中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
5期
317-321
,共5页
王晔恺%周吉航%周世权%方国安%李翊卫%邱雷%杨序春%刘晓光
王曄愷%週吉航%週世權%方國安%李翊衛%邱雷%楊序春%劉曉光
왕엽개%주길항%주세권%방국안%리익위%구뢰%양서춘%류효광
基因,hPer3%白血病,非淋巴细胞,急性%甲基化聚合酶链反应%地西他滨
基因,hPer3%白血病,非淋巴細胞,急性%甲基化聚閤酶鏈反應%地西他濱
기인,hPer3%백혈병,비림파세포,급성%갑기화취합매련반응%지서타빈
Gene,hPer3%Leukemia,nonlymphocytic,acute%Methylation specific-PCR%Decitabine
目的 探讨急性髓系白血病(AML)患者hPer3基因启动子甲基化检测的临床意义,并观察地西他滨(DCA)诱导hPer3基冈表达对AML细胞系HL-60和U937增殖的影响.方法 应用甲基化聚合酶链反应(MS-PCR)和实时荧光定量PCR(RT-PCR)检测206例AML患者骨髓细胞hPer3基因启动子甲基化状态和mRNA表达,以40例缺铁性贫血患者骨髓细胞作为对照.用不同浓度的DCA处理HL-60和U937细胞48和72 h,检测其hPer3基因启动子甲基化状态及其mRNA表达,用MTT法检测细胞增殖抑制率,Annexin V-FITC/PI标记法检测细胞凋亡,用流式细胞术检测CD14、CD11b抗原表达.结果 AML患者初发组、部分缓解组、完全缓解组、复发组中,hPer3基因启动子甲基化阳性率分别为93.65%(63例中59例)、54.39%(57例中31例)、24.66%(73例中18例)、61.54%(13例中8例),对照组无一例甲基化;hPer3 mRNA表达分别为0.19±0.08、6.28±2.11、52.76±14.17、8.18±4.36,明显低于对照组(75.03±18.16),除部分缓解组与复发组相比差异无统计学意义(P>0.05)外,其余组别两两比较差异均有统计学意义(P<0.01).DCA处理HL-60和U937细胞后,hPer3基因启动子甲基化水平降低,mRNA表达升高,细胞出现凋亡,CD14和CD11b表达升高,并呈剂量和时间依赖性.结论 AML患者骨髓细胞hPer3基因启动子甲基化状态和mRNA表达水平可能作为AML患者病情缓解程度的一个评价指标.体外应用DCA有助于诱导hPer3基因表达和促AML细胞凋亡.
目的 探討急性髓繫白血病(AML)患者hPer3基因啟動子甲基化檢測的臨床意義,併觀察地西他濱(DCA)誘導hPer3基岡錶達對AML細胞繫HL-60和U937增殖的影響.方法 應用甲基化聚閤酶鏈反應(MS-PCR)和實時熒光定量PCR(RT-PCR)檢測206例AML患者骨髓細胞hPer3基因啟動子甲基化狀態和mRNA錶達,以40例缺鐵性貧血患者骨髓細胞作為對照.用不同濃度的DCA處理HL-60和U937細胞48和72 h,檢測其hPer3基因啟動子甲基化狀態及其mRNA錶達,用MTT法檢測細胞增殖抑製率,Annexin V-FITC/PI標記法檢測細胞凋亡,用流式細胞術檢測CD14、CD11b抗原錶達.結果 AML患者初髮組、部分緩解組、完全緩解組、複髮組中,hPer3基因啟動子甲基化暘性率分彆為93.65%(63例中59例)、54.39%(57例中31例)、24.66%(73例中18例)、61.54%(13例中8例),對照組無一例甲基化;hPer3 mRNA錶達分彆為0.19±0.08、6.28±2.11、52.76±14.17、8.18±4.36,明顯低于對照組(75.03±18.16),除部分緩解組與複髮組相比差異無統計學意義(P>0.05)外,其餘組彆兩兩比較差異均有統計學意義(P<0.01).DCA處理HL-60和U937細胞後,hPer3基因啟動子甲基化水平降低,mRNA錶達升高,細胞齣現凋亡,CD14和CD11b錶達升高,併呈劑量和時間依賴性.結論 AML患者骨髓細胞hPer3基因啟動子甲基化狀態和mRNA錶達水平可能作為AML患者病情緩解程度的一箇評價指標.體外應用DCA有助于誘導hPer3基因錶達和促AML細胞凋亡.
