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紫杉醇对乳腺癌干细胞中某些微小RNA(miRNA)谱的疗效研究。

Investigation of the efficacy of paclitaxel on some miRNAs profiles in breast cancer stem cells.

作者信息

Ertürk Elif, Ari Ferda, Akgün Oğuzhan, Ulukaya Engin, Küçükali Cem İsmail, Zeybek Ümit

机构信息

Vocational School of Health Services, Bursa Uludağ University, Bursa Turkey.

Department of Biology, Science and Art Faculty, Bursa Uludağ University, Bursa Turkey.

出版信息

Turk J Biol. 2021 Oct 18;45(5):613-623. doi: 10.3906/biy-2103-46. eCollection 2021.

Abstract

Understanding of the functions of microRNAs in breast cancer and breast cancer stem cells have been a hope for the development of new molecular targeted therapies. Here, it is aimed to investigate the differences in the expression levels of let-7a, miR-10b, miR-21, miR-125b, miR-145, miR-155, miR-200c, miR-221, miR-222 and miR-335, which associated with gene and proteins in MCF-7 (parental) and MCF-7s (Mammosphere/stem cell-enriched population/CD44+/CD24-cells) cells treated with paclitaxel. MCF-7s were obtained from parental MCF-7 cells. Cytotoxic activity of paclitaxel was determined by ATP assay. Total RNA isolation and cDNA conversion were performed from the samples. Changes in expression levels of miRNAs were examined by RT-qPCR. Identified target genes and proteins of miRNAs were analyzed with RT-qPCR and western blot analysis, respectively. miR-125b was significantly expressed (2.0946-fold; p = 0.021) in MCF-7s cells compared to control after treatment with paclitaxel. Downregulation of SMO, STAT3, NANOG, OCT4, SOX2, ERBB2 and ERBB3 and upregulation of TP53 genes were significant after 48 h treatment in MCF-7s cells. Protein expressions of SOX2, OCT4, SMAD4, SOX2 and OCT4 also decreased. Paclitaxel induces miR-125b expression in MCF-7s cells. Upregulation of miR-125b may be used as a biomarker for the prediction of response to paclitaxel treatment in breast cancer.

摘要

了解微小RNA在乳腺癌和乳腺癌干细胞中的功能一直是开发新的分子靶向疗法的希望所在。本文旨在研究与基因和蛋白质相关的let-7a、miR-10b、miR-21、miR-125b、miR-145、miR-155、miR-200c、miR-221、miR-222和miR-335在紫杉醇处理的MCF-7(亲本)和MCF-7s(乳腺球/富含干细胞群体/CD44+/CD24-细胞)细胞中的表达水平差异。MCF-7s细胞取自亲本MCF-7细胞。通过ATP检测法测定紫杉醇的细胞毒性活性。从样品中进行总RNA分离和cDNA转化。通过RT-qPCR检测微小RNA表达水平的变化。分别用RT-qPCR和蛋白质印迹分析对已鉴定的微小RNA的靶基因和蛋白质进行分析。与紫杉醇处理后的对照组相比,miR-125b在MCF-7s细胞中显著表达(2.0946倍;p = 0.021)。在MCF-7s细胞中处理48小时后,SMO、STAT3、NANOG、OCT4、SOX2、ERBB2和ERBB3基因下调以及TP53基因上调均具有显著性。SOX2、OCT4、SMAD4、SOX2和OCT4的蛋白质表达也降低。紫杉醇诱导MCF-7s细胞中miR-125b表达。miR-125b的上调可作为预测乳腺癌对紫杉醇治疗反应的生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1edd/8574192/24ca38eeb08c/turkjbio-45-613-fig001.jpg

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