CAJ | 학술논문

目的探讨三氧化二砷(AS2O3)对多发性骨髓瘤(MM)细胞内细胞因子信号转导抑制因子-1(SOCS-1)基因甲基化状态的影响及其对磷酸化的信号转导与转录激活因子-3(P-STAT3)表达的影响.方法采用甲基特异性PCR法检测AS2O3作用前后MM细胞株U266和CZ-1细胞内SOCS-1基因的甲基化状态,应用蛋白免疫印迹法检测AS2O3处理前后细胞内P-STAT3蛋白的表达变化,并采用流式细胞技术检测AS2O3作用前后MM细胞增殖和凋亡的变化.结果 MM细胞株内SOCS-1基因存在程度不同的甲基化状态,与对照组相比,AS2O3作用后MM细胞内SOCS-1基因甲基化程度明显减弱或消失,P-STAT3蛋白的表达也明显减弱,同时细胞生长受抑,凋亡比率升高.AS2O3浓度分别为0、0.5、1.0、2.0μmol/L时,U266细胞株的总凋亡率分别为0.06％、0.56％、48.96％、61.07％(x2=9.19,P＜0.05)；而CZ-1细胞株的总凋亡率分别为4.20％、40.30％、47.72％、68.49％(x2=8.96,P＜0.05).结论AS2O3可能通过诱导MM细胞内SOCS-1基因去甲基化作用,进一步抑制细胞增殖信号Janus激酶(JAK) -STAT通路的活化,从而诱导MM细胞的凋亡.
목적 탐토삼양화이신(AS2O3)대다발성골수류(MM)세포내세포인자신호전도억제인자-1(SOCS-1)기인갑기화상태적영향급기대린산화적신호전도여전록격활인자-3(P-STAT3)표체적영향.방법 채용갑기특이성PCR법검측AS2O3작용전후MM세포주U266화CZ-1세포내SOCS-1기인적갑기화상태,응용단백면역인적법검측AS2O3처리전후세포내P-STAT3단백적표체변화,병채용류식세포기술검측AS2O3작용전후MM세포증식화조망적변화.결과 MM세포주내SOCS-1기인존재정도불동적갑기화상태,여대조조상비,AS2O3작용후MM세포내SOCS-1기인갑기화정도명현감약혹소실,P-STAT3단백적표체야명현감약,동시세포생장수억,조망비솔승고.AS2O3농도분별위0、0.5、1.0、2.0μmol/L시,U266세포주적총조망솔분별위0.06％、0.56％、48.96％、61.07％(x2=9.19,P＜0.05)；이CZ-1세포주적총조망솔분별위4.20％、40.30％、47.72％、68.49％(x2=8.96,P＜0.05).결론AS2O3가능통과유도MM세포내SOCS-1기인거갑기화작용,진일보억제세포증식신호Janus격매(JAK) -STAT통로적활화,종이유도MM세포적조망.
Objective To investigate the effects of arsenic trioxide (AS2O3)on SOCS-1 gene methylation and expression of P-STAT3 in multiple myeloma (MM) cells.Methods MM cell lines U266 and CZ-1 were used as in vitro models.Methylation status of SOCS-1 gene was detected by the methylation specific PCR (MSP)while P-STAT3 protein expression was determined by Western blotting assay before and after AS2O3 treatment.Meanwhile growth inhibition and apoptosis of MM cells were determined by flow cytometry.Results Hypermethylation of SOCS-1 gene was observed in each MM cell line compared with wide type.After exposure to AS2O3,it was shown that SOCS-1 gene was demethylated obviously,meanwhile the expression level of P-STAT3 protein and cell proliferation was inhibited significantly in each cell line.The apoptosis rate was increased.When U266 and CZ-1 were treated with AS2O3 of 0,0.5,1.0,2.0 μmol/L respectively,the total cell apoptosisis ratio of U266 was 0.06％,0.56％,48.96％,61.07％ (X2 =9.19,P ＜ 0.05); and the total cell apoptosisis ratio of CZ-1 was 4.2％,,40.3％,,47.72％,,68.49％ (X2 =8.96,P ＜0.05 ).Conclusion AS2O3 could inhibit JAK/STAT signal transduction pathway by inducing SOCS-1 gene demethylation in MM cells which might be related to cell apoptosis.