长链非编码DLEU1靶向miR-7的表达通过BCR信号通路调控儿童白血病细胞的生物学行为
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摘要
目的:研究长链非编码DLEU1调控儿童白血病细胞生物学行为的作用及其机制。方法:qPCR检测不同白血病细胞中DLEU1的表达情况;双荧光素酶报告基因检测DLEU1与miR-7的相互作用;划痕愈合实验检测过表达DLEU1后白血病细胞迁移能力的变化;Transwell侵袭实验检测过表达DLEU1后白血病细胞侵袭能力的变化;划痕愈合实验和Transwell侵袭实验检测过表达DLEU1后miR-7对白血病细胞迁移和侵袭能力的影响;Western blotting检测过表达DLEU1后BCR信号通路蛋白的表达情况。结果:在白血病细胞KG-1中DLEU1表达水平最低;DLEU1能与miR-7的3'UTR特异性结合;过表达DLEU1可以抑制白血病细胞侵袭和迁移能力;过表达DLEU1后,过表达miR-7可以促进白血病细胞侵袭和迁移能力;过表达DLEU1后BCR通路蛋白表达相应下调。结论:DLEU1可以靶向调节miR-7通过BCR信号通路调控白血病细胞的侵袭和迁移能力。
Objective:To investigate the effect and mechanism of long non-coding DLEU1 regulating the biological behavior of childhood leukemia cells.Methods:The expression of DLEU1 in different leukemia cells was detected by qPCR.Double luciferase reporter gene was used to detect the interaction between DLEU1 and miR-7.The changes of leukemia cells migration ability after silencing DLEU1 were detected by scratch healing experiment.Transwell invasion assay was used to detect the invasion ability of leukemia cells after silencing DLEU1.Scratch healing experiment and Transwell invasion test were used to detect the effect of miR-7 on the migration and invasion of leukemia cells after silencing DLEU1.Western blotting was used to detect the expression of BCR signaling pathway after silencing DLEU1.Results:The expression level of DLEU1 was the lowest in leukemia cells KG-1.DLEU1 could bind specifically to the 3' UTR of miR-7.Silence DLEU1 can inhibit leukemia cells invasion and migration ability.Overexpression of miR-7 can promote the invasion and migration of leukemia cells after silencing DLEU1.The expression of BCR pathway protein was down-regulated after silencing DLEU1.Conclusion:DLEU1 can regulate the invasion and migration of leukemic cells through the regulation of miR-7 through BCR signaling pathway.
Objective:To investigate the effect and mechanism of long non-coding DLEU1 regulating the biological behavior of childhood leukemia cells.Methods:The expression of DLEU1 in different leukemia cells was detected by qPCR.Double luciferase reporter gene was used to detect the interaction between DLEU1 and miR-7.The changes of leukemia cells migration ability after silencing DLEU1 were detected by scratch healing experiment.Transwell invasion assay was used to detect the invasion ability of leukemia cells after silencing DLEU1.Scratch healing experiment and Transwell invasion test were used to detect the effect of miR-7 on the migration and invasion of leukemia cells after silencing DLEU1.Western blotting was used to detect the expression of BCR signaling pathway after silencing DLEU1.Results:The expression level of DLEU1 was the lowest in leukemia cells KG-1.DLEU1 could bind specifically to the 3' UTR of miR-7.Silence DLEU1 can inhibit leukemia cells invasion and migration ability.Overexpression of miR-7 can promote the invasion and migration of leukemia cells after silencing DLEU1.The expression of BCR pathway protein was down-regulated after silencing DLEU1.Conclusion:DLEU1 can regulate the invasion and migration of leukemic cells through the regulation of miR-7 through BCR signaling pathway.
引文
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[24] Sun Xiaoyan,Li Jun,Sun Yanhong,et al.miR-7 reverses the resistance to BRAFi in melanoma by targeting EGFR/IGF-1R/CRAF and inhibiting the MAPK and PI3K/AKT signaling pathways[J].Oncotarget,2016,7(33):53558-53570.
[25] Zhang H,Luo XQ,Zhang P,et al.MicroRNA patterns associated with clinical prognostic parameters and CNS relapse prediction in pediatric acute leukemia[J].PLoS One,2009,4(11):e7826.
[2] Pizzitola I,Anjos-Afonso F,Rouault-pierre K,et al.Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid leukemia cells in vivo[J].Leukemia,2014,28(8):1596-1605.
[3] Nucera S,Giustacchini A,Boccalatte FA,et al.miRNA-126 orchestrates an oncogenic program in B cell precursor acute lymphoblastic leukemia[J].Cancer Cell,2016,29(6):905-921.
[4] Khouri IF,Anfossi S,Saliba RM,et al.Serum miRNA expression and allogeneic stem cell transplantation in patients with B chronic lymphocytic leukemia (CLL)[J].Blood,2014,124(21):5791-5791.
[5] Lee S,Yin Changhong,Ayello J,et al.Downregulation of DLEU1 significantly shortened survival in a Rituximab-treated DLEU1 knockout human burkitt lymphoma (BL) xenograft NSG mouse model:DLEU1 May act as a tumor suppressor gene in pediatric BL[J].Blood,2015,126(23):3656-3656.
