CAJ | 학술논문

目的：探讨新型DNA甲基转移酶（ DNMT）抑制剂SGI-1027对尤文肉瘤SK-N-MC和SK-ES-1细胞系增殖和凋亡的影响。方法：细胞活性分析测定细胞增殖状况。流式细胞计数法测量细胞周期的改变。实时定量PCR和Western blot检测DNMT1、DNMT3a和DNMT3b以及抑癌基因p16、p21的mRNA和蛋白水平的变化以及凋亡剪切产物cleaved PARP的表达。结果：SGI-1027明显抑制尤文肉瘤细胞增殖，诱导G2期细胞增加并可见subG1峰，cleaved PARP表达增加。DNMT1、DNMT3a和DNMT3b的mRNA水平不变，但DN-MT1蛋白水平明显下调；p16和p21的mRNA和蛋白水平上调。结论：新型DNA甲基转移酶抑制剂SGI-1027能够抑制DNMT1活性，激活抑癌基因p16和p21，发挥对尤文肉瘤细胞凋亡诱导作用。
목적：탐토신형DNA갑기전이매（ DNMT）억제제SGI-1027대우문육류SK-N-MC화SK-ES-1세포계증식화조망적영향。방법：세포활성분석측정세포증식상황。류식세포계수법측량세포주기적개변。실시정량PCR화Western blot검측DNMT1、DNMT3a화DNMT3b이급억암기인p16、p21적mRNA화단백수평적변화이급조망전절산물cleaved PARP적표체。결과：SGI-1027명현억제우문육류세포증식，유도G2기세포증가병가견subG1봉，cleaved PARP표체증가。DNMT1、DNMT3a화DNMT3b적mRNA수평불변，단DN-MT1단백수평명현하조；p16화p21적mRNA화단백수평상조。결론：신형DNA갑기전이매억제제SGI-1027능구억제DNMT1활성，격활억암기인p16화p21，발휘대우문육류세포조망유도작용。
Objective:To investigate the anti-proliferative effect of novel DNA methyltransferase( DNMT)inhibi-tor SGI-1027 in Ewingˊs sarcoma cell lines SK-N-MC and SK-ES-1 in vitro. Methods:The proliferation of SK-N-MC and SK-ES-1 cells was determined by cell viability assay after treatment of SGI-1027. Cell cycle dis-tribution was determined by flow cytometry. The mRNA level and protein expression of DNMT1,DNMT3a,DNMT3b, tumor suppressor p16,p21 and production of cleaved PARP were detected by real-time PCR and Western blot,re-spectively. Results:SGI-1027 significantly inhibited cell proliferation and resulted in G2 arrest as well as appearance of subG1 peaK. Western blot showed the expression of cleaved PARP. Although there were no change of mRNA level in DNMT1,DNMT3a and DNMT3b,protein level of DNMT1 was downregulated. The mRNA and protein level of p21 and p16 was upregulated. Conclusion:Novel DNA methyltransferase inhibitor SGI-1027 can inhibit DNMT1 activity and reactivate tumor suppressor p21 and p16,finally induce apoptosis in Ewingˊs sarcoma cell lines.