CAJ | 학술논문

目的探讨急性白血病患者组蛋白乙酰化修饰规律,并探索组蛋白乙酰化对错配修复基因hMSH2和hMLH1差异表达的调控作用.方法用反转录-聚合酶链反应(RT-PCR)方法检测56例急性白血病患者和30名健康志愿者单个核细胞(MNC)的错配修复基因hMSH2和hMLH1 mRNA的表达,用Western blot法检测组蛋白H3、H4、去乙酰化酶(HDAC1)、hMSH2和hMLH1基因的蛋白表达情况.用组蛋白去乙酰转移酶抑制剂(TSA)诱导30例白血病患者MNC乙酰化,并检测处理后MNC的组蛋白H3、H4、HDAC1、hMSH2和hMLH1的表达状态变化.结果急性白血病组的hMSH2和hMLH1、组蛋白H3、H4的蛋白表达量分别为0.4610±0.1211、0.4013±0.1143、0.4103±0.1241和0.4251±0.1081,均明显低于健康志愿者组的蛋白表达量(0.9461±0.1841、0.9960±0.2021、0.8971±0.1194、0.9513±0.1953),差异均有统计学意义(t值分别为3.341、3.935、2.843、3.575,P＜0.05);而急性白血病患者组的HDAC1表达(0.8841±0.2018)高于健康志愿者组的表达量(0.5142±0.1340),差异有统计学意义(t=2.634,P＜0.05);TSA作用于白血病单个核细胞后,组蛋白H3、H4、hMSH2和hMLH1的表达上调,分别比阴性对照组表达上调2.9倍、3.4倍、1.5倍和1.6倍,而HDAC1的蛋白表达出现明显的抑制,表达下调为阴性对照组的40%.结论急性白血病患者的组蛋白乙酰化呈低表达现象,组蛋白乙酰化在急性白血病患者中对错配修复基因差异表达具有调控作用.
목적 탐토급성백혈병환자조단백을선화수식규률,병탐색조단백을선화대착배수복기인hMSH2화hMLH1차이표체적조공작용.방법 용반전록-취합매련반응(RT-PCR)방법검측56례급성백혈병환자화30명건강지원자단개핵세포(MNC)적착배수복기인hMSH2화hMLH1 mRNA적표체,용Western blot법검측조단백H3、H4、거을선화매(HDAC1)、hMSH2화hMLH1기인적단백표체정황.용조단백거을선전이매억제제(TSA)유도30례백혈병환자MNC을선화,병검측처리후MNC적조단백H3、H4、HDAC1、hMSH2화hMLH1적표체상태변화.결과 급성백혈병조적hMSH2화hMLH1、조단백H3、H4적단백표체량분별위0.4610±0.1211、0.4013±0.1143、0.4103±0.1241화0.4251±0.1081,균명현저우건강지원자조적단백표체량(0.9461±0.1841、0.9960±0.2021、0.8971±0.1194、0.9513±0.1953),차이균유통계학의의(t치분별위3.341、3.935、2.843、3.575,P＜0.05);이급성백혈병환자조적HDAC1표체(0.8841±0.2018)고우건강지원자조적표체량(0.5142±0.1340),차이유통계학의의(t=2.634,P＜0.05);TSA작용우백혈병단개핵세포후,조단백H3、H4、hMSH2화hMLH1적표체상조,분별비음성대조조표체상조2.9배、3.4배、1.5배화1.6배,이HDAC1적단백표체출현명현적억제,표체하조위음성대조조적40%.결론 급성백혈병환자적조단백을선화정저표체현상,조단백을선화재급성백혈병환자중대착배수복기인차이표체구유조공작용.
Objective To explore the status of histone acetylation modification and their regulatory effect to hMSH2 gene and hMLH1 gene expression in acute leukemia. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of hMSH2 and hMLH1 mRNA, and Western blot was used to measure the expression of histone H3, H4, HDACi, hMSH2 and hMLH1 protein in mononuclear cells of 56 acute leukemia patients and 30 healthy volunteers. The mononuclear cells of 30 acute leukemia patients were treated with histone deacetylase inhibitors trichostatin A (TSA), and measured the expression difference of histone H3, H4, HDAC1, hMSH2 and hMLH1 in the mononuclear cells treated with TSA. Results The protein expression levels of hMSH2, hMLH1, histone H3 and histone H4 in those mononuclear cells of acute leukemia patients were 0.4610±0.1211, 0.4013±0.1143, 0.4103±0.1241 and 0.4251±0.1081, respectively, which were significantly decreased comparing with those of healthy volunteers (0.9461±0. 1841, 0.996±0.2021, 0.8971±0. 1194 and 0.9513±0.1953) (t = 3.341, 3.935, 2.843 and 3.575,respectinely, P ＜0.05). The protein expression levels of HDAC1 (0.8841±0.2018) of acute leukemia patients was significantly increased comparing with those of healthy volunteers (0.5142±0.1340) (t= 2.634, P ＜0.05).After treatment with TSA for 48 hours, the protein expression of hMSH2 was increased nearly 1.5-fold, hMLH1 about 1.6-fold, H3 about 2.9-fold and H4 about 3.4-fold comparing with the negative control groups (P ＜0.05),while the protein expression of HDAC1 were decreased comparing with the negative control groups by 40 %.Conclusion There was an low expression phenomenon of histone acetylation in acute leukemia, and histone acetylation played an important role in regulation of the mismatch repair gene expression in acute leukemia.