促红细胞生成素在人乳腺癌细胞株中的表达及作用

Expression and function of erythropoietin in human breast cancer cell lines

目的:通过检测重组人促红细胞生成素(recombinant human erythropoietin,rh-EPO)对乳腺癌细胞株增殖、凋亡及转移等生物学行为的影响,探讨EPO/EPO-R在乳腺癌发生发展中的作用。方法:运用RT-PCR、免疫细胞化学Envision法及蛋白质印迹法检测乳腺癌细胞株MCF-7和MDA-MB-231中EPO、EPO-R mRNA含量及蛋白表达;采用MTT比色试验、原位末端脱氧核糖核酸转移酶标记DNA链断端分析法(terminal deoxynucleotidyl transferase mediateddutp-Biotin nick end-labeling assay,TUNEL)、Transwell小室法观察不同浓度rh-EPO对这两种细胞增殖、凋亡和转移能力的影响。结果:MCF-7和MDA-MB-231细胞中均表达EPO及EPO-R;MTT比色试验和Transwell小室法显示,rh-EPO能增强两种细胞株的增殖活性和迁移侵袭能力,而且具有时间剂量依赖性;TUNEL法显示rh-EPO并没有抑制两株细胞凋亡的作用。rh-EPO对于MCF-7的作用效果强于MDA-MB-231细胞。结论:在不影响癌细胞凋亡的情况下,rh-EPO能够促进MCF-7和MDA-MB-231细胞的增殖活性,提高癌细胞的迁移和侵袭能力。

Objective： To detect recombinant human erythropoietin（rh-EPO） on the biological behavior of tumor cells such as proliferation,apoptosis and metastasis;and to explore the role of EPO and EPO receptor（EPO-R） in the development of breast cancer.Methods： The expression levels of EPO and EPOR in MCF-7 and MDA-MB-231 were detected by RT-PCR,immunocytochemistry and Western blotting.The proliferation abilities of MCF-7 and MDA-MB-231 cells interfered by rh-EPO in different concentration were detected by MTT assay.The Apoptosis of MCF-7 and MDA-MB-231 cells were evaluated by the terminal deoxynucleotidyl transferase mediated dutp-Biotin nick end-labeling assay（TUNEL）.The migration and invasion abilities of MCF-7 and MDA-MB-231 cells were investigated by transwell technique.Results： The results of RT-PCR and Western blotting showed that EPO and EPO-R were expressed in MCF-7 and MDA-MB-231.The MTT assay and invasion assay revealed that rh-EPO could enhance the capacity of proliferation and the ability of migration and Matrigel-invasion in vitro both in MCF-7 and MDA-MB-231 cell lines in a time and dose-dependent manner.In TUNEL,the results showed that rh-EPO failed to prevent apoptosis.Conclusion： rh-EPO could enhance the capacity of proliferation and the ability of migration and Matrigel-invasion in vitro both in MCF-7 and MDA-MB-231 cell lines,and the effect on MCF-7 cells was better than that on MDA-MB-231 cells,but could not induce cell apoptosis.