滞MGC803细胞在G2/M期(P<0.05);Northern blot检测表明,DADS不同时间作用MGC803细胞后,Chk1与Chk2 mRNA表达与未处理组差异无显著性(P>0.05);免疫细胞化学发现Chk1与Chk2表达与未处理组无明显改变(P>0.05);Western blot DADS在不同时间对MGC803细胞Chk1与Chk2总蛋白表达无改变(P>0.05),而磷酸化的Chk1表达呈时间依赖性增加(P<0.05),但磷酸化的Chk2无明显改变(P>0.05)。结论 DADS阻滞MGC803细胞G2/M期与磷酸化Chk1有关。
Aim To investigate the effects of dialtyl di- sulfide （DADS） on Gz/M check points, Chkl and Chk2, in human gastric cancer MGC803 cells. Meth- ods Flow cytometry was used to determine the change of cell cycle. The expression of Chkland Chk2 in MGC803 cells treated with DADS was detected by Northern blot, immuocytochemistry and Western blot. Results Flow cytometry analysis revealed that MGC803 cells treated by 30 mg·L-1 DADS accumu- lated cells in G2/M phase, exhibiting a time-dependent model （P 〈0.05）. After treated with 30 mg· L-1 DADS for different time points, Northern blot analysis showed the level of Chkl and Chk2 mRNA had no sig- nificant difference with control in MGC803 cells （P 〉0. 05）. It was revealed that the expression of Chkl and Chk2 proteins had no ehange by immuocytoehemistry （P 〉 0.05 ）. Western blot analysis showed that the to- tal proteins of Chkl and Chk2 did not change, howev- er,phosphorylation of Chkl was signifieantly inereased in a time-dependent manner （P 〈 0. 05）, and the ex- pression of phospho-Chk2 had no significant ehange in MGC803 ceils treated by DADS （P 〉 0.05 ）. Conclu- sion DADS can arrest human gastric cancer cells at GJM, which is related to the phosphorylation of Chkl.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2013年第1期79-84,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81102854,31100935)
湖南省教育厅科学研究重点资助项目(No 09A077)
湖南省高校创新平台开放基金资助项目(No 09K074)
