Department of Acupuncture, Taizhou Hospital Affiliated to Nanjing University of Chinese Medicine, Taizhou, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2847-2855. doi: 10.26355/eurrev_201904_17562.
The aim of this study was to explore the mechanism of miR-98-5p in influencing the malignant proliferation and metastasis capacities of breast cancer cells.
Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression level of miR-98-5p and GRB2-associated-binding protein 2 (Gab2) in breast cancer samples and cells. On-line target gene prediction software and Dual-Luciferase reporter assay were used to predict and verify the target genes of miR-98-5p, respectively. Cell proliferation was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Meanwhile, migration and invasion abilities, as well as the changes of epithelial-mesenchymal transition (EMT) after transfection were detected by transwell assay and Western blot assay, respectively.
Compared with adjacent non-tumor tissues and MCF-10A cells, the expression level of miR-98-5p in tumor tissues and MCF-7 cells was significantly declined, whereas Gab2 was markedly up-regulated. Besides, Gab2 was predicted as a target gene of miR-98-5p. Subsequent experiments indicated that the proliferation, migration, invasion and EMT of MCF-7 cells transfected with miR-98-5p were significantly inhibited. However, up-regulation of Gab2 attenuated the biological function of miR-98-5p on malignant abilities of breast cancer cells.
We showed that miR-98-5p served as anti-oncogene in breast cancer, which might provide a new therapeutic target for its treatment.
本研究旨在探讨 miR-98-5p 影响乳腺癌细胞恶性增殖和转移能力的机制。
采用实时定量聚合酶链反应（qRT-PCR）检测乳腺癌组织和细胞中 miR-98-5p 和生长因子受体结合蛋白 2 相关结合蛋白 2（Gab2）的表达水平。利用在线靶基因预测软件和双荧光素酶报告基因实验分别预测和验证 miR-98-5p 的靶基因。采用 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）法检测细胞增殖。同时，通过 Transwell 实验和 Western blot 实验检测转染后迁移和侵袭能力以及上皮间质转化（EMT）的变化。
与相邻非肿瘤组织和 MCF-10A 细胞相比，肿瘤组织和 MCF-7 细胞中 miR-98-5p 的表达水平明显降低，而 Gab2 明显上调。此外，Gab2 被预测为 miR-98-5p 的靶基因。后续实验表明，转染 miR-98-5p 的 MCF-7 细胞的增殖、迁移、侵袭和 EMT 明显受到抑制。然而，上调 Gab2 减弱了 miR-98-5p 对乳腺癌细胞恶性能力的生物学功能。
我们表明 miR-98-5p 在乳腺癌中作为抑癌基因发挥作用，可能为其治疗提供新的治疗靶点。
