Table1_miR-92a-3p Promoted EMT via Targeting LATS1 in Cervical Cancer Stem Cells.XLS

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.

miR-92a-3p（microRNA-92a-3p，微小RNA-92a-3p）已被报道在多种癌症中存在表达失调，因此被认为是一种癌症相关微小RNA。然而，miR-92a-3p对宫颈癌（cervical cancer，CC）细胞生物学行为的影响目前仍不明确。本研究采用实时定量聚合酶链反应（Quantitative real-time PCR）检测宫颈癌干细胞中miR-92a-3p的表达水平；通过细胞计数试剂盒-8（Cell Counting Kit-8，CCK8）检测法、Transwell细胞侵袭实验及流式细胞术，分析miR-92a-3p与大肿瘤抑制因子1（large tumor suppressor 1，LATS1）对细胞增殖、侵袭及细胞周期转换的调控作用；利用荧光素酶报告基因实验验证miR-92a-3p与LATS1之间的靶向结合关系；采用蛋白质印迹法（Western Blotting）检测相关信号通路及蛋白的表达情况。癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据集分析结果显示，miR-92a-3p在宫颈癌组织中呈高表达，且与患者总生存期密切相关。miR-92a-3p可促进宫颈癌干细胞的增殖、侵袭及细胞周期转换。荧光素酶报告基因实验结果证实，miR-92a-3p可结合至LATS1启动子的3'非翻译区（3′-untranslated region，3′-UTR）。LATS1则可抑制宫颈癌干细胞的增殖、侵袭及细胞周期转换。蛋白质印迹实验结果显示，LATS1可下调PDZ结合基序转录共激活因子（transcriptional co-activator with PDZ-binding motif，TAZ）、波形蛋白（vimentin）及细胞周期蛋白E（cyclin E）的表达，同时上调E-钙粘蛋白（E-cadherin）的表达。重新表达LATS1可部分逆转miR-92a-3p对宫颈癌干细胞增殖、侵袭及细胞周期转换的调控作用，同时可恢复TAZ、E-钙粘蛋白、波形蛋白及细胞周期蛋白E的表达水平。miR-92a-3p通过靶向调控LATS1，进而调节TAZ与E-钙粘蛋白的表达，最终促进宫颈癌干细胞的恶性生物学行为。