|
|
Apoptosis of acute myeloid leukemia HL-60 cells induced by CDK inhibitor SNS-032 and its molecular mechanisms |
HAN Yan-xia1,2, YOU Liang-shun1, LIU Hui1, MAO Li-ping1, YE Xiu-jin1, QIAN Wen-bin1 |
1. Department of Hematology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China;
2. Department of Hematology, Jiaxing Second People's Hospital, Jiaxing 314000, China |
|
|
Abstract Objective: To investigate the effects of cycle-dependent kinase (CDK) inhibitor SNS-032 on apoptosis in human acute myeloid leukemia (AML) HL-60 cells and its molecular mechanisms. Methods: Cultured AML HL-60 cells were treated with various concentrations of SNS-032. Cell apoptosis was determined with flow cytometry;cell viability was measured by MTT assay; the profiles of microRNA expression of HL-60 cells were analyzed by microRNA microarray;the protein expressions of JAK2/STAT3 pathway were detected by Western blotting. Results: Apoptosis of AML HL-60 cells was induced by SNS-032; the rate of apoptosis was (5.9±1.7)%, (12.1±3.1)% and (59.4±3.6)% when HL-60 cells were treated with 0,100 and 200 nmol/L SNS-032. MicroRNA microarray analysis revealed that the levels of miR-30a, miR-183, miR-20b, miR-26b, miR-20a, miR-589, miR-107, miR-181a, miR-106a, miR-17 and miR-378c were down-regulated by SNS-032,whereas the levels of miR-320a and miR-H7* were up-regulated. Western blotting showed that SNS-032 strongly inhibited phosphorylation of STAT3 and protein expression of JAK2,C-MYC and MCL-1. Conclusion: CDK inhibitor SNS-032 can induce apoptosis of AML HL-60 cells, which is associated with the inhibition of MCL-1,C-MYC and JAK2/STAT3, and down-regulation of miR-17-92 family.
|
Received: 05 September 2014
Published: 25 March 2015
|
|
细胞周期蛋白依赖激酶抑制剂诱导HL-60细胞凋亡及分子机制研究
目的:探讨细胞周期蛋白依赖激酶(CDK)抑制剂SNS-032诱导人急性髓系白血病HL-60细胞凋亡的效应及可能的机制。 方法:以CDK抑制剂SNS-032作用于HL-60细胞株,实验细胞分为对照组、SNS-032组、白细胞介素(IL)-6组和SNS-032+IL-6组。MTT法检测细胞存活率,流式细胞术检测细胞凋亡, microRNA芯片技术分析细胞microRNA的表达谱差异,蛋白质印迹法检测JAK/STAT3相关信号通路蛋白的表达。 结果:SNS-032可降低细胞存活率,诱导HL-60细胞凋亡,对照组、100nmol/L和200nmol/L的SNS-032干预组细胞凋亡率分别为(5.9±1.7)%、(12.1±3.1)%和(59.4±3.6)%。microRNA芯片分析结果显示,SNS-032显著下调HL-60细胞miR-30a、miR-183、miR-20b、miR-26b、miR-20a、miR-589、miR-107、miR-181a、miR-106a、miR-17和miR-378c的表达水平,显著上调miR-320a的表达水平。蛋白质印迹法显示SNS-032可抑制STAT3的磷酸化和JAK2、MCL-1、C-MYC蛋白的表达。作为JAK/STAT3通路的激活剂,IL-6联合SNS-032不能逆转后者对HL-60细胞的杀伤作用(P>0.05),也不能逆转SNS-032对JAK2蛋白表达和磷酸化STAT3的抑制作用。结论:SNS-032能显著诱导人急性髓系白血病 HL-60细胞发生凋亡,其机制可能与抑制JAK2和STAT3磷酸化、抑制MCL-1和C-MYC及与之相关的miR-17-92基因簇表达水平有关。
关键词:
白血病,髓样,急性,
微RNAs,
细胞周期蛋白质依赖激酶类/生物合成,
Janus激酶类,
STAT3转录因子,
蛋白酪氨酸激酶类/代谢,
信号传导,
HL-60细胞,
细胞凋亡,
肿瘤细胞,培养的
|
|
[1] ASGHAR U, WITKIEWICZ A K, TURNER N C, et al. The history and future of targeting cyclin-dependent kinases in cancer therapy [J]. Nat Rev Drug Discov, 2015, 14(2):130-146.
[2] KUMAR S K, LAPLANT B, CHNG W J, et al. Dinaciclib, a novel CDK inhibitor, demonstrates encouraging single-agent activity in patients with relapsed multiple myeloma [J]. Blood, 2015, 125(3):443-448.
[3] DICKSON M A. Molecular pathways:CDK4 inhibitors for cancer therapy [J]. Clin Cancer Res, 2014, 20(13):3379-3383.
[4] CHEN R, WIERDA W G, CHUBB S, et al. Mechanism of action of SNS-032, a novel cyclin-dependent kinase inhibitor, in chronic lymphocytic leukemia [J]. Blood, 2009, 113(19):4637-4645.
[5] WALSBY E, LAZENBY M, PEPPER C, et al. The cyclin-dependent kinase inhibitor SNS-032 has single agent activity in AML cells and is highly synergistic with cytarabine [J]. Leukemia, 2011, 25(3):411-419.
[6] LEE Y, AHN C, HAN J, et al. The nuclear RNase III Drosha initiates microRNA processing [J]. Nature, 2003, 425(6956):415-419.
[7] DENLI A M, TOPS B B, PLASTERK R H, et al. Processing of primary microRNAs by the Microprocessor complex [J]. Nature, 2004, 432(7014):231-235.
[8] LI Z, LU J, SUN M, et al. Distinct microRNA expression profiles in acute myeloid leukemia with common translocations [J]. Proc Natl Acad Sci USA, 2008, 105(40):15535-15540.
[9] DIXON-MCIVER A, EAST P, MEIN C A, et al. Distinctive patterns of microRNA expression associated with karyotype in acute myeloid leukaemia [J]. PLoS One, 2008, 3(5):e2141.
[10] JIN H Y, LAI M, XIAO C. microRNA-17~92 is a powerful cancer driver and a therapeutic target [J]. Cell Cycle, 2014, 13(4):495-496.
[11] XIANG J, WU J. Feud or friend? The role of the miR-17-92 cluster in tumorigenesis [J]. Curr Genomics, 2010, 11(2):129-135.
[12] CHANG Q, BOURNAZOU E, SANSONE P, et al. The IL-6/JAK/STAT3 feed-forward loop drives tumorigenesis and metastasis [J]. Neoplasia, 2013, 15(7):848-862. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|