| MiR-30a-5p通过泛素水解酶22抑制人宫颈癌细胞上皮-间质转化功能 |
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| 引用本文： | 王丽萍,何婧,许晓燕,陈小梅,李红芳. MiR-30a-5p通过泛素水解酶22抑制人宫颈癌细胞上皮-间质转化功能[J]. 中山大学学报(医学科学版), 2019, 40(5): 698 |
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| 作者姓名： | 王丽萍 何婧 许晓燕 陈小梅 李红芳 |
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| 作者单位： | 兰州市第一人民医院妇产科，甘肃兰州730050 |
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| 基金项目： | 甘肃省重点研发计划——社会发展类项目（17yf1fa141） |
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| 摘 要： | 【目的】研究microRNA-30a-5p（miR-30a-5p）对人宫颈癌Hela细胞上皮-间质转化功能的影响及其相关机制。【方法】宫颈癌Hela细胞株分别转染目的mir的模拟物和阴性对照模拟物，分别以30a-5p组、NC组命名并标记细胞。同时，以未经过处理的Hela细胞作为对照（Control组）。分别用逆转录-聚合酶链反应法检测各组宫颈癌细胞的miR-30a-5p含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Western-blot法检测3组细胞神经-钙粘素（N-cadherin）、α-连环蛋白（α-Catenin）和泛素水解酶22（USP22）表达水平。运用生物信息学方法预测miR-30a-5p的靶基因。采用Western blot法检测USP22过表达对miR-30a-5p抑制EMT的拮抗作用。双荧光素酶实验检测miR-30a-5p与USP22的关系。建立皮下移植瘤模型观察miR-30a-5p的体内作用。【结果】30a-5p组宫颈癌细胞miR-30a-5p的表达水平明显上调，表达水平为Control组的853.82（862.26~843.11）倍（P＜0.01）。30a-5p组侵袭细胞数量8.17（8.32~8.03）明显低于Control组（P＜0.01）。30a-5p组细胞N-cadherin蛋白的细胞内含量明显下降，α-Catenin蛋白的细胞内含量明显上升，USP22蛋白表达量明显降低。合并USP22过表达处理的30a-5p组宫颈癌细胞中N-cadherin蛋白表达量明显升高，α-Catenin蛋白表达量明显降低。双荧光素酶检验结果显示USP22为miR-30a-5p的下游靶基因（P＜0.01）。30a-5p组皮下移植瘤明显小于Control组（P＜0.01）。与Control组肿瘤组织相比，30a-5p组肿瘤组织miR-30a-5p的相对含量升高，USP22蛋白含量降低，N-cadherin蛋白的含量降低，α-Catenin蛋白含量升高。【结论】miR-30a-5p在宫颈癌Hela细胞中，可能通过靶向识别下游靶基因USP22，进而抑制其翻译。最终实现对宫颈癌细胞EMT过程的抑制。
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| 关 键 词： | 宫颈癌 微小RNA-30a-5p USP22 上皮-间质转化 |
| 收稿时间： | 2019-05-21 |
MiR-30a-5p Inhibits Epithelial Mesenchymal Transition Function in Human Cervical Cancer Cells by Targeting Ubiquitin-specific Protease 22 |
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| WANG Li-ping,HE Jing,XU Xiao-yan,CHEN Xiao-mei,LI Hong-fang. MiR-30a-5p Inhibits Epithelial Mesenchymal Transition Function in Human Cervical Cancer Cells by Targeting Ubiquitin-specific Protease 22[J]. Journal of Sun Yatsen University(Medical Sciences), 2019, 40(5): 698 |
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| Authors: | WANG Li-ping HE Jing XU Xiao-yan CHEN Xiao-mei LI Hong-fang |
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| Affiliation: | Department of Obstetrics & Gynecology，The First People′s Hospital of Lanzhou City，Lanzhou 730050，China |
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| Abstract: | 【Objective】To investigate the inhibitory effect and mechanism of microRNA- 30a-5p（miR-30a-5p）on epithelial mesenchymal transition in cervical cancer Hela cells.【Methods】Hela cervical cancer cell lines were transfected with miR-30a-5p mimics or negative control mimic，respectively，as 30a-5p or NC group. Control group was established with untreated Hela cervical cancer cells. miR- 30a-5p content in each group was detected by RT-PCR assay. Transwell assay was used to detect the invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of N- cadherin，α- Catenin and ubiquitin specific processing peptidase 22（USP22）protein in the 3 groups. Prediction target genes of miR-30a-5p by bioinformatics methods. Antagonistic effect of USP22 over-expression on miR-30a-5p inhibition of EMT was detected by western blot assay. The relationship between miR- 30a-5p and USP22 was detected by dual luciferase assay. Subcutaneous transplantation tumor model established，and the effect of miR- 30a- 5p in vivo was observed.【Results】The miR-30a-5p intracellular quantity in 30a-5p group Hela cells was up-regulated，and the expression level of miR- 30a-5p was 853.82（862.26~843.11）times higher than that of Control group（P＜0.01）. The number of invasive cells in 30a- 5p group was 8.17（8.32~8.03），which was significantly lower than that of Control group 62.33（63.52~60.19）（P＜0.01）. USP22 may be the target gene of miR-30a-5p. In 30a-5p group，the intracellular quantity of N- cadherin protein was decreased，the intracellular quantity of α- Catenin protein was increased，and the intracellular quantity of USP22 protein was decreased. The intracellular quantity of N-cadherin protein in 30a-5p group was down-regulated，the intracellular quantity of α-Catenin protein was increased，and the intracellular quantity of USP22 protein was reduced. After USP22 over-expression，the intracellular quantity of N-cadherin protein in cervical cancer cells of 30a-5p group was up-regulated，and the intracellular quantity of α-Catenin protein were down-regulated. Dual luciferase assay showed that USP22 is a downstream target gene of miR-30a-5p（P＜0.01）. Subcutaneous transplantation tumors in 30a-5p group were significantly smaller than those in Control group.【Conclusion】miR-30a-5p may inhibit the expression of EMT related protein through the downstream target gene USP22，and the epithelial mesenchymal transition function of cervical cancer Hela cells. |
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| Keywords: | cervical cancer microRNA- 30a-5p ubiquitin-specific protease 22 epithelial mesenchymal transition |
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