CAJ | 학술논문

目的以抑癌基因即腺瘤性结肠息肉病相关基因(APE)存在异常甲基化的急性T淋巴细胞白血病Jurkat细胞为研究对象,探究雷公藤内酯醇(TPL)对APC基因的影响,并对其机制进行初步探讨.方法运用四甲基偶氮唑盐(MTT)法等检测TPL对Jurkat细胞株增生的影响.巢式甲基特异性PCR(n-MSP)检测TPL对Jurkat细胞APC基因甲基化模式的影响.半定量RT-PCR检测经TPL作用后Jurkat细胞APC基因、甲基转移酶DNMT3A、DNMT3B mRNA的表达.Western blotting检测TPL作用后APC蛋白的表达.结果 TPL对Jurkat细胞生长有明显的抑制作用,并有时间及剂量依赖性,48 h IC50为19.7 ng/ml;TPL可逆转APC基因的高甲基化;TPL能够诱导Jurkat细胞APC基因mRNA的表达,具剂量依赖性;TPL能够诱导Jurkat细胞APC蛋白重新表达,亦有剂量依赖性.结论小剂量TPL可明显抑制Jurkat细胞的增生,其可能机制为通过诱导Jurkat细胞中异常甲基化的APC基因去甲基化,使APC基因恢复表达.
목적 이억암기인즉선류성결장식육병상관기인(APE)존재이상갑기화적급성T림파세포백혈병Jurkat세포위연구대상,탐구뢰공등내지순(TPL)대APC기인적영향,병대기궤제진행초보탐토.방법 운용사갑기우담서염(MTT)법등검측TPL대Jurkat세포주증생적영향.소식갑기특이성PCR(n-MSP)검측TPL대Jurkat세포APC기인갑기화모식적영향.반정량RT-PCR검측경TPL작용후Jurkat세포APC기인、갑기전이매DNMT3A、DNMT3B mRNA적표체.Western blotting검측TPL작용후APC단백적표체.결과 TPL대Jurkat세포생장유명현적억제작용,병유시간급제량의뢰성,48 h IC50위19.7 ng/ml;TPL가역전APC기인적고갑기화;TPL능구유도Jurkat세포APC기인mRNA적표체,구제량의뢰성;TPL능구유도Jurkat세포APC단백중신표체,역유제량의뢰성.결론 소제량TPL가명현억제Jurkat세포적증생,기가능궤제위통과유도Jurkat세포중이상갑기화적APC기인거갑기화,사APC기인회복표체.
Objective To observe the effects of triptolide(TPL) on the anti-oncogene-APC gene of acute lymphoblastic leukemia cell line Jurkat in vitro. Methods The effects of TPL on proliferation Jurkat cells were assayed by using cell culture, MTT. The effects of TPL on APC gene of Jurkat cells were analyzed by nested methylation specific PCR and RT-PCR. The effects of TPL on the proteinum expression of APC gene were detected by Western blotting analysis. Results Following the treatment of TPL, the cell proliferation rate was degraded as the treatment concentration increased and the culture time extended. The effects were dose and time-dependent. The 48 hour IC50 was 19.7 ng/ml. TPL can reverse hypermethylation of APC gene,and induce the expression of the mRNA and the proteinum. Conclusion Low dose TPL could depress the proliferation rate of Jurkat. The possible mechanism might be its reversing the hypermethylation of APC gene and activiting the expression of APC gene.