艾拉莫德对人多发性骨髓瘤细胞株RPMI8226增殖和凋亡的影响
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摘要
目的探讨艾拉莫德对人多发性骨髓瘤(multiple myeloma,MM) RPMI8226细胞增殖和凋亡的影响。方法实验分组为:对照组(0μg/ml艾拉莫德),10μg/ml艾拉莫德处理组,20μg/ml艾拉莫德处理组,30μg/ml艾拉莫德处理组。艾拉莫德(10,20,30μg/ml)作用于人多发性骨髓瘤细胞株RPMI8226细胞24 h,采用MTT法检测RPMI8226细胞活力,Hoechst 33258染色检测凋亡细胞;荧光定量PCR检测Notch1 m RNA表达水平; Western blot检测Bax和Notch1蛋白的表达水平。结果10,20和30μg/ml的艾拉莫德处理24 h,可降低RPMI8226细胞活力,并具剂量依赖性(P <0. 05)。艾拉莫德(10,20,30μg/ml)处理RPMI8226细胞24 h,细胞核出现固缩、裂解现象。10,20和30μg/ml的艾拉莫德处理细胞24 h可显著上调RPMI8226细胞的Bax蛋白表达水平(P <0. 05),并显著下调Notch1 m RNA和蛋白水平(P <0. 05)。结论艾拉莫德可抑制人多发性骨髓瘤细胞株RPMI8226细胞增殖并促进细胞凋亡,其作用机制可能与其抑制Notch通路的活化有关。
Objective To explore the effects of iguratimod on cell proliferation and apoptosis of human multiple myeloma cell line RPMI8226 cells. Methods The experiment was divided into control group( 0 μg/ml iguratimod),10 μg/ml iguratimod group,20 g/ml iguratimod group,and 30 μg/ml iguratimod group. After treated RPMI8226 cells with different concentrations of iguratimod for 24 h,the cell viability was measured using MTT assay,the morphology of apoptotic cell was observed under a fluorescence microscope after Hoechst 33258 staining,the expression level of Notch1 m RNA was detected by quantitative real-time PCR,and the expression levels of Bax and Notch1 protein were detected by Western blot. Results Compared with control group,iguratimod significantly reduced the viability of RPMI8226 cells in a dose dependent manner( P < 0. 05). The cell nuclei showed pyknosis and lysis in RPMI8226 cells treated with iguratimod( 10,20,and 30 μg/ml) for 24 h,and the expression level of Bax protein was upregulated( P < 0. 05). In addition,the levels of Notch1 m RNA as well as protein in RPMI8226 cells were markedly ameliorated after treatment with iguratimod( 10,20,and 30 μg/ml) for 24 h( P < 0. 05). Conclusion Iguratimod can inhibit cell proliferation and induce cell apoptosis of human myeloma cell line RPMI8226,which may be related to inhibition of Notch pathway activation.
Objective To explore the effects of iguratimod on cell proliferation and apoptosis of human multiple myeloma cell line RPMI8226 cells. Methods The experiment was divided into control group( 0 μg/ml iguratimod),10 μg/ml iguratimod group,20 g/ml iguratimod group,and 30 μg/ml iguratimod group. After treated RPMI8226 cells with different concentrations of iguratimod for 24 h,the cell viability was measured using MTT assay,the morphology of apoptotic cell was observed under a fluorescence microscope after Hoechst 33258 staining,the expression level of Notch1 m RNA was detected by quantitative real-time PCR,and the expression levels of Bax and Notch1 protein were detected by Western blot. Results Compared with control group,iguratimod significantly reduced the viability of RPMI8226 cells in a dose dependent manner( P < 0. 05). The cell nuclei showed pyknosis and lysis in RPMI8226 cells treated with iguratimod( 10,20,and 30 μg/ml) for 24 h,and the expression level of Bax protein was upregulated( P < 0. 05). In addition,the levels of Notch1 m RNA as well as protein in RPMI8226 cells were markedly ameliorated after treatment with iguratimod( 10,20,and 30 μg/ml) for 24 h( P < 0. 05). Conclusion Iguratimod can inhibit cell proliferation and induce cell apoptosis of human myeloma cell line RPMI8226,which may be related to inhibition of Notch pathway activation.
