中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2015年
11期
947-950
,共4页
张贵丽%董菲%栾材富%张霞%邵会媛%刘杰%孙成铭
張貴麗%董菲%欒材富%張霞%邵會媛%劉傑%孫成銘
장귀려%동비%란재부%장하%소회원%류걸%손성명
白血病,髓系,慢性,BCR-ABL阳性%CCAAT增强子结合蛋白α%K562细胞%细胞增殖
白血病,髓繫,慢性,BCR-ABL暘性%CCAAT增彊子結閤蛋白α%K562細胞%細胞增殖
백혈병,수계,만성,BCR-ABL양성%CCAAT증강자결합단백α%K562세포%세포증식
Leukemia,myelogenous,chronic,BCR-ABL positive%CCAAT-enhancer-binding protein-alpha%K562 cell%Cell growth
目的 研究转录因子CCAAT/增强子结合蛋白α(C/EBPα)在慢性髓性白血病(CML)患者中的表达情况及其作用机制.方法 收集50例CML患者(慢性期33例,加速期7例,急变期10例)骨髓标本和20名正常对照者外周血标本,用RT-PCR方法检测C/EBPα基因mRNA的表达情况及其与bcr-abl融合基因的相关性;观察伊马替尼对K562细胞C/EBPα基因mRNA表达的影响;采用慢病毒转染的方法将重组pLVX-C/EBPα-3FLAG-Puro及空载体pLVX-EGFP-3FLAG-Puro慢病毒质粒转入K562细胞,构建稳定表达C/EBPα的K562细胞株;用CCK-8试剂盒检测细胞增殖;RT-PCR方法检测Foxo3a、Bim基因的表达情况.结果 与正常对照者相比,CML患者C/EBPα mRNA表达显著下降(P＜0.01),并与bcr-abl融合基因表达呈负相关(r=-0.505,P＜0.01);RT-PCR和Western blot检测结果显示成功构建稳定表达C/EBPα的K562细胞株;K562-C/EBPα细胞增殖水平明显低于未处理K562细胞及K562-空载体细胞;K562-C/EBPα细胞Foxo3a、Bim基因mRNA相对表达量分别为1.06±0.06、0.53±0.07,均高于K562-空载体和K562细胞(P＜0.01,P＜0.05).结论 CML患者转录因子C/EBPα表达水平下调,过表达C/EBPα对K562细胞增殖有抑制作用.
目的 研究轉錄因子CCAAT/增彊子結閤蛋白α(C/EBPα)在慢性髓性白血病(CML)患者中的錶達情況及其作用機製.方法 收集50例CML患者(慢性期33例,加速期7例,急變期10例)骨髓標本和20名正常對照者外週血標本,用RT-PCR方法檢測C/EBPα基因mRNA的錶達情況及其與bcr-abl融閤基因的相關性;觀察伊馬替尼對K562細胞C/EBPα基因mRNA錶達的影響;採用慢病毒轉染的方法將重組pLVX-C/EBPα-3FLAG-Puro及空載體pLVX-EGFP-3FLAG-Puro慢病毒質粒轉入K562細胞,構建穩定錶達C/EBPα的K562細胞株;用CCK-8試劑盒檢測細胞增殖;RT-PCR方法檢測Foxo3a、Bim基因的錶達情況.結果 與正常對照者相比,CML患者C/EBPα mRNA錶達顯著下降(P＜0.01),併與bcr-abl融閤基因錶達呈負相關(r=-0.505,P＜0.01);RT-PCR和Western blot檢測結果顯示成功構建穩定錶達C/EBPα的K562細胞株;K562-C/EBPα細胞增殖水平明顯低于未處理K562細胞及K562-空載體細胞;K562-C/EBPα細胞Foxo3a、Bim基因mRNA相對錶達量分彆為1.06±0.06、0.53±0.07,均高于K562-空載體和K562細胞(P＜0.01,P＜0.05).結論 CML患者轉錄因子C/EBPα錶達水平下調,過錶達C/EBPα對K562細胞增殖有抑製作用.
목적 연구전록인자CCAAT/증강자결합단백α(C/EBPα)재만성수성백혈병(CML)환자중적표체정황급기작용궤제.방법 수집50례CML환자(만성기33례,가속기7례,급변기10례)골수표본화20명정상대조자외주혈표본,용RT-PCR방법검측C/EBPα기인mRNA적표체정황급기여bcr-abl융합기인적상관성;관찰이마체니대K562세포C/EBPα기인mRNA표체적영향;채용만병독전염적방법장중조pLVX-C/EBPα-3FLAG-Puro급공재체pLVX-EGFP-3FLAG-Puro만병독질립전입K562세포,구건은정표체C/EBPα적K562세포주;용CCK-8시제합검측세포증식;RT-PCR방법검측Foxo3a、Bim기인적표체정황.결과 여정상대조자상비,CML환자C/EBPα mRNA표체현저하강(P＜0.01),병여bcr-abl융합기인표체정부상관(r=-0.505,P＜0.01);RT-PCR화Western blot검측결과현시성공구건은정표체C/EBPα적K562세포주;K562-C/EBPα세포증식수평명현저우미처리K562세포급K562-공재체세포;K562-C/EBPα세포Foxo3a、Bim기인mRNA상대표체량분별위1.06±0.06、0.53±0.07,균고우K562-공재체화K562세포(P＜0.01,P＜0.05).결론 CML환자전록인자C/EBPα표체수평하조,과표체C/EBPα대K562세포증식유억제작용.
Objective To investigate the expression and the possible mechanism of the transcription factor C/EBPα in chronic myeloid leukemia (CML).Methods Bone marrow samples from 50 CML patients (including 33 patients in chronic phase,7 in accelerated phase and 10 in blast crisis) and peripheral blood specimens of 20 healthy donors were collected.The expression of C/EBPα gene and the effect of Imatinib on its expression was detected by RT-PCR.C/EBPα gene was inserted into lentivirus expression vector pLVX-EGFP-3FLAG-Puro by recombinant DNA technology to construct C/EBPα stable expression in K562 cells.Cell proliferation was assayed by CCK-8.The expressions of Foxo3a and Bim genes were detected by RT-PCR.Results The level of C/EBPαt expression was significantly declined in CML patients compared with that of normal control group (P＜0.01) and had negative correlation with bcr-abl expression (Spearman r=-0.505,P＜0.01).The stable K562-C/EBPα cell line was successfully established and confirmed by RT-PCR and Western blot.Cell proliferation ability was lower in the K562-C/EBPα group than that in the non-transfection and mock-vehicle groups.The expressions of Foxo3a and Bim genes were 1.06 ± 0.06 and 0.53 ± 0.07,respectively,which was higher than that of non-transfection and mock-vehicle groups (P＜0.01,P＜0.05).Conclusion C/EBPα expression was decreased in CML patients,overexpression of C/EBPα could inhibit K562 cell growth.
