宫颈鳞状细胞癌是女性生殖系统中最常见的癌症类型。本研究旨在探究microRNA-9-5p（miR-9-5p）对宫颈鳞状细胞的迁移、侵袭和上皮-间充质转化（EMT）过程的影响。利用生物信息学预测了与E-cadherin（E-cad）结合的miRNAs，利用癌症基因组计划（TCGA）数据库分析和提取了部分宫颈鳞状细胞癌组织和正常宫颈组织中显著差异表达的miRNAs，选定miR-9-5p作为主要研究对象。将野生型E-cad的转化区域（UTR）（E-cad-WT 3'-UTR）或突变型E-cad的3'-UTR（E-Cad-MUT 3'-UTR）与miR-9-5p mimic正常对照组（NC）共转染人类胚胎肾（293T）细胞。通过双荧光素酶检测miR-9-5p与E-cad的关系。 通过定量实时PCR检测miR-9-5p在正常宫颈上皮细胞系（H8）和宫颈鳞状细胞系（C33A，siha，caski和Me180）中的表达。然后，将实验分为以下组：阻断对照组、过表达对照组（miR-9-5p mimic组）、miR-9-5p过表达组（mimic组）、抑制表达对照组（inhibitor-NC组）和miR-9-5p抑制表达组（inhibitor组）。通过划痕实验检测迁移能力的变化。采用Transwell侵袭实验分析侵袭能力的变化，并通过定量实时PCR和Western blot检测E-cad和Vimentin的mRNA和蛋白质变化。miR-9-5p与E-cad有定向结合关系。与正常宫颈组织H8细胞系相比，miR-9-5p在宫颈癌细胞系（C33A，siha，caski和Me180）中高表达（均P<0.05）。在miR-9-5p mimic细胞中，E-cad-MUT的荧光素酶活性升高，与E-cad-WT相比（P<0.05）。与空白对照组相比，miR-9-5p mimic组中的E-cad蛋白质和mRNA表达均下降（均P<0.05）；miR-9-5p抑制剂组中则上升（均P<0.05）。与H8细胞系相比，宫颈鳞状细胞系中miR-9-5p高表达（均P<0.05）。与mimic-NC组相比，caski和Me180细胞入侵到膜下的距离、伤口愈合距离以及Vimentin的mRNA和蛋白质表达在miR-9-5p mimic组中均上升（均P<0.05）；E-cad的mRNA和蛋白质表达则下降（均P<0.05）。与inhibitor-NC组相比，miR-9-5p抑制剂组中的伤口愈合距离、侵袭膜下的caski和Me180细胞数量以及Vimentin的mRNA和蛋白质表达均下降（均P<0.05），但E-cad的mRNA和蛋白质表达增加（均P<0.05）。miR-9-5p在宫颈鳞状细胞癌中高表达，可以增加癌细胞的迁移和侵袭能力，促进EMT过程。

Cervical squamous cell carcinoma is the most common cancer in female reproductive system. This study aims to explore the effect of microRNA-9-5p (miR-9-5p) on the migration, invasion, and epithelial-mesenchymal transition (EMT) process of cervical squamous cells.Bioinformatics were used to predict the miRNAs that could bind to E-cadherin (E-cad). The Cancer Genome Atlas (TCGA) database was used to analyze and extract significantly differentially expressed miRNAs from part of cervical squamous cell carcinoma tissues and normal cervical tissues, and miR-9-5p was selected as the main research target. The translated regions (UTR) of wild-type E-cad (E-cad-WT 3'-UTR) or the 3'-UTR of mutant E-cad (E-Cad-MUT 3'-UTR) was transfected with miR-9-5p mimic normal control (NC), and miR-9-5p mimic was co-transfected human embryonic kidney cells (293T). The relationship between miR-9-5p and E-cad was detected by double luciferase assay. The expression of miR-9-5p in normal cervical epithelial cell lines (H8) and cervical squamous cell lines (C33A, siha, caski and Me180) were detected by quantitative real-time PCR. Then, the experiments were divided into groups as follows: a block control group, an overexpression control group (mimic-NC group), a miR-95p overexpression group (mimic group), an inhibitory expression control group (inhibitor-NC group), and a miR-9-5p inhibitory expression group (inhibitor group). The changes of migration ability were detected by scratch assay. Transwell invasion assay was used to analyze the changes of invasion ability, and the mRNA and protein changes of E-cad and vimentin were detected by quantitative real-time PCR and Western blotting.MiR-9-5p had a targeting binding relationship with E-cad. Compared with the normal cervical tissue H8 cell line, the miR-9-5p was highly expressed in cervical cancer cell lines (C33A, siha, caski and Me180) (all P<0.05). The luciferase activity of E-cad-MUT was increased compared with that of E-cad-WT in miR-9-5p mimic cells (P<0.05). Compared with the blank control group, the protein and mRNA expressions of E-cad were decreased in the miR-9-5p mimic group (both P<0.05), which were increased in the miR-9-5p inhibitor group (both P<0.05). Compared with H8 cell line, the miR-9-5p was highly expressed in the cervical squamous cell lines (all P<0.05). Compared with the mimic-NC group, the distance of wound healing, the number of caski and Me180 cells invaded below the membrane, and the mRNA and protein expressions of vimentin were all increased in the miR-9-5p mimic group (all P<0.05), while the mRNA and protein of E-cad were decreased (both P<0.05). Compared with the inhibitor-NC group, the distance of wound healing, the number of caski and Me180 cells invading the membrane, and the mRNA and protein expressions of vimentin were decreased in the miR-9-5p inhibitor group (all P<0.05), but the mRNA and protein expressions of E-cad were increased (both P<0.05).The miR-9-5p is highly expressed in cervical squamous cell carcinoma, which can increase the migration and invasion ability, and promote the EMT process of cancer cells.