[全反式维甲酸和三氧化二砷诱导急性早幼粒细胞白血病细胞分化过程中的微小RNA表达谱]
[microRNAs expression profile in acute promyelocytic leukemia cell differentiation induced by all-trans retinoic acid and arsenic trioxide].
作者信息
Wu Yong, Li Xian-fang, Yang Jing-hui, Liao Xiao-ying, Chen Yuan-zhong
机构信息
Fujian Institute of Hematology, Key Laboratory of Hematology, Fujian Province, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
出版信息
Zhonghua Xue Ye Xue Za Zhi. 2012 Jul;33(7):546-51.
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(-ΔΔCt), 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(-ΔCt), and compared with that of control group (newly diagnosed APL as control group). These data were expressed as x(-) ± 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.
目的
研究急性早幼粒细胞白血病（APL）细胞在分化过程中微小RNA的表达谱。
方法
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。通过瑞氏-吉姆萨染色和CD11b表面表达的流式细胞术分析进行形态学和免疫学检测。在ATRA和As2O3诱导的体外NB4分化过程中，采用实时RT-PCR检测微小RNA表达谱（miR-15b、miR-16、miR-34a、miR-107、miR-124a、miR-146、miR-155、miR-181a、miR-223、miR-342、let7c），并使用2(-ΔΔCt)对微小RNA的相对表达水平进行定量分析，与对照组比较。同时，采用实时RT-PCR检测15例新诊断APL患者和15例完全缓解（CR）APL病例的微小RNA表达谱，使用2(-ΔCt)对微小RNA相对表达水平进行定量，与对照组（以新诊断APL为对照组）比较。这些数据以x(-)±s表示，组间差异采用t检验。P<0.05认为差异有统计学意义。
结果
NB4分化组中miR-15b、miR-16、miR-107、miR-223和miR-342的表达水平明显上调（ATRA处理的NB4细胞分别比对照组高3.40、4.22、5.41、20.03和5.29倍，As2O3处理的NB4细胞分别比对照组高3.62、2.49、2.58、4.27和1.94倍），除miR-15b外，ATRA处理组中miR-16、miR-107、miR-223和miR-342的表达水平明显高于As2O3处理组。APL CR组中miR-15b、miR-16、miR-107、miR-181a、miR-223和miR-342的相对表达水平分别为0.4137、0.
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