干扰人乳腺癌MDA-MB-231细胞 EPCR表达对内皮细胞增殖、迁移及脉管形成的影响
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国家自然科学青年基金(81101493);江苏省普通高校研究生科研创新课题(CXZZ13_0985)


Effects of low-expressed EPCR on proliferation,migration and angiogenesis of HUVECs in human breast cancer cell line MDA-MB-231
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    摘要:

    目的:通过观察干扰乳腺癌MDA-MB-231 细胞内皮蛋白C受体(endothelial protein C receptor,EPCR)表达对内皮细胞的增殖-迁移-脉管形成能力的变化,分析EPCR在肿瘤血管形成中的作用。方法:采用siRNA方法降低人乳腺癌MDA-MB-231 细胞EPCR的表达,通过RT-PCR及Western blot技术检测EPCR干扰效果,实验分为EPCR干扰组-无关序列组及未处理组。制备肿瘤条件培养基模拟肿瘤微环境培养 HUVECs 细胞,通过CCK-8法检测各组内皮细胞增殖能力,Transwell小室检测内皮细胞迁移能力,Matrigel检测内皮细胞脉管形成能力。结果:与未处理组及无关列序组相比,EPCR干扰组细胞的增殖-迁移及脉管形成能力均明显降低(P<0.05)。结论:干扰乳腺癌MDA-MB-231 细胞EPCR的表达能够抑制内皮细胞增殖-迁移及脉管形成能力,提示EPCR可能在肿瘤血管形成中起重要作用。

    Abstract:

    Objective:To observe the effect of endothelial protein C receptor(EPCR)on proliferation,migration and angiogenesis of human umbilical vein endothelial HUVECs cells,and to explore the role of EPCR in tumor angiogenesis. Methods:siRNA was performed to silence the expression of EPCR in human breast cancer cell line MDA-MB-231. The expression of EPCR in siRNA transfection was identified by reverse tansciption polymerse chain reaction(RT-PCR)and Western Blot. The experiment included three groups:the EPCR siRNA group,the irrelevant sequence group and the control group. HUVEC cells were cultured under tumor microenvironment,which was simulated by tumor conditioned medium. CCK-8 kit was performed to detect the endothelial cells proliferation,the transwell method was for detection of endothelial cell migration numbers,and matrigel in vitro small tube formation assay was performed to survey the state of tube formation. Results:The EPCR siRNA group showed lower cell proliferation activity,less number of cell migrats and tube formation than the other two groups (P﹤0.05). Conclusion:Low-expressed EPCR in human breast cancer cell line MDA-MB-231 can inhibit the proliferation,migration and angiogenesis of endothelial cells,which shows that EPCR may play an important role in tumor angiogenesis.

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刘冬梅,王庆苓,吴永平.干扰人乳腺癌MDA-MB-231细胞 EPCR表达对内皮细胞增殖、迁移及脉管形成的影响[J].南京医科大学学报(自然科学版),2015,(3):315-319

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  • 收稿日期:2014-07-24
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  • 在线发布日期: 2015-03-10
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