Btk调控MDR1逆转急性淋巴细胞白血病耐药的研究-东南大学学报医学版

Btk调控MDR1逆转急性淋巴细胞白血病耐药的研究

单位：1. 南京医科大学附属淮安第一医院血液科, 江苏淮安 223300;
2. 南京医科大学血液病重点实验室, 江苏南京 210029

关键词：急性淋巴细胞白血病依鲁替尼布鲁顿酪氨酸激酶 MDR1 P-糖蛋白转录因子κB 多药耐药

分类号：R733.71;R-33;R453

出版年·卷·期（页码）：2019·38·第二期（259-265）

摘要：

目的：通过观察Btk抑制剂依鲁替尼作用于急性淋巴细胞白血病（ALL）细胞株（Sup-B15）后细胞中MDR1 mRNA、P-gp蛋白的表达及细胞对化疗药物敏感性的变化，探讨逆转ALL多药耐药的有效方法。方法：（1）利用依鲁替尼处理Sup-B15，PCR法检测处理后细胞中MDR1 mRNA水平变化；免疫印迹法检测P-gp等蛋白水平变化；（2）利用化疗药物作用于依鲁替尼处理后的Sup-B15，CCK-8法检测细胞增殖。结果：（1）在Sup-B15中，依鲁替尼作用后的MDR1 mRNA表达较对照组降低；细胞中P-gp等蛋白表达量较对照组降低。（2）依鲁替尼处理后的Sup-B15在相同Ara-C、ADR浓度作用下，其细胞增殖率较对照组降低（P<0.05）。结论：（1）依鲁替尼可降低Sup-B15中MDR1 mRNA及P-gp蛋白的表达，其作用机制可能是通过抑制Btk、NFκB表达及生物学活性，从而抑制MDR1及P-gp的表达。（2）依鲁替尼增强ALL细胞对化疗药物的敏感性。

Objective:By targeting Btk in acute lymphoblastic leukemia (ALL) cells, to observe the expression of MDR1 and P-gp and the changes of cellular sensitivity to chemotherapy drugs of ALL cells, to further explore the role of Btk in the regulation of ALL multidrug resistance. Methods:(1) Sup-B15 were treated with ibrutinib, and the mRNA and the protein expression levels of Btk and MDR1 (P-gp) were detected by PCR and Western blot, respectively. (2)Sup-B15 were first treated with ibrutinib, and then with Ara-C or ADR respectively, the cell proliferation was detected by CCK-8 method. Results:(1) After being treated with ibrutinib, the mRNA and protein expression levels of MDR1 and P-gp were decreased as compared with the control group. (2) When Sup-B15 were treated with ibrutinib and followed by Ara-C or ADR respectively, the cell proliferation rate was lower than that of the control group (P<0.05). Conclusion:(1) Ibrutinib can decrease the expression of MDR1 mRNA and P-gp protein by inhibiting the expression of Btk and NFκB in ALL cells. (2) Ibrutinib can enhance the sensitivity of Sup-B15 cells to Ara-C and ADR.

