中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2012年
11期
896-901
,共6页
王翠翠%怀磊%张翠萍%贾玉娇%李其辉%陈一瑞%田征%唐克晶%邢海燕%王敏%王建祥
王翠翠%懷磊%張翠萍%賈玉嬌%李其輝%陳一瑞%田徵%唐剋晶%邢海燕%王敏%王建祥
왕취취%부뢰%장취평%가옥교%리기휘%진일서%전정%당극정%형해연%왕민%왕건상
白血病,急性%基因,PTEN%假基因
白血病,急性%基因,PTEN%假基因
백혈병,급성%기인,PTEN%가기인
Leukemia,acute%Gene,PTEN%Pseudogene
目的 探讨抑癌基因PTEN与其假基因PTENP1在急性白血病(AL)细胞中的表达情况及两者之间的相关性,并观察在白血病细胞系中PTENP1对PTEN的调控作用.方法 应用实时定量PCR方法检测138例初诊AL患者及15名正常对照者骨髓细胞中PTEN与PTENP1 mRNA的表达水平.构建PTENP1 3'UTR的慢病毒载体pCDH1-PTENP1 3'UTR-GFP,磷酸钙共沉淀法转染293T细胞,病毒原液感染HL-60细胞,流式细胞术分选获得混合克隆.实时定量PCR检测两组不同克隆PTENP1mRNA表达变化,Western blot检测两组不同克隆PTEN蛋白表达变化,MTT法检测细胞增殖抑制率.结果 与正常对照组相比,AL患者骨髓细胞PTEN及PTENPl mRNA表达水平均明显降低(P＜0.05).相关性分析显示,PTEN与PTENP1 mRNA表达水平呈正相关(P＜0.05).将PTENP1阳性急性髓系白血病(AML)组患者(108例)按NCCN 2011年AML治疗指南进行预后分组,各组之间PTEN及PTENP1表达水平无明显差异.pCDH1-PTENP1 3'UTR-GFP病毒原液感染的HL-60细胞,与对照组(pCDHl-GFP)相比,前者感染的HL-60细胞PTENPl mRNA水平明显升高,PTEN mRNA水平亦有升高,但PTEN蛋白表达和细胞增殖无明显变化.结论 AL患者骨髓细胞PTEN及PTENPl mRNA表达水平降低,两者的表达存在相关性.PTENP1可能在mRNA水平对PTEN的表达具有调控作用.
目的 探討抑癌基因PTEN與其假基因PTENP1在急性白血病(AL)細胞中的錶達情況及兩者之間的相關性,併觀察在白血病細胞繫中PTENP1對PTEN的調控作用.方法 應用實時定量PCR方法檢測138例初診AL患者及15名正常對照者骨髓細胞中PTEN與PTENP1 mRNA的錶達水平.構建PTENP1 3'UTR的慢病毒載體pCDH1-PTENP1 3'UTR-GFP,燐痠鈣共沉澱法轉染293T細胞,病毒原液感染HL-60細胞,流式細胞術分選穫得混閤剋隆.實時定量PCR檢測兩組不同剋隆PTENP1mRNA錶達變化,Western blot檢測兩組不同剋隆PTEN蛋白錶達變化,MTT法檢測細胞增殖抑製率.結果 與正常對照組相比,AL患者骨髓細胞PTEN及PTENPl mRNA錶達水平均明顯降低(P＜0.05).相關性分析顯示,PTEN與PTENP1 mRNA錶達水平呈正相關(P＜0.05).將PTENP1暘性急性髓繫白血病(AML)組患者(108例)按NCCN 2011年AML治療指南進行預後分組,各組之間PTEN及PTENP1錶達水平無明顯差異.pCDH1-PTENP1 3'UTR-GFP病毒原液感染的HL-60細胞,與對照組(pCDHl-GFP)相比,前者感染的HL-60細胞PTENPl mRNA水平明顯升高,PTEN mRNA水平亦有升高,但PTEN蛋白錶達和細胞增殖無明顯變化.結論 AL患者骨髓細胞PTEN及PTENPl mRNA錶達水平降低,兩者的錶達存在相關性.PTENP1可能在mRNA水平對PTEN的錶達具有調控作用.
