CAJ | 학술논문

目的研究异硫氰酸苯己酯(PHI)对急性T淋巴细胞性白血病Molt-4细胞p15基因的去甲基化作用及转录激活作用.方法采用甲基特异性聚合酶链反应(MSP)检测PHI作用前后Molt-4细胞株p15基因甲基化状态的变化情况;RT-PCR检测p15基因的mRNA的表达变化;Western blotting检测p15蛋白的表达变化.结果 PHI作用于Molt-4细胞5 d后,p15基因的异常甲基化现象被逆转,基因的甲基化程度减弱;基因转录激活,p15 mRNA、p15蛋白表达呈浓度依赖性增加.各组p15 mRNA条带灰度值与β-actin比值为:空白对照组(0.17±0.12),PHI 10 μmol/L组(0.29±0.14),PHI 20 μmol/L组(0.55±0.07),PHI 40 μmol/L组(0.93±0.13),各加药组与空白对照组相比,差异均有统计学意义(P<0.05).结论 PHI有DNA去甲基化的作用,能诱导沉默的p15基因重新表达.
목적 연구이류청산분기지(PHI)대급성T림파세포성백혈병Molt-4세포p15기인적거갑기화작용급전록격활작용.방법 채용갑기특이성취합매련반응(MSP)검측PHI작용전후Molt-4세포주p15기인갑기화상태적변화정황;RT-PCR검측p15기인적mRNA적표체변화;Western blotting검측p15단백적표체변화.결과 PHI작용우Molt-4세포5 d후,p15기인적이상갑기화현상피역전,기인적갑기화정도감약;기인전록격활,p15 mRNA、p15단백표체정농도의뢰성증가.각조p15 mRNA조대회도치여β-actin비치위:공백대조조(0.17±0.12),PHI 10 μmol/L조(0.29±0.14),PHI 20 μmol/L조(0.55±0.07),PHI 40 μmol/L조(0.93±0.13),각가약조여공백대조조상비,차이균유통계학의의(P<0.05).결론 PHI유DNA거갑기화적작용,능유도침묵적p15기인중신표체.
Objective To investigate the effect of phenylhexyle isothiocyanate (PHI) on demethylation and activation of transcription gene p15 in acute leukemia cell line Molt-4. Methods DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Moh-4 cells were treated with PHI. P15 mRNA was measured by RT-PCR. Pl5 protein was detected by Western blotting. Results Hypermethylation of gene pl5 was apparently attenuated and activation of transcription p15 gene was de novo after 5 days exposure to PHI. PHI enhanced both the expression of p15 mRNA and p15 protein in a concentration-dependent manner. The ratio of the gray scale of p15 mRNA strap was 0.17±0.12 in control, 0.29±0.14 in PHI 10 μmol/L, 0.55±0.07 in PHI 20 μmol/L, 0.93±0.13 in PHI 40 μmol/L. Conclusion PHI could active demethylation and transcription of gene p15.