miR-361-5p 在三阴性乳腺癌（TNBC）中的过表达通过靶向 RQCD1 并抑制 EGFR/PI3K/Akt 通路抑制迁移和侵袭。
Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway.
机构信息
Breast Surgery, Affiliated Hospital of Hebei University of Engineering, Handan, Hebei, China.
出版信息
Bosn J Basic Med Sci. 2019 Feb 12;19(1):52-59. doi: 10.17305/bjbms.2018.3399.
Triple-negative breast cancer (TNBC) is the leading cause of cancer-related death in women. Previous studies indicated that miR-361-5p was downregulated in breast cancer, however, the exact effect of miR-361-5p on TNBC requires further investigation. In the present study, we investigated whether miR-361-5p can act as a tumor suppressor by targeting required for cell differentiation 1 homolog (RQCD1) and inhibiting epidermal growth factor receptor (EGFR)/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway in TNBC. The expression of miR-361-5p and RQCD1 was determined by quantitative reverse transcription PCR (qRT-PCR) and/or western blot in TNBC and the adjacent tissues. miR-361-5p mimics were constructed and transfected to TNBC cell line MDA-MB-231. Cells were divided into three groups: blank control group, miRNA mimic negative control (NC) group, and miR-361-5p mimics group. Expression of miR-361-5p, mRNA and protein expression of PI3K, Akt, EGFR, phosphorylated (p)-EGFR/PI3K/Akt, and protein expression of RQCD1 and matrix metallopeptidase 9 (MMP-9) in MDA-MB-231 were measured by qRT-PCR/western blot after transfection. Cell viability was determined by CCK-8 assay. Cell migration and invasion ability were evaluated by scratch and transwell assay, respectively. miR-361-5p target gene was determined by bioinformatics analysis and luciferase reporter assay. RQCD1 was identified as a target of miR-361-5p by TargetScan and confirmed by luciferase reporter assay. Downregulated miR-361-5p and upregulated RQCD1 were observed in TNBC tissues. Expression of EGFR, PI3K, Akt and MMP-9 was inhibited in cells treated with miR-361-5p mimics. Transfection of miR-361-5p mimics also inhibited the phosphorylation of EGFR, PI3K, and Akt. Suppressed cell viability, migration, and invasion was found in miR-361-5p mimics groups. Our results indicated that overexpression of miR-361-5p might act as a suppressor in TNBC by targeting RQCD1 to inhibit the EGFR/PI3K/Akt signaling pathway.
三阴性乳腺癌（TNBC）是女性癌症相关死亡的主要原因。先前的研究表明，miR-361-5p 在乳腺癌中下调，然而，miR-361-5p 对 TNBC 的确切影响需要进一步研究。在本研究中，我们通过靶向细胞分化所需的 1 同源物（RQCD1）和抑制表皮生长因子受体（EGFR）/磷酸肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）通路来研究 miR-361-5p 是否可以作为 TNBC 的肿瘤抑制因子。通过定量逆转录 PCR（qRT-PCR）和/或 Western blot 在 TNBC 和相邻组织中测定 miR-361-5p 和 RQCD1 的表达。构建 miR-361-5p 模拟物并转染至 TNBC 细胞系 MDA-MB-231。将细胞分为三组：空白对照组、miRNA 模拟物阴性对照组（NC）和 miR-361-5p 模拟物组。转染后，通过 qRT-PCR/Western blot 测量 MDA-MB-231 中 miR-361-5p、PI3K、Akt、磷酸化（p）-EGFR/PI3K/Akt 的 mRNA 和蛋白表达以及 RQCD1 和基质金属蛋白酶 9（MMP-9）的蛋白表达。通过 CCK-8 测定法测定细胞活力。通过划痕和 Transwell 测定分别评估细胞迁移和侵袭能力。通过生物信息学分析和荧光素酶报告基因测定确定 miR-361-5p 的靶基因。TargetScan 鉴定 RQCD1 是 miR-361-5p 的靶基因，并通过荧光素酶报告基因测定证实。在 TNBC 组织中观察到 miR-361-5p 表达下调和 RQCD1 表达上调。用 miR-361-5p 模拟物处理的细胞中 EGFR、PI3K、Akt 和 MMP-9 的表达受到抑制。miR-361-5p 模拟物的转染也抑制了 EGFR、PI3K 和 Akt 的磷酸化。在 miR-361-5p 模拟物组中发现细胞活力、迁移和侵袭受到抑制。我们的结果表明，通过靶向 RQCD1 抑制 EGFR/PI3K/Akt 信号通路，过表达 miR-361-5p 可能在 TNBC 中作为抑制因子发挥作用。
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