목적 탐토억암기인PTEN여기가기인PTENP1재급성백혈병(AL)세포중적표체정황급량자지간적상관성,병관찰재백혈병세포계중PTENP1대PTEN적조공작용.방법 응용실시정량PCR방법검측138례초진AL환자급15명정상대조자골수세포중PTEN여PTENP1 mRNA적표체수평.구건PTENP1 3'UTR적만병독재체pCDH1-PTENP1 3'UTR-GFP,린산개공침정법전염293T세포,병독원액감염HL-60세포,류식세포술분선획득혼합극륭.실시정량PCR검측량조불동극륭PTENP1mRNA표체변화,Western blot검측량조불동극륭PTEN단백표체변화,MTT법검측세포증식억제솔.결과 여정상대조조상비,AL환자골수세포PTEN급PTENPl mRNA표체수평균명현강저(P＜0.05).상관성분석현시,PTEN여PTENP1 mRNA표체수평정정상관(P＜0.05).장PTENP1양성급성수계백혈병(AML)조환자(108례)안NCCN 2011년AML치료지남진행예후분조,각조지간PTEN급PTENP1표체수평무명현차이.pCDH1-PTENP1 3'UTR-GFP병독원액감염적HL-60세포,여대조조(pCDHl-GFP)상비,전자감염적HL-60세포PTENPl mRNA수평명현승고,PTEN mRNA수평역유승고,단PTEN단백표체화세포증식무명현변화.결론 AL환자골수세포PTEN급PTENPl mRNA표체수평강저,량자적표체존재상관성.PTENP1가능재mRNA수평대PTEN적표체구유조공작용.
Objective To investigate the expression of PTEN(phosphatase and tension homology deletion on chromosome 10,PTEN) and its pseudogene PTENP1 in acute leukemia (AL) and correlation between them,and to explore the role of PTENP1 on the PTEN expression in AL cells.Methods PTEN and PTENP1 mRNA expression were evaluated in bone marrow (BM) samples from 138 newly diagnosed AL patients and 15 healthy controls by quantitative real-time RT-PCR(qRT-PCR).pCDH1-PTENP1 3'UTR-GFP lentivirus vectors were constructed.293T cells were transfected by calcium phosphate precipitation to produce retrovirus.HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP and pCDH1-PTENP1 3'UTR-GFP respectively.The flow cell sorter was used to sort the HL-60 with GFP positively expressed.The mRNA expression of PTEN and PTENP1 was detected by qRT-PCR,the expression of PTEN protein by western blot,and the impact of PTENP13'UTR on the proliferation of HL-60 cells by MTT assay.Results AML patients showed significantly lower PTEN and PTENP1 mRNA expression in BM compared to healthy controls.Correlation analysis showed that the expression of PTEN and PTENP1 mRNA were positively correlated(P ＜ 0.05).The 108 cases of PTENP1 + AML were classified according to the prognostic classification of 2011 NCCN Clinical Practice Guidelines in AML,there was no difference among different subgroups.HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP (control group) and pCDH1-PTENP1 3'UTR-GFP respectively.Compared with the control group,PTENP1 mRNA level of HL-60 infected with the retroviral vectors expressing pCDH1-PTENP1 3' UTR-GFP increased significantly,and PTEN mRNA level also increased.While the PTEN protein level and the cell growth rate of the PTENP1 3'UTR group didn' t change significantly.Conclusion PTEN and PTENP1 mRNA expression level of BM cells from AL patients is significantly lower.There is a positive correlation between expression of PTEN and PTENP1 mRNA.PTENP1 may regulate the expression of PTEN in mRNA level.