参考文献：

[1] FYRBERG A,PETERSON C,KAGEDAL B,et al.Induction of fetal hemoglobin and ABCB1 gene expression in 9-beta-D-arabinofuranosylguanine-resistant MOLT-4 cells[J].Cancer Chemother Pharmacol,2011,68(3):583-591.
[2] LIN J,XING H Y,GONG Y P,et al.Multidrug resistant gene MDR1 contributes to development of imatinib-resistance in Ph (+) acute lymphoblastic leukemia cell line SUP-BS15RI[J].Journal of Sichuan University(Medical Science Edition),2012,43(5):657-660,665.
[3] RAHGOZAR S,MOAFI A,ABEDI M,et al.mRNA expression profile of multidrug-resistant genes in acute lymphoblastic leukemia of children, a prognostic value for ABCA3 and ABCA2[J].Cancer Biol Ther,2014,15(1):35-41.
[4] PONGSTAPOM W,PAKAKASAM S,CHAKSANGCHAICHOTE P,et al.MDR1 C3435T and C1236T polymorphisms:association with high-risk childhood acute lymphoblastic leukemia[J].Asian Pac J Cancer Prev,2015,16(7):2839-2843.
[5] ANSARⅡ M,SAUTY G,LABUDA M,et al.Polymorphism in multidrug resistance-associated protein gene 3 is associated with outcomes in childhood acute lymphoblastic leukemia[J].Pharmacogenomics J,2012,12(5):386-394.
[6] SU Y,LEE S H,SINKO P J.Inhibition of efflux transporter ABCG2/BCRP does not restore mitoxantrone sensitivity in irinotecan-selected human leukemia CPT-K5 cells:evidence for multifactorial multidrug resistance[J].Eur J Pharm Sci,2006,29(2):102-110.
[7] HUANG F F,WU D S,ZHANG L,et al.Inactivation of PTEN increases ABCG2 expression and the side population through the PI3K/Akt pathway in adult acute leukemia[J].Cancer Lett,2013,336(1):96-105.
[8] 江雪杰.成人急性淋巴细胞白血病中乳腺癌耐药蛋白基因表达及其临床意义[J].中国实验血液学杂志,2008,16(1):31-34.
[9] SHAH D S,KUMAR R.Steroid resistance in leukemia[J].World J Exp Med,2013,3(2):21-25.
[10] TAO S D,WANG C L,CHEN Y,et al.Btk and NFκB as prognostic biomarkers and potential therapeutic targets in B cell acute lymphoblastic leukemia[J].Int J Clin Exp Pathol,2016,9(8):7995-8005.
[11] DAVOUDI Z,AKBARZADEH A,RAHMATIYAMCHI M,et al.Molecular target therapy of AKT and NF-kB signaling pathways and multidrug resistance by specific cell penetrating inhibitor peptides in HL-60 cells[J].Asian Pac J Cancer Prev,2014,15(10):4353-4358.
[12] SOH S X,LIM J Y,HUANG J W,et al.Multi-agent chemotherapy overcomes glucocorticoid resistance conferred by a BIM deletion polymorphism in pediatric acute lymphoblastic leukemia[J].PLoS One,2014,9(8):e103435.
[13] VASSILEV A,OZER Z,NAVARA C,et al.Bruton's tyrosine kinase as an inhibitor of the Fas/CD95 death-inducing signaling complex[J].J Biol Chem,1999,274(3):1646-1656.
[14] LIN K,GLENN M A,HARRIS R J,et al.c-Abl expression in chronic lymphocytic leukemia cells:clinical and therapeutic implications[J].Cancer Res,2006,66(15):7801-7809.
[15] 陶善东,邓媛,何正梅,等.Btk及NFκB在急性髓系白血病细胞中的表达及其意义[J].中国实验血液学杂志,2013,21(1):25-28.
[16] 朱文艳,王志清,华海应.急性淋巴细胞性白血病13个月后初次缓解1例并文献复习[J].现代医学,2015,43(9):1163-1164.
[17] 邓媛,陶善东,张欣,等.PCI-32765和Bortezomib对B细胞肿瘤细胞系细胞增殖、凋亡的影响及其机制的研究[J].中国实验血液学杂志,2013,21:1178-1182.
[18] CHIEN W W,LE B C,RACHINEL N,et al.Differential mechanisms of asparaginase resistance in B-type acute lymphoblastic leukemia and malignant natural killer cell lines[J].Sci Rep,2015,5:8068.
[19] BUCHNER M,PARK E,GENG H,et al.Identification of FOXM1 as a therapeutic target in B-cell lineage acute lymphoblastic leukaemia[J].Nat Commun,2015,6:6471.
[20] ZHOU L,BAI H,WANG C,et al.microRNA125b promotes leukemia cell resistance to daunorubicin by inhibiting apoptosis[J].Mol Med Rep,2014,9(5):1909-1916.
[21] de OLIVEIRA J C,BRASSESCO M S,SCRIDELI C A,et al.MicroRNA expression and activity in pediatric acute lymphoblastic leukemia (ALL)[J].Pediatr Blood Cancer,2012,59(4):599-604.
[22] LIAO W,LIU X,PENG H,et al.The expression and functional study of miR-181a in pediatric acute lymphoblastic leukemia[J].Chinese Journal of Hematology,2015,36(1):53-57.
[23] NISHIOKA C,IKEZOE T,YANG J,et al.Downregulation of miR-217 Correlates with resistance of Ph(+) leukemia cells to ABL tyrosine kinase inhibitors[J].Cancer Sci,2014,105(3):297-307.
[24] FERNANDO T R,RODRIGUEZ-MALAVE N I,WATERS E V,et al.LncRNA expression discriminates karyotype and predicts survival in b-lymphoblastic leukemia[J].Mol Cancer Res,2015,13(5):839-851.
[25] SHISHIDO S,BONIG H,KIM Y M.Role of integrin alpha4 in drug resistance of leukemia[J].Front Oncol,2014,4:99.
[26] SHENG X,MITTELMAN S D.The role of adipose tissue and obesity in causing treatment resistance of acute lymphoblastic leukemia[J].Front Pediatr,2014,2:53.
[27] SAMUELS A L,BEESLEY A H,YADAV B D,et al.A pre-clinical model of resistance to induction therapy in pediatric acute lymphoblastic leukemia[J].Blood Cancer J,2014,4:e232.
[28] YEN C Y,CHIANG W F,LIU S Y,et al.Long-term stimulation of areca nut components results in increased chemoresistance through elevated autophagic activity[J].J Oral Pathol Med,2014,43(2):91-96.
[29] 王婷,陈芳源,韩洁英,等.CYP3A5参与急性白血病耐药机制的研究[J].中华血液学杂志,2003,24(6):286-289.
[30] KHARABI M B,GENG H,HURTZ C,et al.Mechanistic rationale for targeting the unfolded protein response in pre-B acute lymphoblastic leukemia[J].Proc Natl Acad Sci U S A,2014,111(21):E2219-E2228.
[31] COOLS J,MENTENS N,FURET P,et al.Prediction of resistance to small molecule FLT3 inhibitors:implications for molecularly targeted therapy of acute leukemia[J].Cancer Res,2004,64(18):6385-6389.
[32] 夏君燕.我国210例伊马替尼耐药慢性髓细胞白血病和Ph阳性急性淋巴细胞白血病ABL1基因突变特征[J].中华检验医学杂志,2012,35(1):17-22.
[33] TAMAI H,MIYAKE K,YAMAGUCHI H,et al.Resistance of MLL-AFF1-positive acute lymphoblastic leukemia to tumor necrosis factor-alpha is mediated by S100A6 upregulation[J].Blood Cancer J,2011,1:e38.
[34] BURKE M J,LAMBA J K,POUNDS S,et al.A therapeutic trial of decitabine and vorinostat in combination with chemotherapy for relapsed/refractory acute lymphoblastic leukemia[J].Am J Hematol,2014,89(9):889-895.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录