CAJ | 학술논문

目的检测骨髓嗜病毒整合位点1(MEIS1)基因在同源异形盒11(HOX11)+急性T淋巴细胞白血病(T-ALL)细胞株和T-ALL患者骨髓细胞中的甲基化状态和表达水平,探讨其表达水平与其启动子甲基化的关系.方法收集2009年1月至2011年12月北京大学人民医院北京大学血液病研究所38例T-ALL患者治疗前及其中29例治疗后完全缓解的T-ALL患者和8名健康供者的骨髓样本,及3株T-ALL细胞株(sil-ALL、DND41和RPMI);采用反转录(RT) PCR检测MEIS1、HOX11mRNA表达水平,采用重硫酸盐测序法、甲基化DNA免疫沉淀技术和启动子寡核苷酸微阵列芯片分析MEIS1基因启动子区CpG岛甲基化状态和基因功能.结果根据HOX11表达的高低,将38例T-ALL患者分为HOX11高表达(HOX11+)组(n=14)和HOX11低表达(HOX11-)组(n=24).HOX11+组患者骨髓中出现MEIS1基因启动子区域的甲基化.HOX11+组患者MEIS1基因启动子区域甲基化比率明显高于HOX11-组患者(76.8％比29.5％,P＜0.01).MEIS1 mRNA在治疗后完全缓解患者和健康供者骨髓中的水平低于患者治疗前骨髓中的水平(均P ＜0.05);MEIS1 mRNA在HOX11+ T-ALL细胞株(sil-ALL、DND41)中的表达明显低于HOX 11-T-ALL细胞株(RPMI) (0.392±0.226、0.378 ±0.317比1.059±0.533,均P＜0.05),在HOX11+组患者中的表达明显低于HOX11-组患者(0.462±0.281比0.916±0.437,P＜0.01);且MEIS1 mRNA的相对表达程度与HOX11mRNA的相对表达程度呈负相关(rs=-0.416,P＜0.01).MEIS1基因在T-ALL中的表达水平与其甲基化呈负相关(rs=-0.383,P＜0.01).结论 MEIS1基因启动子甲基化是导致其在T-ALL中表达下调或基因沉默的原因,HOX11可能负调控MEIS1基因的表达.
목적 검측골수기병독정합위점1(MEIS1)기인재동원이형합11(HOX11)+급성T림파세포백혈병(T-ALL)세포주화T-ALL환자골수세포중적갑기화상태화표체수평,탐토기표체수평여기계동자갑기화적관계.방법 수집2009년1월지2011년12월북경대학인민의원북경대학혈액병연구소38례T-ALL환자치료전급기중29례치료후완전완해적T-ALL환자화8명건강공자적골수양본,급3주T-ALL세포주(sil-ALL、DND41화RPMI);채용반전록(RT) PCR검측MEIS1、HOX11mRNA표체수평,채용중류산염측서법、갑기화DNA면역침정기술화계동자과핵감산미진렬심편분석MEIS1기인계동자구CpG도갑기화상태화기인공능.결과 근거HOX11표체적고저,장38례T-ALL환자분위HOX11고표체(HOX11+)조(n=14)화HOX11저표체(HOX11-)조(n=24).HOX11+조환자골수중출현MEIS1기인계동자구역적갑기화.HOX11+조환자MEIS1기인계동자구역갑기화비솔명현고우HOX11-조환자(76.8％비29.5％,P＜0.01).MEIS1 mRNA재치료후완전완해환자화건강공자골수중적수평저우환자치료전골수중적수평(균P ＜0.05);MEIS1 mRNA재HOX11+ T-ALL세포주(sil-ALL、DND41)중적표체명현저우HOX 11-T-ALL세포주(RPMI) (0.392±0.226、0.378 ±0.317비1.059±0.533,균P＜0.05),재HOX11+조환자중적표체명현저우HOX11-조환자(0.462±0.281비0.916±0.437,P＜0.01);차MEIS1 mRNA적상대표체정도여HOX11mRNA적상대표체정도정부상관(rs=-0.416,P＜0.01).MEIS1기인재T-ALL중적표체수평여기갑기화정부상관(rs=-0.383,P＜0.01).결론 MEIS1기인계동자갑기화시도치기재T-ALL중표체하조혹기인침묵적원인,HOX11가능부조공MEIS1기인적표체.
Objective To detect the status of methylation in promoter region of MEIS1 gene and its expression in cell lines and patients with HOX11 + T-cell acute lymphoblastic leukemia (T-ALL) and explore the relationship between the expression level of MEIS1 gene and methylation of CpG island in promoter region.Methods The methylation pattern in MEIS1 gene was detected with bisulfite sequencing PCR,DNA methylation immunoprecipitation and promoter oligonucleotide microarray.And the expression level of MEIS1 mRNA was detected with reverse transcription PCR in 3 T-ALL cell lines:sil-ALL,DND41 and RPMI as well as in 75 clinical bone marrow samples including 38 de novo T-ALL patients,29 complete remission T-ALL patients and 8 normal samples from January 2009 to December 2011.Results The expressions of HOX11 mRNA in the patients were classified into high expression(HOX11 +)groups(n =14) and low expression(HOX11-)groups(n =24).There was hypermethylation of CpG island in promoter region of MEIS1 gene in patients with HOX11 + T-ALL.The methylation rate of promoter CpG islands of MEIS1 gene in HOX11 + T-ALL (76.8％) was significantly higher than that of HOX11-T-ALL (29.5％) (P ＜0.01).The expression MEIS1 in complete remission T-ALL patients and normal samples was significantly lower than that in de novo T-ALL patients (both P ＜ 0.05).The expression MEIS1 in HOX11 + cell lines was significantly lower than that in HOX11-cell lines (0.392 ±0.226,0.378 ±0.317 vs 1.059 ±0.533,both P ＜ 0.05) and the expression level of MEIS1 in HOX11 + T-ALL patients (0.462 ± 0.281) significantly lower than that in HOX11-T-ALL patients (0.916 ± 0.437) (P ＜ 0.01).The relationship between MEIS1 and HOX11 mRNA expression level had a negative correlation(rs =-0.416,P ＜ 0.01).The expression level of MEIS1 gene was negatively correlated with the methylation of CpG island in promoter region in T-ALL (rs =-0.383,P ＜ 0.01).Conclusions The expression of MEIS1 becomes downregulated or even lost by promoter methylation in T-ALL.HOX11 maybe a negative regulator of MEIS1 gene.