ATRA诱导分化的NB4细胞浸润裸鼠肺组织模型的建立
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摘要
目的:建立全反式维甲酸(ATRA)诱导分化的NB4细胞浸润裸鼠肺组织模型,为探讨诱导分化综合征(DS)的发生机制、预防和治疗提供研究平台。方法:首先,取对数生长期的NB4细胞在ATRA诱导下、在RPMI 1640培养基中、在37℃和5%CO2的条件下培养,72 h后收获诱导分化后的NB4细胞接种裸鼠尾静脉。然后,30天后处死裸鼠取肺组织,用G显带方法检测裸鼠肺组织的染色体核型,HE染色观察裸鼠肺组织学,电子显微镜观察超微结构,RT-PCR法和FISH技术检测裸鼠肺组织PM L-RARa m RNA转录本及PML-RARa基因的表达。结果:实验组染色体核型符合NB4细胞特征,在组织学和超微结构上明显见到分化的NB4细胞浸润裸鼠肺组织,检测到PM L-RARa m RNA转录本和PML/RARa融合基因;对照组未见到分化的NB4细胞浸润裸鼠肺组织,PML-RARa m RNA转录本和PML/RARa融合基因检测阴性。结论:用ATRA诱导分化的NB4细胞成功的建立了浸润裸鼠肺组织模型,为探讨诱导分化综合征(DS)的发生机制、预防和治疗提供了研究平台。
Objective: To establish a nude mice model of differentiated NB4 infiltration into the lung tissue, and provide approach for the study on differentiation syndrome(DS). Methods: Firstly, logarithmic growth NB4 cells in RPMI 1640 medium containing 0.5μmol/L ATRA were cultured at 37℃ in a humidified 5% CO2 incubation and were induced differentiation by ATRA. Then the cells induced differentiation by ATRA were harverted at 72 h after incubation and were injected into the tail vein of BALB/c-nu female nude mice(SPF level, 6 weeks old). Then, after 30 days, the rats were sacrificed, the pathology, ultructure and chromosome karyotype of rats lung tissue were examined, PML-RARa m RNA and PML-RARa gene of rats lung tissue were measured by RT-PCR and by FISH. Results: In the trial group, the chromosome karyotype of lung tissue was the same with the NB4 cells specialized by ATRA. In the trial groups, the infiltration of NB4 cells specialized by ATRA into the lung tissue was observed by HE staining and electron microscope, but not in the control group. In the trial group, the PM L-RARa m RNA and PML-RARa gene expressed in the lung tissue, but not in the control group.Conclusions: A nude mice model of BN4 differentiated by ATRA infiltration into the lung tissue was established, which may contribute to the study on the pathogenesis, prevention and treatment of DS.
Objective: To establish a nude mice model of differentiated NB4 infiltration into the lung tissue, and provide approach for the study on differentiation syndrome(DS). Methods: Firstly, logarithmic growth NB4 cells in RPMI 1640 medium containing 0.5μmol/L ATRA were cultured at 37℃ in a humidified 5% CO2 incubation and were induced differentiation by ATRA. Then the cells induced differentiation by ATRA were harverted at 72 h after incubation and were injected into the tail vein of BALB/c-nu female nude mice(SPF level, 6 weeks old). Then, after 30 days, the rats were sacrificed, the pathology, ultructure and chromosome karyotype of rats lung tissue were examined, PML-RARa m RNA and PML-RARa gene of rats lung tissue were measured by RT-PCR and by FISH. Results: In the trial group, the chromosome karyotype of lung tissue was the same with the NB4 cells specialized by ATRA. In the trial groups, the infiltration of NB4 cells specialized by ATRA into the lung tissue was observed by HE staining and electron microscope, but not in the control group. In the trial group, the PM L-RARa m RNA and PML-RARa gene expressed in the lung tissue, but not in the control group.Conclusions: A nude mice model of BN4 differentiated by ATRA infiltration into the lung tissue was established, which may contribute to the study on the pathogenesis, prevention and treatment of DS.
