CAJ | 학술논문

目的探讨B细胞淋巴瘤中LITAF基因启动子区CpG岛异常甲基化情况,研究去甲基化药物对人B细胞淋巴瘤细胞系Raji、Pfeiffer和Daudi细胞中LITAF基因的转录调控作用,寻找肿瘤治疗新靶点.方法收集存档石蜡包埋组织标本105例[弥漫性大B细胞淋巴瘤(DLBCL) 54例,小淋巴细胞性淋巴瘤(SLL)15例,黏膜相关淋巴组织结外边缘区淋巴瘤(MALT)8例,滤泡性淋巴瘤(FL)6例,其他22例],5例良性反应性增生淋巴组织作为正常对照.甲基化特异性PCR(MSP)检测LITAF基因启动子区甲基化状态.运用MSP、Western blot、免疫组织化学方法检测5-氮杂脱氧胞苷处理前后Raji、Pfeiffer和Daudi细胞系的甲基化状态及蛋白表达情况.四甲基偶氮唑盐(MTT)法检测经去甲基化药物处理后细胞生长抑制情况.结果 B细胞淋巴瘤中89.5％(94/105)的病例存在LITAF基因启动子区高甲基化,其中3.8％ (4/105)的病例发生完全高甲基化,22.9％(24/105)的病例以部分非甲基化为主,对照组5例良性淋巴结增生标本中LITAF基因启动子区均未检测到高甲基化改变.经去甲基化处理后,Raji、Daudi、Pfeiffer细胞LITAF基因甲基化程度明显受抑制,基因表达得以恢复和增加.结论在B细胞淋巴瘤患者中普遍存在LITAF基因启动子区CpG岛高甲基化,由其所致的LITAF基因表达沉默或减少可能是B细胞淋巴瘤发生的一个重要因素,LITAF基因的高甲基化可望作为一个良好的候选分子诊断标记及可能的治疗靶点.
목적 탐토B세포림파류중LITAF기인계동자구CpG도이상갑기화정황,연구거갑기화약물대인B세포림파류세포계Raji、Pfeiffer화Daudi세포중LITAF기인적전록조공작용,심조종류치료신파점.방법 수집존당석사포매조직표본105례[미만성대B세포림파류(DLBCL) 54례,소림파세포성림파류(SLL)15례,점막상관림파조직결외변연구림파류(MALT)8례,려포성림파류(FL)6례,기타22례],5례량성반응성증생림파조직작위정상대조.갑기화특이성PCR(MSP)검측LITAF기인계동자구갑기화상태.운용MSP、Western blot、면역조직화학방법검측5-담잡탈양포감처리전후Raji、Pfeiffer화Daudi세포계적갑기화상태급단백표체정황.사갑기우담서염(MTT)법검측경거갑기화약물처리후세포생장억제정황.결과 B세포림파류중89.5％(94/105)적병례존재LITAF기인계동자구고갑기화,기중3.8％ (4/105)적병례발생완전고갑기화,22.9％(24/105)적병례이부분비갑기화위주,대조조5례량성림파결증생표본중LITAF기인계동자구균미검측도고갑기화개변.경거갑기화처리후,Raji、Daudi、Pfeiffer세포LITAF기인갑기화정도명현수억제,기인표체득이회복화증가.결론 재B세포림파류환자중보편존재LITAF기인계동자구CpG도고갑기화,유기소치적LITAF기인표체침묵혹감소가능시B세포림파류발생적일개중요인소,LITAF기인적고갑기화가망작위일개량호적후선분자진단표기급가능적치료파점.
Objective To investigate promoter methylation status of LITAF gene in B-cell lymphoma and to explore transcription regulation of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on LITAF gene.Methods One hundred and five paraffin specimens including 54 cases of diffuse large B cell lymphoma (DLBCL),15 small lymphocytic lymphoma (SLL),8 mucosa-associated lymphoid tissue lymphoma (MALT) and 6 follicular lymphoma (FL) were included.Five reactive lymphoid hyperplasia samples were collected as control.Methylation status of CpG island in LITAF gene in the specimens and in Raji,Pfeiffer and Daudi cell lines were detected by methylation-specific PCR (MSP).LITAF expression in Raji,Pfeiffer and Daudi cell lines with or without 5-Aza-CdR treatment was detected by Western blot and immunohistochemistry.The inhibitory ratio in the three cell lines was measured by MTT assay.Results The frequency of LITAF gene methylation in B-cell lymphoma was 89.5％ (94/105).Among them,3.8％ (4/105) showed complete hypermethylation.In control group,however,there was no methylation in CpG island of LITAF gene promoter.The expression of LITAF was recovered or increased along with the cell growth inhibition when the cells exposed to demethylating reagent.Conclusions LITAF gene silencing with aberrant CpG methylation is probably one of the critical events to the oncogenesis of B-cell lymphoma,which may have important implications as a candidate marker for diagnosis and target gene therapy.