促红细胞生成素对达卡巴嗪抗小鼠黑色素瘤细胞活性的调节作用

doi:10.3971/j.issn.1000-8578.2012.12.008

促红细胞生成素对达卡巴嗪抗小鼠黑色素瘤细胞活性的调节作用

Modulatable Effect of Erythropoietin on Anticancer Activity of Dacarbazinein Murine Melanoma Cells

摘要

摘要: 目的检测小鼠黑色素瘤细胞株B16促红细胞生成素受体(Erythropoietin receptor,EPOR)的表达，探讨促红细胞生成素(Erythropoietin,EPO)对达卡巴嗪抗B16细胞活性的影响及与Bcl-2家族蛋白表达的关系。方法采用RT-PCR、免疫荧光和激光共聚焦显微镜、Western blot等方法检测B16 细胞EPOR mRNA和蛋白的表达。MTT法和克隆形成试验检测EPO与达卡巴嗪联合应用时对B16细胞增殖的影响。Western blot法检测EPO对B16细胞Bcl-2、Bcl-xL、Bax蛋白表达的影响。结果B16细胞表达EPOR mRNA和蛋白。EPO能有效减少达卡巴嗪诱导B16细胞的凋亡（P<0.05），EPO作用后，B16细胞Bcl-2、Bcl-xL蛋白表达水平明显增加（P<0.05），Bax蛋白表达水平未见明显变化。结论B16细胞表达EPOR，EPO可调节化疗药对B16细胞的作用，其作用机制可能与其影响Bcl-2家族蛋白表达有关。

Abstract: Objective To investigate the expression of EPOR and the role of erythropoietin(EPO)in the melanoma cell line B16 proliferation and its relation to the expression of Bcl-2 family proteins. Methods Expression of EPOR mRNA and protein was detected by RT-PCR,immunofluorescen laser scanning microscope(LSM)and Western blot.Effects of EPO and combining with dacarbazine on B16 cells proliferation was measured by MTT methods and colony forming assay.Expressions of Bcl-2,Bcl-xL and Bax proteins were analyzed by Western blot. Results EPOR mRNA and protein can be expressed in B16 cells.Pretreatment of B16 cells with EPO can significantly increase the resistance to dacarbazine treatment（P<0.05）.Expression of both Bcl-2 and Bcl-xL proteins in B16 cell increased（P<0.05）,while Bax protein remained unchanged. Conclusion B16 cells can express EPOR.EPO can regulate antineoplastic activity of dacarbazine and its mechanism was probably related to that EPO can regulate the expression of bcl-2 family proteins.

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