中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2012年
7期
546-551
,共6页
吴勇%李先芳%杨景辉%廖晓莹%陈元仲
吳勇%李先芳%楊景輝%廖曉瑩%陳元仲
오용%리선방%양경휘%료효형%진원중
细胞系,NB4%白血病,早幼粒细胞,急性%细胞分化%微小RNA
細胞繫,NB4%白血病,早幼粒細胞,急性%細胞分化%微小RNA
세포계,NB4%백혈병,조유립세포,급성%세포분화%미소RNA
Cell line,NB4%Leukemia,promyelocytic,acute%Cell differentiation%microRNA
目的 研究急性早幼粒细胞白血病(APL)细胞分化前后微小RNA(miRNA,miR)表达变化.方法 采用全反式维甲酸(ATRA)和三氧化二砷(As2O3)诱导APL细胞系NB4 细胞分化,瑞氏-姬姆萨染色观察细胞形态,流式细胞术检测细胞表面标志CD11b的表达,用实时定量RT-PCR检测miRNA表达谱miR-15b、miR-16、miR-34a、miR-107、miR-124a、miR-146、miR-155、miR-181a、miR-223、miR-342、let7c等miRNA的表达水平,用2-△△Ct法计算miRNA相对表达水平.收集15例APL初诊和15例APL缓解期患者骨髓单个核细胞(MNC),用RT-PCR检测MNC miRNA 的表达水平,用2-△Ct法计算miRNA表达水平.结果 ATRA作用NB4细胞96 h,miR-15b、miR-16、miR-107、miR-223、miR-342表达水平均显著上调,分别为对照组的3.40、4.22、5.41、20.03和5.29倍,As2O3作用NB4细胞96 h,miR-15b、miR-16、miR-107、miR-223、miR-342表达水平也显著上调,分别为对照组的3.62、2.49、2.58、4.27和1.94倍,除miR-15b外,ATRA处理组miR-16、miR-107、miR-223和miR-342表达水平上调程度均高于As2O3处理组,其中尤以miR-223为甚.ATRA和As2O3治疗后缓解期APL患者miR-15b、miR-16、miR-107、miR-181a、miR-223和miR-342表达水平(2-△Ct值)分别为0.4137、0.6367、0.1260、0.0522、0.6611和0.0280,而APL初诊患者miR-15b、miR-16、miR-107、miR-181a、miR-223、miR-342表达水平分别为0.0751、0.2022、0.0425、0.3064、0.1733和0.0090,APL缓解期miR-15b、miR-16、miR-107、miR-223和miR-342表达水平高于APL初诊者(P值均<0.05),而APL缓解期miR-181a表达水平低于APL初诊者(P<0.05).结论 特定miRNA参与APL细胞分化过程.
目的 研究急性早幼粒細胞白血病(APL)細胞分化前後微小RNA(miRNA,miR)錶達變化.方法 採用全反式維甲痠(ATRA)和三氧化二砷(As2O3)誘導APL細胞繫NB4 細胞分化,瑞氏-姬姆薩染色觀察細胞形態,流式細胞術檢測細胞錶麵標誌CD11b的錶達,用實時定量RT-PCR檢測miRNA錶達譜miR-15b、miR-16、miR-34a、miR-107、miR-124a、miR-146、miR-155、miR-181a、miR-223、miR-342、let7c等miRNA的錶達水平,用2-△△Ct法計算miRNA相對錶達水平.收集15例APL初診和15例APL緩解期患者骨髓單箇覈細胞(MNC),用RT-PCR檢測MNC miRNA 的錶達水平,用2-△Ct法計算miRNA錶達水平.結果 ATRA作用NB4細胞96 h,miR-15b、miR-16、miR-107、miR-223、miR-342錶達水平均顯著上調,分彆為對照組的3.40、4.22、5.41、20.03和5.29倍,As2O3作用NB4細胞96 h,miR-15b、miR-16、miR-107、miR-223、miR-342錶達水平也顯著上調,分彆為對照組的3.62、2.49、2.58、4.27和1.94倍,除miR-15b外,ATRA處理組miR-16、miR-107、miR-223和miR-342錶達水平上調程度均高于As2O3處理組,其中尤以miR-223為甚.ATRA和As2O3治療後緩解期APL患者miR-15b、miR-16、miR-107、miR-181a、miR-223和miR-342錶達水平(2-△Ct值)分彆為0.4137、0.6367、0.1260、0.0522、0.6611和0.0280,而APL初診患者miR-15b、miR-16、miR-107、miR-181a、miR-223、miR-342錶達水平分彆為0.0751、0.2022、0.0425、0.3064、0.1733和0.0090,APL緩解期miR-15b、miR-16、miR-107、miR-223和miR-342錶達水平高于APL初診者(P值均<0.05),而APL緩解期miR-181a錶達水平低于APL初診者(P<0.05).結論 特定miRNA參與APL細胞分化過程.
