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长链非编码RNA ZFAS1/微小RNA-589在乳腺癌细胞增殖、侵袭和迁移过程中调控PTEN/PI3K/AKT信号通路。

LncRNA ZFAS1/miR-589 regulates the PTEN/PI3K/AKT signal pathway in the proliferation, invasion and migration of breast cancer cells.

作者信息

Zhang Song, Wang Jin, Yao Tingjing, Tao Min

机构信息

Department of Medical Oncology, First Affiliated Hospital of Soochow University, Suzhou, 215006, Jiangsu, China.

Department of Surgical Oncology, First Affiliated Hospital of Bengbu Medical College, Bengbu, 233004, Anhui, China.

出版信息

Cytotechnology. 2020 Jun;72(3):415-425. doi: 10.1007/s10616-020-00388-6. Epub 2020 Mar 18.

Abstract

Breast cancer (BC) is a common clinical disease and the second leading cause of cancer death in women. Long noncoding RNA (lncRNA) and microRNA (miRNA) are reported to be involved in the development of BC. The present study aimed to investigate whether LncRNA ZFAS1 could regulate the proliferation, invasion and migration of breast cancer cells by targeting miR-589 through PTEN/PI3K/AKT signal pathway. The expression of ZFAS1 and miR-589 in BC cells and transfection effects were determined by RT-qPCR analysis. The abilities of proliferation, colony formation, invasion and migration of breast cancer cells were analyzed by CCK-8 assay, colony formation assay, transwell assay and wound healing assay respectively. The expression of MMP2, MMP9, Bcl2, Bax, cleaved caspase3, PTEN, p-PI3K, p-AKT, PI3K and AKT was detected by Western blot. The flow cytometry analysis was used to detect cell apoptosis. As a result, ZFAS1 expression was increased and miR-589 expression was decreased in BC cells. And, miR-589 was demonstrated to be a target of ZFAS1. ZFAS1 overexpression could inhibit the proliferation, colony formation, invasion and migration of BC cells while miR-589 overexpression could reverse the changes. In addition, ZFAS1 overexpression suppressed the expression of PI3K/AKT signal pathway by activating the PTEN expression while miR-589 overexpression could reverse the changes. Moreover, PTEN is one of the gene targets of miR-589. In conclusion, this study indicated that ZFAS1 inhibited the proliferation, invasion and migration of breast cancer cells by targeting miR-589 through regulating the PTEN/PI3K/AKT signal pathway.

摘要

乳腺癌(BC)是一种常见的临床疾病,是女性癌症死亡的第二大主要原因。据报道,长链非编码RNA(lncRNA)和微小RNA(miRNA)参与了乳腺癌的发生发展。本研究旨在探讨lncRNA ZFAS1是否能通过PTEN/PI3K/AKT信号通路靶向miR-589来调节乳腺癌细胞的增殖、侵袭和迁移。通过RT-qPCR分析测定BC细胞中ZFAS1和miR-589的表达及转染效果。分别采用CCK-8法、集落形成试验、Transwell试验和伤口愈合试验分析乳腺癌细胞的增殖、集落形成、侵袭和迁移能力。通过蛋白质免疫印迹法检测MMP2、MMP9、Bcl2、Bax、裂解的caspase3、PTEN、p-PI3K、p-AKT、PI3K和AKT的表达。采用流式细胞术分析检测细胞凋亡。结果显示,BC细胞中ZFAS1表达升高,miR-589表达降低。并且,miR-589被证明是ZFAS1的靶标。ZFAS1过表达可抑制BC细胞的增殖、集落形成、侵袭和迁移,而miR-589过表达可逆转这些变化。此外,ZFAS1过表达通过激活PTEN表达抑制PI3K/AKT信号通路的表达,而miR-589过表达可逆转这些变化。而且,PTEN是miR-589的基因靶标之一。总之,本研究表明ZFAS1通过调节PTEN/PI3K/AKT信号通路靶向miR-589抑制乳腺癌细胞的增殖、侵袭和迁移。

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