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。
It has been known that miR-125a-5p inhibits the invasion and metastasis of glioma cells through negatively regulating GRB associated-binding protein 2（ GAB2）. To investigate its mechanism,luciferase reporter assays were carried out and results showed that miR-125a-5p directly targets GAB2.The qRT-PCR assays showed that the expression of miR-125a-5p in MDA231 and MCF-7 cells was lower than that of MCF-10 A. The expression of miR-125a-5p in highly invasive cells MDA231 was lower than that of minimally invasive cells MCF-7. Western blotting showed that the levels of GAB2 in breast cancercells transfected with miR-125a-5p were significanly lower than that in breast cancer cells transfected with empty vectors or anti-miR-125a-5p. Transwell assays showed that the number of cells through Matrigel in MDA231 cells transfected with miR-125a-5p was significantly reduced than control MDA231 cells. The number of cells through Matrigel was significantly increased in MCF-7 cells transfected with anti-miR-125a-5p than control MCF-7 cells. The results above suggested that the expression level of miR-125a-5p in normal human mammary epithelial cells was higher than that in breast cancer cells. The expression level of miR-125a-5p negatively correlated with the migration ability of cancer cells and the levels of GAB2. Our results also suggested that miR-125a-5p could inhibit breast cancer cell migration by negatively targeting GAB2. In conclusion,our studies propose that miR-125a-5p was a repressive gene in tumor.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2017年第9期925-930,共6页
Chinese Journal of Biochemistry and Molecular Biology
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~~
