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长链非编码 RNA SPRY4-IT1 通过与 TCF7L2 竞争性结合 miR-6882-3p 调节乳腺癌细胞干性。

LncRNA SPRY4-IT1 regulates breast cancer cell stemness through competitively binding miR-6882-3p with TCF7L2.

机构信息

Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang, China.

Liaoning Key Laboratory of Molecular Targeted Anti-Tumor Drug Development and Evaluation, China Medical University, Shenyang, China.

出版信息

J Cell Mol Med. 2020 Jan;24(1):772-784. doi: 10.1111/jcmm.14786. Epub 2019 Nov 17.

Abstract

SPRY4-intronic transcript 1 has been found in several kinds of cancers, but the role of SPRY4-IT1 in breast cancer stem cells has not been studied. We investigated whether SPRY4-IT1 is involved in the promotion of breast cancer stem cells (BCSCs). We used qRT-PCR to detect the expression of SPRY4-IT1 in MCF-7 cells and MCF-7 cancer stem cells (MCF-7 CSCs). The effects of SPRY4-IT1 on the proliferation and renewal ability of breast cancer cells were investigated by in vitro and in vivo assays (ie in situ hybridization, colony formation assay, sphere formation assay, flow cytometry assay, western blotting, xenograft model and immunohistochemistry). The mechanism of SPPRY4-IT1 as a ceRNA was studied by a dual-luciferase reporter assay and bioinformatic analysis. In our study, SPRY4-IT1 was up-regulated in MCF-7 CSCs compared with MCF-7 cells, and high SPRY4-IT1 expression was related to reduced breast cancer patient survival. Furthermore, SPRY4-IT1 overexpression promoted breast cancer cell proliferation and stemness in vitro and in vivo. In addition, SPRY4-IT1 knockdown suppressed BCSC renewal ability and stemness maintenance in vivo and in vitro. The dual-luciferase reporter assays indicated that SPRY4-IT1 as a sponge for miR-6882-3p repressed transcription factor 7-like 2 (TCF7L2) expression. Taken together, these findings demonstrated that SPRY4-IT1 promotes proliferation and stemness of breast cancer cells as well as renewal ability and stemness maintenance of BCSCs by increasing the expression of TCF7L2 through targeting miR-6882-3p.

摘要

SPRY4 内含子转录本 1 已在多种癌症中被发现,但 SPRY4-IT1 在乳腺癌干细胞中的作用尚未被研究。我们研究了 SPRY4-IT1 是否参与促进乳腺癌干细胞(BCSCs)。我们使用 qRT-PCR 检测 MCF-7 细胞和 MCF-7 癌症干细胞(MCF-7 CSCs)中 SPRY4-IT1 的表达。通过体外和体内实验(原位杂交、集落形成实验、球体形成实验、流式细胞术分析、Western blot 分析、异种移植模型和免疫组织化学)研究 SPRY4-IT1 对乳腺癌细胞增殖和更新能力的影响。通过双荧光素酶报告基因检测和生物信息学分析研究 SPRY4-IT1 作为 ceRNA 的机制。在我们的研究中,与 MCF-7 细胞相比,SPRY4-IT1 在 MCF-7 CSCs 中上调,并且高 SPRY4-IT1 表达与降低的乳腺癌患者生存率相关。此外,SPRY4-IT1 过表达促进了乳腺癌细胞在体外和体内的增殖和干性。此外,SPRY4-IT1 敲低抑制了体内和体外 BCSC 更新能力和干性维持。双荧光素酶报告基因检测表明,SPRY4-IT1 作为 miR-6882-3p 的海绵通过靶向 miR-6882-3p 抑制转录因子 7 样 2(TCF7L2)的表达。总之,这些发现表明,SPRY4-IT1 通过靶向 miR-6882-3p 增加 TCF7L2 的表达,促进乳腺癌细胞的增殖和干性以及 BCSC 的更新能力和干性维持。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71b8/6933354/fce4bc9e3dda/JCMM-24-772-g001.jpg

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