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miR-92a-3p通过靶向LATS1促进宫颈癌干细胞中的上皮-间质转化

miR-92a-3p Promoted EMT Targeting LATS1 in Cervical Cancer Stem Cells.

作者信息

Liu Shuangyue, Chu Liping, Xie Mingzhu, Ma Lisha, An Hongmei, Zhang Wen, Deng Jihong

机构信息

Department of Gynecology, Kunming Maternity and Child Care Hospital, Kunming, China.

出版信息

Front Cell Dev Biol. 2021 Nov 18;9:757747. doi: 10.3389/fcell.2021.757747. eCollection 2021.

Abstract

miR-92a-3p (microRNA-92a-3p) has been reported to be dysregulated in several cancers, and as such, it is considered to be a cancer-related microRNA. However, the influence of miR-92a-3p on biological behaviors in cervical cancer (CC) still remains unclear. Quantitative real-time PCR was used to detect miR-92a-3p levels in CC stem cells. Here, Cell Counting Kit-8 (CCK8) assay, Transwell cell invasion assay and flow cytometry assay were used to characterize the effects that miR-92a-3p and large tumor suppressor l (LATS1) had on proliferation, invasion and cell cycle transition. The luciferase reporter gene assay was used to verify the targeting relationship between miR-92a-3p and LATS1. Western Blotting was used to investigate the related signaling pathways and proteins. Data from The Cancer Genome Atlas (TCGA) showed that miR-92a-3p was upregulated in CC tissues and closely associated with overall survival. miR-92a-3p promoted proliferation, invasion and cell cycle transition in CC stem cells. The luciferase reporter assay showed that miR-92a-3p bound to the 3'-untranslated region (3'-UTR) of the LATS1 promoter. LATS1 inhibited proliferation, invasion and cell cycle transition. Results measured by Western Blotting showed that LATS1 downregulated expressions of transcriptional co-activator with PDZ-binding motif (TAZ), vimentin and cyclin E, but upregulated the expression of E-cadherin. Re-expression of LATS1 partly reversed the effects of miR-92a-3p on proliferation, invasion and cell cycle transition, as well as on TAZ, E-cadherin, vimentin, and cyclin E. miR-92a-3p promoted the malignant behavior of CC stem cells by targeting LATS1, which regulated TAZ and E-cadherin.

摘要

据报道,miR-92a-3p(微小RNA-92a-3p)在多种癌症中表达失调,因此被认为是一种与癌症相关的微小RNA。然而,miR-92a-3p对宫颈癌(CC)生物学行为的影响仍不清楚。采用定量实时聚合酶链反应检测CC干细胞中miR-92a-3p的水平。在此,使用细胞计数试剂盒-8(CCK8)检测、Transwell细胞侵袭检测和流式细胞术检测来表征miR-92a-3p和大肿瘤抑制因子1(LATS1)对增殖、侵袭和细胞周期转换的影响。荧光素酶报告基因检测用于验证miR-92a-3p与LATS1之间的靶向关系。蛋白质免疫印迹法用于研究相关信号通路和蛋白质。来自癌症基因组图谱(TCGA)的数据显示,miR-92a-3p在CC组织中上调,且与总生存期密切相关。miR-92a-3p促进CC干细胞的增殖、侵袭和细胞周期转换。荧光素酶报告基因检测表明,miR-92a-3p与LATS1启动子的3'-非翻译区(3'-UTR)结合。LATS1抑制增殖、侵袭和细胞周期转换。蛋白质免疫印迹法检测结果显示,LATS1下调了含PDZ结合基序的转录共激活因子(TAZ)、波形蛋白和细胞周期蛋白E的表达,但上调了E-钙黏蛋白的表达。LATS1的重新表达部分逆转了miR-92a-3p对增殖、侵袭和细胞周期转换以及对TAZ、E-钙黏蛋白、波形蛋白和细胞周期蛋白E的影响。miR-92a-3p通过靶向LATS1促进CC干细胞的恶性行为,而LATS1调节TAZ和E-钙黏蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c24a/8639224/c9ad511eb239/fcell-09-757747-g001.jpg

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