[摘要] 目的: 探讨微小RNA（miR）-760 在人宫颈鳞状细胞癌（CSCC）组织和细胞中的表达及其对SiHa 细胞增殖、凋亡、侵袭、迁移、EMT影响的分子机制。方法: 选用2015 年4 月至2018 年8 月山东省聊城市妇幼保健院妇科手术切除、经病理确诊的80 例CSCC组织及相应的癌旁组织标本和40 例子宫肌瘤切除术获取的正常宫颈组织标本，应用qPCR检测CSCC组织、癌旁组织和正常宫颈组织、人CSCC细胞系SiHa、HCC94 和人宫颈鳞状上皮永生化细胞株H8 中miR-760 的表达水平，分析miR-760 表达与CSCC患者临床病理特征的相关性。用脂质体转染法，将miR-760 mimics 和NC-mimics 质粒分别转染至SiHa 细胞，用qPCR检测SiHa 细胞中miR-760 的表达，用CCK-8 实验和流式细胞术分别检测SiHa 细胞增殖能力和凋亡水平，用Transwell 实验检测SiHa 细胞的侵袭和迁移能力，WB检测SiHa 细胞EMT相关蛋白上皮型钙黏蛋白（E-cadherin）、波形蛋白（vimentin）和神经型钙黏蛋白（N-cadherin）的表达变化。用生物学信息法预测FOXA1 与miR-760 的靶向关系，用双荧光素酶报告基因验证miR-760 对FOXA1的直接靶向调控作用。结果：CSCC组织中miR-760 表达明显低于癌旁组织及正常宫颈组织（均P<0.01），miR-760 表达与患者淋巴结转移和临床分期密切相关（均P<0.01）。SiHa、HCC94 细胞中miR-760 的表达明显低于H8 细胞（均P<0.01）。通过转染miR-760 mimics 上调miR-760 表达，能够显著抑制SiHa 细胞的增殖能力和侵袭、迁移能力（均P<0.01）、促进其凋亡（P<0.01）,并上调Ecadherin的表达（P<0.01）、下调vimentin 和N-cadherin 的表达（均P<0.01）。FOXA1 是miR-760 的直接靶基因（P<0.01），上调miR-760 表达显著抑制SiHa 细胞中FOXA1 mRNA和蛋白的表达（均P<0.05）。结论：在CSCC组织和细胞中miR-760 低表达，其通过靶向作用FOXA1 基因调控SiHa细胞的增殖、侵袭、迁移并促进凋亡，同时影响癌细胞的EMT进程。

[Abstract] Objective: To investigate the expression of microRNA (miR) -760 in human cervical squamous cell carcinoma (CSCC) tissues and cells, and it’s effects on the proliferation, apoptosis, invasion, migration and epithelial-mesenchymal transition (EMT) of SiHa cells, as well as its molecular mechanism. Methods: Eighty pairs of CSCC cancerous and corresponding para-cancerous tissue specimens which were pathologically confirmed and 40 cases of normal cervical tissue specimens obtained by myomectomy in Liaocheng Maternal and Child Health Hospital of Shandong Province from April 2015 to August 2018 were selected. The expression of miR-760 in CSCC tissues, para-cancerous tissues and normal cervical tissues, human CSCC cell lines (SiHa, HCC94) and human cervical squamous epithelial immortalized H8 cells were detected by qPCR, and the relationship between miR-760 and clinicopathological characteristics of CSCC patients was analyzed. miR-760 mimics and NC-mimics plasmids were transfected into SiHa cells by liposome transfection.The expression of miR-760 in SiHa cells was detected by qPCR, the proliferation activity and apoptosis rate were detected by CCK-8 test and flow cytometry, respectively. The invasion and migration of SiHa cells were detected by Transwell assay. The expres-sions of EMT-related proteins, such as E-cadherin, vimentin and N-cadherin, in SiHa cells were detected by WB. Bioinformatics was used to predict the targeting relationship between FOXA1 and miR-760, and double luciferase reporter gene assay was used to verify the direct regulation of miR-760 on FOXA1. Results: The expression of miR-760 in CSCC tissues was significantly lower than that in para-cancerous tissues and normal cervical tissues (all P<0.01), and the expression of miR-760 was closely related to lymphnode metastasis and clinical stage (all P<0.01). The expression of miR-760 in SiHa and HCC94 cells was significantly lower than that in H8 cells (all P<0.01). Up-regulation of the expression of miR-760 could significantly inhibit the proliferation, invasion and migration of SiHa cells (all P<0.01), promote apoptosis (P<0.01), up-regulate the expression of E-cadherin protein (P<0.01), and down-regulate the protein expressions of vimentin and N-cadherin (all P<0.01).FOXA1 was a direct target gene of miR-760 (P<0.01). Up-regulation of miR-760 significantly inhibited the mRNA and protein expressions of FOXA1 in SiHa cells (all P<0.05). Conclusion: The expression of miR-760 is down-regulated in CSCC tissues and cells, and it can regulate the proliferation, invasion, migration and apoptosis of CSCC cells by targeting FOXA1.

山东省医药卫生科技发展计划资助项目（No.2016WS0583）