本文關(guān)鍵詞： miR-a 乳腺癌 裸鼠 移植瘤　出處：《西安交通大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年06期 　論文類型：期刊論文

【摘要】：目的探究敲低miR-449a對裸鼠皮下乳腺癌細胞移植瘤生長的影響。方法本實驗通過采用乳腺癌MCF-7細胞系建立乳腺癌的裸鼠皮下移植瘤動物模型,將含有shRNA-miR-449a表達的質(zhì)粒穩(wěn)定轉(zhuǎn)染MCF-7細胞,將篩選的穩(wěn)轉(zhuǎn)細胞采取多點注射的方法注入裸鼠皮下,采用陰性質(zhì)粒為陰性對照,PBS作為空白對照;處理裸鼠后檢測成瘤體積并計算腫瘤生長抑制率;Real-time PCR檢測腫瘤組織miR-449a、Notch 1表達情況;Western blot檢測移植瘤組織內(nèi)的Notch 1、β-catenin和E-cadherin蛋白水平;免疫組織化學(xué)技術(shù)檢測腫瘤組織內(nèi)Notch1、E-cadherin表達水平。結(jié)果移植瘤形成過程中裸鼠未出現(xiàn)死亡,腫瘤體積檢測后發(fā)現(xiàn)miR-449a敲低后能夠顯著抑制腫瘤的生長(P0.05);miR-449a在移植瘤組織內(nèi)表達降低,同時Notch 1蛋白表達降低,β-catenin表達降低,而E-cadherin蛋白表達升高;免疫組化結(jié)果也顯示Notch 1蛋白水平降低,E-cadherin蛋白的表達水平升高(P0.05)。結(jié)論敲低miR-449a可能是通過下調(diào)Notch 1、β-catenin蛋白水平,促進E-cadherin蛋白表達,從而抑制乳腺癌裸鼠皮下移植瘤的生長。
[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.

【作者單位】： 蘭州大學(xué)第一醫(yī)院乳腺科;
【基金】：甘肅省自然科學(xué)基金項目(No.2011Y0163,2012Y0633)~~
【分類號】：R-332;R737.9
【正文快照】： 微小RNA(microRNA,miRNA)是一類長度為18~25個核苷酸的廣泛存在于真核細胞并由內(nèi)源性基因編碼的非編碼單鏈RNA。miRNA參與細胞的多種生命活動,如細胞增殖和分化、細胞凋亡和胚胎的發(fā)育與調(diào)控等[1]。近年來的研究發(fā)現(xiàn),miRNA對腫瘤的發(fā)生、發(fā)展和轉(zhuǎn)移等均能發(fā)揮調(diào)節(jié)作用,主要是

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