miR-449a在乳腺癌裸鼠皮下移植瘤模型中的作用

发布时间：2018-01-20 13:04

本文关键词： miR-a 乳腺癌裸鼠移植瘤　出处：《西安交通大学学报(医学版)》2017年06期 　论文类型：期刊论文

【摘要】：目的探究敲低miR-449a对裸鼠皮下乳腺癌细胞移植瘤生长的影响。方法本实验通过采用乳腺癌MCF-7细胞系建立乳腺癌的裸鼠皮下移植瘤动物模型,将含有shRNA-miR-449a表达的质粒稳定转染MCF-7细胞,将筛选的稳转细胞采取多点注射的方法注入裸鼠皮下,采用阴性质粒为阴性对照,PBS作为空白对照;处理裸鼠后检测成瘤体积并计算肿瘤生长抑制率;Real-time PCR检测肿瘤组织miR-449a、Notch 1表达情况;Western blot检测移植瘤组织内的Notch 1、β-catenin和E-cadherin蛋白水平;免疫组织化学技术检测肿瘤组织内Notch1、E-cadherin表达水平。结果移植瘤形成过程中裸鼠未出现死亡,肿瘤体积检测后发现miR-449a敲低后能够显著抑制肿瘤的生长(P0.05);miR-449a在移植瘤组织内表达降低,同时Notch 1蛋白表达降低,β-catenin表达降低,而E-cadherin蛋白表达升高;免疫组化结果也显示Notch 1蛋白水平降低,E-cadherin蛋白的表达水平升高(P0.05)。结论敲低miR-449a可能是通过下调Notch 1、β-catenin蛋白水平,促进E-cadherin蛋白表达,从而抑制乳腺癌裸鼠皮下移植瘤的生长。
[Abstract]:Objective to explore the effect of silencing miR-449a on growth of breast cancer cells subcutaneously in nude mice. Methods using nude mice to establish the animal model of breast cancer breast cancer MCF-7 cell line through this experiment with shRNA-miR-449a expression plasmid transfected MCF-7 cells stably transfected cells by method of multiple point injection will be screening the injected into nude mice the skin, using negative plasmid as negative control, PBS as control treatment; nude mice detected after tumor volume and tumor growth inhibitory rate was calculated; Real-time PCR detection of tumor tissue miR-449a, Notch expression in 1; detection of transplanted tumor tissue Western in blot Notch 1, -catenin beta and E-cadherin protein levels were detected in tumor tissue; immunohistochemistry in Notch1, the expression level of E-cadherin. The results of transplantation tumor formation in nude mice without death, tumor volume was found after miR-449a knocking down Can inhibit the tumor growth (P0.05); the decreased expression of miR-449a in transplanted tumor tissues, while the expression of Notch 1 decreased, beta -catenin expression decreased, E-cadherin protein expression increased; immunohistochemistry results showed that Notch 1 protein levels decreased, increased the expression of E-cadherin protein (P0.05). Conclusion the knockdown of miR-449a may through the downregulation of Notch 1 beta, the protein level of -catenin, promote the expression of E-cadherin protein, thereby inhibiting subcutaneous breast cancer growth in nude mice.

【作者单位】：兰州大学第一医院乳腺科;
【基金】：甘肃省自然科学基金项目(No.2011Y0163,2012Y0633)~~
【分类号】：R-332;R737.9
【正文快照】： 微小RNA(microRNA,miRNA)是一类长度为18~25个核苷酸的广泛存在于真核细胞并由内源性基因编码的非编码单链RNA。miRNA参与细胞的多种生命活动,如细胞增殖和分化、细胞凋亡和胚胎的发育与调控等[1]。近年来的研究发现,miRNA对肿瘤的发生、发展和转移等均能发挥调节作用,主要是

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