목적 연구급성조유립세포백혈병(APL)세포분화전후미소RNA(miRNA,miR)표체변화.방법 채용전반식유갑산(ATRA)화삼양화이신(As2O3)유도APL세포계NB4 세포분화,서씨-희모살염색관찰세포형태,류식세포술검측세포표면표지CD11b적표체,용실시정량RT-PCR검측miRNA표체보miR-15b、miR-16、miR-34a、miR-107、miR-124a、miR-146、miR-155、miR-181a、miR-223、miR-342、let7c등miRNA적표체수평,용2-△△Ct법계산miRNA상대표체수평.수집15례APL초진화15례APL완해기환자골수단개핵세포(MNC),용RT-PCR검측MNC miRNA 적표체수평,용2-△Ct법계산miRNA표체수평.결과 ATRA작용NB4세포96 h,miR-15b、miR-16、miR-107、miR-223、miR-342표체수평균현저상조,분별위대조조적3.40、4.22、5.41、20.03화5.29배,As2O3작용NB4세포96 h,miR-15b、miR-16、miR-107、miR-223、miR-342표체수평야현저상조,분별위대조조적3.62、2.49、2.58、4.27화1.94배,제miR-15b외,ATRA처리조miR-16、miR-107、miR-223화miR-342표체수평상조정도균고우As2O3처리조,기중우이miR-223위심.ATRA화As2O3치료후완해기APL환자miR-15b、miR-16、miR-107、miR-181a、miR-223화miR-342표체수평(2-△Ct치)분별위0.4137、0.6367、0.1260、0.0522、0.6611화0.0280,이APL초진환자miR-15b、miR-16、miR-107、miR-181a、miR-223、miR-342표체수평분별위0.0751、0.2022、0.0425、0.3064、0.1733화0.0090,APL완해기miR-15b、miR-16、miR-107、miR-223화miR-342표체수평고우APL초진자(P치균<0.05),이APL완해기miR-181a표체수평저우APL초진자(P<0.05).결론 특정miRNA삼여APL세포분화과정.
Objective To study the expression profile of microRNAs in acute
promyelocytic leukemia (APL) cells during differentiation. Methods Differentiation of APL cell line NB4 cells was induced by all-trans retinoic acid (ATRA) and arsenic trioxide (As2O3). Morphological and immunological assay was performed by Wright-Giemsa staining and flow-cytometric analysis of CD11b surface expression. During in vitro NB4 differentiation
induced by ATRA and As2O3, microRNA expression profiles (miR-15b, miR-16, miR-34a, miR-107, miR-124a, miR-146, miR-155, miR-181a, miR-223, miR-342, let7c) were detected by real time RT-PCR, and the relative expression level of microRNAs were quantitatively analyzed by using 2-△△Ct, and compared with that of control group. Meanwhile, the microRNA expression profiles were also detected in 15 newly diagnosed APL patients and 15 complete remission (CR) APL cases by real time RT-PCR, and the relative expression level of microRNA was quantitated
by using 2-△Ct, and compared with that of control group (newly diagnosed APL as control group). These data were expressed as ±s, and differences between groups were examined using t test. P<0.05 was considered statistically significant. Results The expression levels of miR-15b, miR-16, miR-107, miR-223 and miR-342 in NB4 differentiation group were obviously up-regulated (3.40, 4.22, 5.41, 20.03 and 5.29 folds higher in ATRA treated NB4 cells than that of control group respectively, and 3.62, 2.49, 2.58, 4.27 and 1.94 folds higher in AS2O3 treated NB4 cells than that of control group respectively), except for miR-15b, the expression levels of miR-16, miR-107, miR-223 and miR-342 in ATRA treated group was significantly higher than that in As2O3 treated group. The relative expression levels of miR-15b, miR-16, miR-107, miR-181a, miR-223 and miR-342 were 0.4137, 0.6367, 0.1260, 0.0522, 0.6611, 0.0280 in APL CR group, and 0.0751, 0.2022, 0.0425, 0.3064, 0.1733, 0.0090 in newly diagnosed APL group, respectively. The expression level of miR-15b, miR-16, miR-107, miR-223 and miR-342 in APL CR group were significantly upregulated compared with that of newly diagnosed APL groups (P<0.05), while the expression level of miR-181a was significantly downregulated (P<0.05). Conclusion Specific expression of microRNA profiles is a key contributing factor in the differentiation of APL.