[6] Cunningham F,Amode MR,Barrell D,et al.Ensembl 2015[J].Nucleic Acids Res,2015,43(D1):D662-D669.
[7] Oliveras-Ferraros C,Cufí S,Vazquez-Martin A,et al.Micro(mi)RNA expression profile of breast cancer epithelial cells treated with the anti-diabetic drug metformin:Induction of the tumor suppressor miRNA let-7a and suppression of the TGFβ-induced oncomiR miRNA-181a[J].Cell Cycle,2011,10(7):1144-1151.
[8] Zhang Hongyi,Cai Kai,Wang Jing,et al.MiR-7,inhibited indirectly by LincRNA HOTAIR,directly inhibits SETDB1 and reverses the EMT of breast cancer stem cells by downregulating the STAT3 pathway[J].Stem Cells,2014,32(11):2858-2868.
[9] Giles KM,Brown RA,Epis MR,et al.miRNA-7-5p inhibits melanoma cell migration and invasion[J].Biochem Biophys Res Commun,2013,430(2):706-710.
[10] Kovalchuk O,Filkowski J,Meservy J,et al.Involvement of microRNA-451 in resistance of the MCF-7 breast cancer cells to chemotherapeutic drug doxorubicin[J].Mol Cancer Ther,2008,7(7):2152-2159.
[11] Boeri M,Verri C,Conte D,et al.MicroRNA signatures in tissues and plasma predict development and prognosis of computed tomography detected lung cancer[J].Proc Natl Acad Sci USA,2011,108(9):3713-3718.
[12] Baccarani M,Deininger MW,Rosti GA,et al.European leukemianet recommendations for the management of chronic myeloid leukemia:2013[J].Blood,2013,122(6):872-884.
[13] Jabbour E,Kantarjian HM,Saglio G,et al.Early response with dasatinib or imatinib in chronic myeloid leukemia:3-year follow-up from a randomized phase 3 trial (DASISION)[J].Blood,2014,123(4):494-500.
[14] Lee S,Yin Changhong,Shah T,et al.Upregulation of DLEU1 significantly prolongs survival in a rituximab-treated DLEU1 knockout human Burkitt lymphoma (BL) xenograft NSG mouse model:DLEU1 May act as a tumor suppressor gene in pediatric BL[J].Cancer Res,2016,76(14):28-32.
[15] Wu Qian,Guo Li,Jiang Fei,et al.Analysis of the miRNA-mRNA-lncRNA networks in ER plus and ER-breast cancer cell lines[J].J Cell Mol Med,2015,19(12):2874-2887.
[16] Lee S,Yin Changhong,Ayello J,et al.DLEU1 significantly alters programmed cell death in chemoimmunotherapy-treated TALENs-induced DLEU1 knockout and DLEU1 overexpressing Burkitt Lymphoma (BL):DLEU1 may act as a tumor suppressor gene in pediatric BL[J].Cancer Res,2015,75(15):195-202.
[17] Feaver RE,Lawson M,Mackey AJ,et al.Assessment of donor and environmental risk factors in a human physiologically relevant model of nonalcoholic steatohepatitis (NASH)[C]//Hepatology.111 RIVER ST,HOBOKEN 07030-5774,NJ USA:WILEY-BLACKWELL,2015,62:660A-660A.
[18] Lee S,Luo W,Shah T,et al.The effects of DLEU1 gene expression in Burkitt lymphoma (BL):Potential mechanism of chemoimmunotherapy resistance in BL[J].Oncotarget,2017,8(17):27839.
[19] Hart JR,Weinberg MS,Morris KV,et al.MINCR is not a MYC-induced lncRNA[J].Proceedings of the National Academy of Sciences,2016,113(5):E496-E497.
[20] Cai JJ,Yang S,Zhang H,et al.Expression pattern of mRNA and non-coding RNA in ER+ and ER- breast cancer cells[J].Chinese Science Bulletin,2014,59(19):1861-1873.
[21] Nishioka C,Ikezoe T,Yang J,et al.BCR/ABL increases EZH2 levels which regulates XIAP expression via miRNA-219 in chronic myeloid leukemia cells[J].Leukemia Research,2016,45:24-32.
[22] Tansatit M.Applications of fluorescence in situ hybridization technology in malignancies[J].Cancer Cytogenetics:Methods and Protocols,2017,2(16):75-90.
[23] Balatti V,Pekarky Y,Rizzotto L,et al.miR deregulation in CLL[M].Adv Exp Med Biol,2013,792:309-325.
[24] Sun Xiaoyan,Li Jun,Sun Yanhong,et al.miR-7 reverses the resistance to BRAFi in melanoma by targeting EGFR/IGF-1R/CRAF and inhibiting the MAPK and PI3K/AKT signaling pathways[J].Oncotarget,2016,7(33):53558-53570.
[25] Zhang H,Luo XQ,Zhang P,et al.MicroRNA patterns associated with clinical prognostic parameters and CNS relapse prediction in pediatric acute leukemia[J].PLoS One,2009,4(11):e7826.