목적 탐토급성수계백혈병(AML)환자hPer3기인계동자갑기화검측적림상의의,병관찰지서타빈(DCA)유도hPer3기강표체대AML세포계HL-60화U937증식적영향.방법 응용갑기화취합매련반응(MS-PCR)화실시형광정량PCR(RT-PCR)검측206례AML환자골수세포hPer3기인계동자갑기화상태화mRNA표체,이40례결철성빈혈환자골수세포작위대조.용불동농도적DCA처리HL-60화U937세포48화72 h,검측기hPer3기인계동자갑기화상태급기mRNA표체,용MTT법검측세포증식억제솔,Annexin V-FITC/PI표기법검측세포조망,용류식세포술검측CD14、CD11b항원표체.결과 AML환자초발조、부분완해조、완전완해조、복발조중,hPer3기인계동자갑기화양성솔분별위93.65%(63례중59례)、54.39%(57례중31례)、24.66%(73례중18례)、61.54%(13례중8례),대조조무일례갑기화;hPer3 mRNA표체분별위0.19±0.08、6.28±2.11、52.76±14.17、8.18±4.36,명현저우대조조(75.03±18.16),제부분완해조여복발조상비차이무통계학의의(P>0.05)외,기여조별량량비교차이균유통계학의의(P<0.01).DCA처리HL-60화U937세포후,hPer3기인계동자갑기화수평강저,mRNA표체승고,세포출현조망,CD14화CD11b표체승고,병정제량화시간의뢰성.결론 AML환자골수세포hPer3기인계동자갑기화상태화mRNA표체수평가능작위AML환자병정완해정도적일개평개지표.체외응용DCA유조우유도hPer3기인표체화촉AML세포조망.
Objective To investigate the clinical significance of promoter methylation status of hPer3 gene in acute myeloid leukemia( AML) patients and the in vitro effect of decitabine( DCA) on AML cell lines HL-60 and U937. Methods The promoter methylation status of hPer3 gene and mRNA expression levels in bone marrow of 206 AML and 40 iron deficiency anemia( IDA) patients ( as control) were detected by methylation specific PCR(MS-PCR) and real-time PCR(RT-PCR). The HL-60 and U937 cell lines were treated with different concertrations of DCA for 48 and 72 h. The inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium(MTT) ;the early apoptosis rates by staining with Annexin V and PI; the CD14 and CD11b expressions by flow cytometry( FCM) ; the promoter methylation status of hPer3 gene by MS-PCR;and the hPer3 mRNA expressions levels by RT-PCR. Results The promoter methylation rates of hPer3 in newly diagnosed( ND) group, partial remission ( PR) group, complete remission ( CR ) group, relapse (R)group and control group were 93.65% (59/63) ,54. 39% (31/57) ,24. 66% (18/73) ,61. 54% (8/13) and 0% (0/40), and the hPer3 mRNA expression levels were 0. 19 ±0.08,6.28 ±2. 11,52.76 ± 14.17,8. 18 ±4.36,75.03 ±18. 16, respeatively. There was a significant statistic difference between any two group (P <0.01) excepting for between PR and R group(P>0.05). After DCA treatment,the promoter hypermethylation status of hPer3 was reduced and the mRNA and CD14,CD11b expression levels were up regulated in a dose dependent manner with an induction of cell apoptosis. Conclusions Promoter methylation status and mRNA expression of hPer3 gene may be indicators for evaluating AML. DCA can induce the expression of hPer3 gene and cells apoptosis in AML.
