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hsa-miR-570-3p 通过靶向 CD274 阻断 PI3K/AKT/mTOR 信号通路对三阴性乳腺癌的保护作用。

Protective effect of hsa-miR-570-3p targeting CD274 on triple negative breast cancer by blocking PI3K/AKT/mTOR signaling pathway.

机构信息

Department of Medical Oncology, Fujian Cancer Hospital & Fujian Medical University Cancer Hospital, Fuzhou, Fujian Province, China.

Department of Pharmacy, Fujian Cancer Hospital & Fujian Medical University Cancer Hospital, Fuzhou, Fujian Province, China.

出版信息

Kaohsiung J Med Sci. 2020 Aug;36(8):581-591. doi: 10.1002/kjm2.12212. Epub 2020 Apr 20.

Abstract

To find out the role of hsa-miR-570-3p targeting CD274 in triple negative breast cancer (TNBC) via PI3K/AKT/mTOR signaling pathway. Hsa-miR-570-3p and CD274 expressions in 175 TNBC patients were detected by qRT-PCR and immunohistochemistry respectively. The human TNBC cell lines (MDA-MB-468 and MDA-MB-231) were used to verify the targeting relationship between hsa-miR-570-3p and CD274 via dual-luciferase reporter gene assay. Then, MDA-MB-468 and MDA-MB-231 cells were divided into Blank, miR-NC, miR-570-3p mimics, NC siRNA, CD274 siRNA, and miR-570-3p inhibitors + CD274 siRNA groups. Next, the biological activities of cells were detected by MTT, Cell-Light EdU, Annexin-V-FITC/PI, wound healing and Transwell invasion assays. Western blotting was conducted to detect protein expressions.MiR-570-3p expression was lower in tumor tissues than that in adjacent normal tissues, which was more obvious in CD274-positive TNBC patients, which targeted CD274 in TNBC cell lines. MiR-570-3p inhibited cell proliferation, invasion and migration, but induced cell apoptosis accompanying the upregulation of apoptotic proteins and downregulation of anti-apoptotic protein. CD274 siRNA had the similar results of miR-570-3p mimics, which could be reversed by miR-570-3p inhibitors. Besides, both miR-570-3p mimics and CD274 siRNA blocked PI3K/AKT/mTOR signaling pathway in TNBC cell lines. Hsa-miR-570-3p was downregulated and CD274 was upregulated in TNBC patients. Besides, hsa-miR-570-3p targeted CD274 to inhibit cell proliferation, invasion, migration, and induce cell apoptosis, which may be related to the suppression of PI3K/AKT/mTOR pathway.

摘要

为了通过 PI3K/AKT/mTOR 信号通路来发现 hsa-miR-570-3p 靶向 CD274 在三阴性乳腺癌(TNBC)中的作用。通过 qRT-PCR 和免疫组织化学分别检测了 175 例 TNBC 患者中 hsa-miR-570-3p 和 CD274 的表达。使用人 TNBC 细胞系(MDA-MB-468 和 MDA-MB-231)通过双荧光素酶报告基因检测验证 hsa-miR-570-3p 与 CD274 之间的靶向关系。然后,将 MDA-MB-468 和 MDA-MB-231 细胞分为空白组、miR-NC 组、miR-570-3p 模拟物组、NC siRNA 组、CD274 siRNA 组和 miR-570-3p 抑制剂+CD274 siRNA 组。接下来,通过 MTT、Cell-Light EdU、Annexin-V-FITC/PI、伤口愈合和 Transwell 侵袭实验检测细胞的生物学活性。通过 Western blot 检测蛋白表达。肿瘤组织中 miR-570-3p 的表达低于相邻正常组织,在 CD274 阳性的 TNBC 患者中更为明显,在 TNBC 细胞系中靶向 CD274。miR-570-3p 抑制细胞增殖、侵袭和迁移,但诱导细胞凋亡,同时上调凋亡蛋白和下调抗凋亡蛋白。CD274 siRNA 具有与 miR-570-3p 模拟物相似的结果,miR-570-3p 抑制剂可逆转其作用。此外,miR-570-3p 模拟物和 CD274 siRNA 均可阻断 TNBC 细胞系中的 PI3K/AKT/mTOR 信号通路。TNBC 患者中 hsa-miR-570-3p 下调,CD274 上调。此外,hsa-miR-570-3p 靶向 CD274 抑制细胞增殖、侵袭和迁移,并诱导细胞凋亡,这可能与抑制 PI3K/AKT/mTOR 通路有关。

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