miR205-3p靶向MDM2调控乳腺癌血管内皮细胞增殖的研究  

MiR205-3p regulates proliferation of vascular endothelial cells in breast cancer through inhibiting MDM2 expression

作　　者：周少丞 季晓春[1] 滕伟峰 张佳男 毛琦淇 ZHOU Shaocheng;JI Xiaochun;TENG Weifeng;ZHANG Jianan;MAO Qiqi(Department of Thyroid and Breast Surgery,Ningbo Medical Center)

机构地区：[1]宁波市医疗中心李惠利医院甲乳外科,315040

出　　处：《浙江医学》2021年第15期1614-1618,1623,I0004,共7页Zhejiang Medical Journal

基　　金：宁波市科技计划项目(202003N4022)。

摘　　要：目的分析miR205-3p通过靶向鼠双微体2基因(MDM2)调控乳腺癌血管内皮细胞增殖的作用机制,探讨乳腺癌临床治疗可行的分子靶点。方法采用乳腺癌细胞(MDA-MB-231)皮下移植瘤组织为试验样本,通过免疫荧光实验检测移植瘤组织中是否有人源性肿瘤血管内皮细胞的存在,利用血管内皮细胞纯化技术得到移植瘤中的血管内皮细胞(即乳腺癌血管内皮细胞)。采用显微镜下观察细胞形态、内皮细胞成管实验及吞噬实验,验证血管内皮细胞的形态及生物学特性。检测乳腺癌血管内皮细胞和人脐静脉内皮细胞的miR205-3p的表达是否有差异,采用生物信息学分析方法得到miR205-3p的靶基因。通过改变miR205-3p的表达,研究靶基因的蛋白表达水平的改变。采用体外增殖实验及体内成瘤实验观察miR205-3p对乳腺癌血管内皮细胞及乳腺癌移植瘤增殖能力的影响。结果乳腺癌血管内皮细胞具有内皮细胞功能。相比较人脐静脉内皮细胞,miR205-3p在乳腺癌血管内皮细胞中表达降低(P<0.05)。利用生物信息学分析得到miR205-3p的靶基因为MDM2,提高miR205-3p的表达水平后可降低MDM2蛋白的表达(P<0.05),并在体内及体外水平都可影响乳腺癌血管内皮细胞的增殖能力。结论miR205-3p的表达水平升高可有效降低MDM2的表达,抑制肿瘤血管内皮细胞的增殖,这为抗肿瘤血管生成作用的研究及靶向药物开发提供了新的思路。Objective To investigate the effects of miRNA205-3p on proliferation of vascular endothelial cells in breast cancer tissue and its molecular mechanism.Methods Human breast cancer MDA-MB-231 cells were transplanted in nude mice.The vascular endothelial cells in transplanted breast cancer tissue were identified with immunofluorescence assay and their biological functions were tested with endothelial cell formation assay and phagocytosis assay.The expression of miR205-3p in endothelial cells was detected with qRT-PCR and compared with that in human umbilical vein endothelial cells.Bioinformatics analysis and luciferase reporter gene experiment were used to determine the target genes of miR205-3p.The effect of miR205-3p on the protein expression of target genes was determined.The effects of miR205-3p on the proliferation of breast cancer vascular endothelial cells and breast cancer were detected in vitro and in vivo.Results Compared with human umbilical vein endothelial cells,the expression of miR205-3p in breast cancer-endothelial cells was significantly decreased(P<0.05).Bioinformatics analysis and luciferase reporter gene experiment suggested that the targeting gene of miR205-3p was MDM2.Increasing the expression level of miR205-3p reduced the expression of MDM2 at the protein level(P<0.05),and affected proliferation ability of breast cancer-endothelial cells in vivo and in virto.Conclusion The miR205-3p can inhibit the proliferation of tumor vascular endothelial cells through down-regulation of MDM2 expression,which might be a novel target for anti-tumor angiogenesis treatment.

关 键 词：miR205-3p 乳腺癌血管内皮细胞 CD105 鼠双微体2 

分 类 号：R737.9[医药卫生—肿瘤]