miR-125a-5p通过靶向调控GAB2抑制乳腺癌细胞的迁移能力

发布时间：2018-02-27 10:34

本文关键词： 乳腺癌 miR-a-p GRB相关结合蛋白迁移　出处：《中国生物化学与分子生物学报》2017年09期 　论文类型：期刊论文

【摘要】：miR-125a-5p可负性调节GAB2表达,抑制胶质瘤细胞的侵袭和转移。本研究旨在证明miR-125a-5p抑癌作用的普遍性,即miR-125a-5p是否可通过靶向抑制GAB2抑制乳腺癌细胞的迁移。荧光素酶实验结果显示,miR-125a-5p可特异识别GAB2的3'-UTR,抑制报告酶的表达。荧光定量PCR结果揭示,与正常乳腺上皮细胞MCF-10A比较,miR-125a-5p在乳腺癌细胞MDA231和MCF-7中的表达明显降低;与迁移能力相对较低的MCF-7细胞比较,miR-125a-5p在迁移能力较高的MDA231细胞中的表达量更低。Western印迹结果证明,与空载体(对照)和anti-miR125a-5p转染细胞比较,转染miR-125a-5p明显抑制GAB2蛋白在乳腺癌细胞中的表达。Transwell结果显示,与空载体转染的对照细胞比较,转染miR-125a-5p的乳腺癌细胞穿过基质胶的细胞数明显减少;相反,转染anti-miR125a-5p的细胞穿过基质胶的细胞数却明显增多。上述结果提示,miR-125a-5p在正常的乳腺细胞中高表达,而在乳腺癌细胞中低表达,其表达水平与癌细胞的迁移能力和GAB2表达呈反向关系。本研究结果还提示,miR-125a-5p通过靶向负调控GAB2抑制乳腺癌细胞的迁移能力。总之,本研究证明,miR-125a-5p在肿瘤中发挥抑癌作用。
[Abstract]:MiR-125a-5p can negatively regulate the expression of GAB2, inhibit glioma cell invasion and metastasis. The purpose of this study is to prove the universality of the miR-125a-5p tumor suppressor role, namely whether miR-125a-5p can inhibit the migration of breast cancer cells to inhibit GAB2 by luciferase target. Experimental results show that miR-125a-5p can specifically recognize GAB2 3'-UTR, inhibit the expression of report enzymes. Fluorescence quantitative PCR results revealed that compared with normal mammary epithelial cells MCF-10A, expression of miR-125a-5p in MDA231 and MCF-7 in breast cancer cells decreased significantly; and the migration ability of MCF-7 cells is relatively low compared with lower dose.Western blot results prove that the expression of miR-125a-5p in high migration ability in MDA231 cells, and the empty vector (control) compared with anti-miR125a-5p transfected cells, the expression of.Transwell of transfected miR-125a-5p inhibited GAB2 protein in breast cancer cells display, and empty vector Compared with control cells transfected with the number of cells transfected with miR-125a-5p breast cancer cells through Matrigel was significantly reduced; on the contrary, the number of cells transfected with anti-miR125a-5p cells through Matrigel was significantly increased. These results indicated that miR-125a-5p was highly expressed in normal breast cells, while low expression in breast cancer cells, and its expression level cancer cell migration and GAB2 expression is negatively correlated. The results of this study also suggest that miR-125a-5p negatively regulated by GAB2 to target inhibition of breast cancer cell migration. In conclusion, this study demonstrated that miR-125a-5p in tumor suppressor role play.

【作者单位】：潍坊医学院病理学教研室;潍坊医学院附属医院;潍坊医学院医学研究实验中心;潍坊医学院护理学院;
【基金】：国家自然科学基金青年基金项目(No.81402389,No.81641111) 山东省自然科学基金(No.ZR2014HL077,No.ZR2015HL065) 山东省高等学校科技计划(No.J12LK03,No.J13LK03)资助~~
【分类号】：R737.9

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