引文
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[21]Xia S,Zhang X,Li C,et al.Oridonin inhibits breast cancer growth and metastasis through blocking the Notch signaling[J].Saudi Pharm J,2017,25(4):638-643.
[2]Baris D,Brown LM,Andreotti G,et al.Epidemiology of Multiple Myeloma[M].New York:Springer,2013:547-563.
[3]Okamura K,Yonemoto Y,Okura C,et al.Efficacy of the clinical use of iguratimod therapy in patients with rheumatoid arthritis[J].Mod Rheumatol,2015,25(2):235-240.
[4]Wang X,Ma C,Li P,et al.Effects of iguratimod on the levels of circulating regulators of bone remodeling and bone remodeling markers in patients with rheumatoid arthritis[J].Clin Rheumatol,2017,36(6):1369-1377.
[5]Alzrigat M,Parraga AA,Jernberg-Wiklund H.Epigenetics in multiple myeloma:from mechanisms to therapy[J].Semin Cancer Biol,2018,51:101-115.
[6]Mahindra A,Hideshima T,Anderson KC.Multiple myeloma:biology of the disease[J].Blood Rev,2010,24 Suppl:S5-11.
[7]Luo Q,Sun Y,Liu W,et al.A novel disease-modifying antirheumatic drug,iguratimod,ameliorates murine arthritis by blocking IL-17 signaling,distinct from methotrexate and leflunomide[J].J Immunol,2013,191(10):4969-4978.
[8]Okamura K,Yonemoto Y,Okura C,et al.Efficacy of the clinical use of iguratimod therapy in patients with rheumatoid arthritis[J].Mod Rheumatol,2015,25(2):235-240.
[9]Li J,Mao H,Liang Y,et al.Efficacy and safety of iguratimod for the treatment of rheumatoid arthritis[J].Clin Dev Immunol,2013,2013:310628.
[10]Sun X,Liao W,Wang J,et al.CSTMP induces apoptosis and mitochondrial dysfunction in human myeloma RPMI8226 cells via CHOP-dependent endoplasmic reticulum stress[J].Biomed Pharmacother,2016,83:776-784.
[11]Lang T,Nie Y.Mi R-148a participates in the growth of RPMI8226multiple myeloma cells by regulating CDKN1B[J].Biomed Pharmacother,2016,84:1967-1971.
[12]Du S,Qin W,Leng H,et al.Construction of a recombinant lentivirus-mediated sh RNA expression vector targeting the human PSMD10 gene and validation of RNAi efficiency in RPMI8226multiple myeloma cells[J].Oncol Rep,2017,38(2):809-818.
[13]Sakamoto T,Ishii Y,Shiba H,et al.Inhibitory effect of antirheumatic drug iguratimod for hepatocellular carcinogenesis by inhibition of serum interleukin-8 production[J].Anticancer Res,2016,36(7):3301-3306.
[14]Kopan R,Ilagan MX.The canonical Notch signaling pathway:unfolding the activation mechanism[J].Cell,2009,137(2):216-233.
[15]Lai EC.Notch signaling:control of cell communication and cell fate[J].Development,2004,131(5):965-973.
[16]Capaccione KM,Pine SR.The Notch signaling pathway as a mediator of tumor survival[J].Carcinogenesis,2013,34(7):1420-1430.
[17]李美玲,陈美琼,张鹏,等.靶向沉默Notch1基因对骨髓瘤细胞增殖的影响[J].中国实验血液学杂志,2017,25(06):1707-1712
[18]Zeng Y,Yin B,Wang X,et al.Effects of the Notch1 signaling pathway on human lung cancer A549 cells[J].Exp Lung Res,2017,43(4-5):208-216.
[19]Skrtic A,Korac P,Kristo DR,et al.Immunohistochemical analysis of NOTCH1 and JAGGED1 expression in multiple myeloma and monoclonal gammopathy of undetermined significance[J].Hum Pathol,2010,41(12):1702-1710.
[20]Iso T,Kedes L,Hamamori Y.HES and HERP families:multiple effectors of the Notch signaling pathway[J].J Cell Physiol,2003,194(3):237-255.
[21]Xia S,Zhang X,Li C,et al.Oridonin inhibits breast cancer growth and metastasis through blocking the Notch signaling[J].Saudi Pharm J,2017,25(4):638-643.