摘要
目的探索重组天花粉蛋白(recombinant trichosanthin,rTCS)高表达对人宫颈癌Caski细胞p73基因甲基化影响的机制。方法 RT-PCR、QT-PCR、Western blot检测rTCS高表达后Caski细胞中DNA甲基化转移酶I(DNMT1)的表达变化,甲基化特异性PCR(MSP)法检测p73基因5'端启动子区CpG岛甲基化状态的改变。结果宫颈癌Caski细胞p73为低表达,其启动子呈部分甲基化状态。高表达rTCS的细胞株其DNMT1mRNA及蛋白水平的表达均降低,其中mRNA水平降低0.56倍(P<0.01),p73基因启动子甲基化程度亦有所降低,p73mRNA水平表达升高。结论高表达rTCS可能通过抑制DNMT1的表达,致使抑癌基因p73启动子去甲基化而重新表达,最终发挥抑癌基因功能抑制宫颈癌Caski细胞增殖。
Aim To study the effects of high expression of recombinant trichosanthin （ rTCS ） on p73 methyla- tion and mechanism research in Caski cell. Methods RT-PCR, QT-PCR, Western blot were used to deter- mine DNMT1 expression. The methylation status of p73 promoter was detected by Methylation PCR. Re- suits DNMT1 expression in mRNA and protein levels was lower in the rTCS high expression cells than in the control cells, with a 0. 56-fold decrease for mRNA （P 〈 0.01 ）. The methylation level of p73 promoter in theexperimental group was lower than the control cells. p73 expression in the experimental group was higher than the control cells in mRNA level. Conclusion The high expression of rTCS inhibits Caski cell prolifer- ation through suppressing the expression of DNMT1, which results in demethylation of p73 promoter and p73＇s re-expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2013年第9期1290-1293,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 30873282)
湖北省卫生厅2011年年度科研资助项目(No NX2011-9)