引文
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[26]Amare PS,Baisane C,Saikia T,et al.Fluorescence in situ hybridization:a highly efficient technique of molecular diagnoeis and predication for disease course in patients with myeloid Leukemia[J].Cancer Genet Cytogenent,2001,131:125-134
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[30]Strocchio L,Gurnari C,Santoro N,et al.Arsenious trioxide and trans retinoic acid treatment fo chilhood acute promyelocytic leukemia[J].Br J Haematol,2018,126:1200-1206
[31]Zhu HH,Wu DP,Du X,et al.Oral arsenious plus retinoic acid versus intravenous arsenious plus retinoic acid for non-high-risk:a non-inferiority,randomised phase 3 trial[J].Lancet Oncol,2018,19:871-879
[2]张鹏,王树叶,胡龙虎,等.三氧化二砷注射液治疗急性早幼粒细胞白血病72例[J].中华血液学杂志,1992,17(2):58-60
[3]张鹏,王树叶,胡龙虎,等.三氧化二砷治疗急性早幼粒细胞白血病七年总结-附242例分析[J].中华血液学杂志,2000,02:10-13
[4]MI J Q,LI J M,Chen Z X,et al.How to manage acute promyelocytic leukemia[J].Leukemia,2012,26(8):1743-1751
[5]Larson RS,Tallman MS.Retinotic acid syndrome:manifestations,pathogenesis,and treatment[J].Best Prac Res Clin Haematol,2003,16:453-461
[6]Richard P,Werner B.Potentiation of retinoid-indeced diffefentiation of HL-60 and U937 cell linek by cytokines[J].Eur J Cancer,1991,27:53
[7]Wang H,Cao F,Li J,et al.Arsennic trioxide and mannitol for the treatment of acute promyelocytic leukemia relapse in the central nervous system[J].Blood,2014,124(12):1998-2000
[8]Stein E M,Tallman M S.Searching the central nervous system for relapsed acute promyelocytic leukemia[J].Leuke Lymphoma,2013,54(12):2584-2585
[9]Ohanian M,Rozovski U,Ravandi F,et al.Very high leves of lacate dehydrogenase at diagnosis predict central nervous system relapse in acute promyelocytic leukemia[J].Er J Haematol,2015,169(4):595-597
[10]Ninomiya M,Kiyoi H,Ito M,et al.Retinoic acid syndrome in NOD/scid mice induced by injecting an acute promyelocytic leukemia cell line[J].Leukemia,2004,18(3):442-448
[11]Zang C,Liu H,Ries C,et al.Enhanced migration of the acute promyelocytic leukemia cellline NB4 under in vitro conditions during short-term all-transretinoic asid treatment[J].J Cancer Res Clin Oncol,2000,126(1):33-40
[12]Dore AI,Santana-Lemos BA,Coser VM,et al.The association of ICAM-1 Exon 6(E469K)but not of ICAM-1 Exon 4(G241R)and PECAM-1 Exon 3(L125V)polymorphisms with the development of difeerentiaion syndrome in acute promyelocytic leukemia[J].J Leukoc Boil,2007,82(5):340-1343
[13]Tallman MS,Andersen JW,Schiffer CA,et al.Clinical descriptionof44 patients with acute promyelocytic leukemia who developed the retinoic acid syndrome[J].Blood,2000,95(1):90-95
[14]Luesink M,Penniings JL,Wissink WM,et al.Chemokine induction by all-trans retinoic acid and arsenic trioxide in acute promyelocytic leukemia:triggering The differentiation syndrome[J].Blood,2009,114(27):5512--5521
[15]Zhou J,Hu L H,Cui Z,et al.Interaction of SDF-1a and Cxcr4 plans an importentrole in pulmonary cellar infiltration in differentiation syndrome[J].Int J Hematol,2010,91:293-302
[16]R Kiesslism,PS Hochman,O Haller,et al.Evidence for a similer or common mechanism for reverse transcriptase polymerase chain reaction[J].British J Immuunol,1977,7:655-663
[17]Romijn J C.Growth of tumor cells with diffenrent sensitivities for murine natural killer cells in young and adult athymic mude mice[J].Exp Cell Biol,1985,53:24-31
[18]Federica Cavallo,Marco Forni,Carlo Riccardi,et al.Growth and Spread of Human Malignant T Lymphoblasts in Immunosuppressed Nude Mice:A Model for meningeal Leukemia[J].Blood,1992,80:1279-1283
[19]Lanotte M,Martin-Thonvenin V,Najman S,et al.NB4,a maturation inducible cell line with t(15;17)maker isolated from a human acute promyelocytic leukemia(M3)[J].Blood,1991,77:1080
[20]Marchwicka A,Cunningham A,Marcinkowska E,et al.Threapeutic use of selective synthetic ligands for retinotic acidreceptors:a patent review[J].Expert Opin Ther Pat,2016,26(8):957-971
[21]Gianni M,Koken MH,Chelbi-Alix MK,et al.Combined arsenic and retinoic acid treatment enhances differentiation and apoptosis in arsenic-resistant NB4 cells[J].Blood,1998,91(11):4300-4310
[22]Sanz MA,Grimwade D,Tallman MS,et al.Management of acute promyelocytic leukemia:r ecommendations from an expert panel on behanf of the European Leukemia Net[J].Blood,2009,113(9):1875-1891
[23]周晋,胡龙虎,崔喆,等.微重力旋转培养法体外拟拟维甲酸综合征及其与CXCR-4/SDF-1a表达关系[J].中华血液学杂志,2007,28(12):810-813
[24]Zhou J,Hu Long Hu,Cui Zhe,et al.Interaction of SDF-1a and Cxcr4plans an importentrole in pulmonary cellar infiltration in differentiation syndrome[J].Int J Hematol,2010,91:293-302
[25]周晋,胡龙虎,王桂芳,等.亚神酸诱导分化的急性早幼粒细胞白血病细胞浸润人体肺组织的体外模拟实验研究[J].中国实用内科杂志,2007,12:950-953
[26]Amare PS,Baisane C,Saikia T,et al.Fluorescence in situ hybridization:a highly efficient technique of molecular diagnoeis and predication for disease course in patients with myeloid Leukemia[J].Cancer Genet Cytogenent,2001,131:125-134
[27]Sharifi M,Salehi R,Gheisari Y,et al.Inhibition of micro RNAmiR-99a induces opoptosis and necrosis in human a human acute promyelocytic leukemia[J].Adv Biomed Res,2014,3:61
[28]ShaRifi M,Salehi R,Gheisari Y,et al.Inhibition of micro RNAmiR-99a induces opoptosis and inhibits cell proliferation in human acute promyelocytic leukemia through modulation of p63 expression[J].Mol Biol Rep,2014,41(5):2799-2808
[29]Testi AM,Pession A,Diverio D,et al.Risk-adapted treatment of acute promyelocytic leukemia:results from the international Consortium for Chilhood APL[J].Blood,2018,132:405-412
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