Breast Center, Affiliated Hospital of Qingdao University, Qingdao, P.R. China.
Eur Rev Med Pharmacol Sci. 2017 Aug;21(16):3617-3625.
Breast cancer is one of the most common malignant tumors in women worldwide. Considering the poor therapeutic effect of breast cancer, we are supposed to dissect the functioning mode of miR-509-5p on breast cancer cell growth and metastasis, providing therapeutic targets for breast cancer.
Quantitative Real-time PCR (qRT-PCR) assay was employed to detect miR-509-5p expression level. CCK8 assay and colony formation assay were incorporated to assess cell viability and proliferation capacities. Cell migration and invasion assay were performed to investigate metastasis capacity of breast cancer cells. Flow cytometry was used to identify cell apoptosis and cell cycle distribution. Protein levels were assessed by Western blotting assay. The target gene was predicted and verified by bioinformatics analysis and luciferase assay.
MiR-509-5p was obviously downregulated in breast cancer tissues when compared with pericarcinomatous tissues (n=76). Overexpressed miR-509-5p could attenuate breast cancer cell viability, proliferation, migration and invasion capacities, as well as promote cell apoptosis and induce cell cycle arrest at G0/G1 phase. Superoxide dismutase 2 (SOD2) was chosen as the target gene of miR-509-5p by bioinformatic analysis and Luciferase reporter assay. Moreover, restoration of SOD2 could rescue tumor suppression role of miR-509-5p on breast cancer tumorigenesis.
MiR-509-5p exerted tumor-suppressive effects on breast cancer progression and metastasis via targeting SOD2 in vitro, which provided an innovative and candidate target for diagnose and treatment of breast cancer.
乳腺癌是全球女性最常见的恶性肿瘤之一。鉴于乳腺癌的治疗效果不佳，我们应该剖析 miR-509-5p 对乳腺癌细胞生长和转移的作用模式，为乳腺癌提供治疗靶点。
采用实时定量聚合酶链反应（qRT-PCR）检测 miR-509-5p 的表达水平。CCK8 检测和集落形成实验用于评估细胞活力和增殖能力。细胞迁移和侵袭实验用于研究乳腺癌细胞的转移能力。流式细胞术用于鉴定细胞凋亡和细胞周期分布。Western blot 用于评估蛋白水平。通过生物信息学分析和荧光素酶报告实验预测和验证靶基因。
与癌旁组织相比（n=76），乳腺癌组织中 miR-509-5p 明显下调。过表达 miR-509-5p 可减弱乳腺癌细胞活力、增殖、迁移和侵袭能力，促进细胞凋亡，并诱导细胞周期停滞在 G0/G1 期。超氧化物歧化酶 2（SOD2）通过生物信息学分析和荧光素酶报告实验被选为 miR-509-5p 的靶基因。此外，SOD2 的恢复可以挽救 miR-509-5p 对乳腺癌肿瘤发生的抑制作用。
miR-509-5p 通过靶向 SOD2 在体外对乳腺癌的发生和转移发挥抑瘤作用，为乳腺癌的诊断和治疗提供了一个新的候选靶点。
