解剖学报

解剖学报 ›› 2022, Vol. 53 ›› Issue (1): 42-49.doi: 10.16098/j.issn.0529-1356.2022.01.006

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MiR-515-5p通过调控硫酸软骨素蛋白聚糖4表达对卵巢癌A2780细胞增殖和转移的影响及其机制

江红1 黄艳丽1 邢辉1 汪黎明2 郭红1*
  

  1. 1.湖北文理学院附属医院襄阳市中心医院妇产科，湖北 襄阳 441021;  2.华中科技大学同济医学院湖北省妇幼保健院妇产科，  武汉 430070
  • 收稿日期:2020-08-18 修回日期:2020-11-24 出版日期:2022-02-06 发布日期:2022-02-06
  • 通讯作者: 郭红 E-mail:ghnmnno@163.com
  • 基金资助:
    湖北省自然科学基金支持

Effects of miR-515-5p on the proliferation and metastasis of ovarian cancer cell A2780 by regulating the expression of chondroitin sulfate proteoglycan 4 and its mechanism

JIANG Hong1  HUANG Yan-li1  XING Hui1  WANG Li-ming2  GUO Hong1*   

  1. 1.Department of Gynecology and Obstetrics，Xiangyang Central Hospital，Affiliated Hospital of Hubei University of Arts and Science，Hubei Xiangyang 441021, China;   2. Department of Gynecology and Obstetrics, Hubei Provincial Maternal and Child Health Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430070, China
  • Received:2020-08-18 Revised:2020-11-24 Online:2022-02-06 Published:2022-02-06
  • Contact: GUO Hong E-mail:ghnmnno@163.com

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摘要：

目的  探讨miR-515-5p对硫酸软骨素蛋白聚糖4（CSPG4）的靶向调控作用，以及对卵巢癌细胞系A2780细胞增殖和转移的影响。   方法  通过生物信息学工具预测miR-515-5p的靶基因。Real-time PCR和Western blotting法检测65例卵巢癌组织和与其对应的癌旁组织中miR-515-5p和CSPG4的表达。将A2780细胞分为miR-NC组、miR-515-5p组、si-NC组、si-CSPG4组、miR-515-5p+pcDNA组和miR-515-5p+pcDNA-CSPG4组。MTT法检测细胞增殖活力，Transwell检测细胞迁移和侵袭数，Western blotting检测细胞周期蛋白D1（cyclinD1）、P21、基质金属蛋白酶（MMP）-2和MMP-9蛋白的表达。双荧光素酶报告基因实验和Western blotting验证miR-515-5p对CSPG4基因的调控作用。   结果  生物信息学分析显示，CSPG4是miR-515-5p的潜在靶基因之一。与癌旁组织相比，卵巢癌组织miR-515-5p的表达较低，CSPG4的表达较高（P<0.05）。与miR-NC组、miR-515-5p组相比，A2780细胞cyclinD1、MMP-2和MMP-9蛋白的表达较低，P21蛋白的表达较高，细胞活力较低，细胞迁移和侵袭数较少（P<0.05）。与si-NC组相比，si-CSPG4组A2780细胞cyclinD1、MMP-2和MMP-9蛋白的表达较低，P21蛋白的表达较高，细胞活力较低，细胞迁移和侵袭数较少（P<0.05）。与miR-515-5p+pcDNA组相比，miR-515-5p+pcDNA-CSPG4组A2780细胞cyclinD1、MMP-2和MMP-9蛋白的表达较高，P21蛋白的表达较低，细胞活力较高，细胞迁移和侵袭数较多（P<0.05）。MiR-515-5p靶向并负性调控CSPG4表达。   结论  MiR-515-5p通过靶向CSPG4可抑制卵巢癌细胞A2780增殖和转移。

关键词: 卵巢癌, 微小RAN-515-5p, 硫酸软骨素蛋白聚糖4, 细胞增殖, 转移, Transwell实验, 人 

Abstract:

Objective  To investigate the targeted regulation of miR-515-5p on chondroitin sulfate proteoglycan 4 (CSPG4) and its effect on the proliferation and metastasis of ovarian cancer cell A2780.    Methods  The target genes of miR-515-5p were predicted by bioinformatics tools. Real-time PCR and Western blotting were used to detect the expression of miR-515-5p and CSPG4 in 65 cases ovarian cancer tissues and adjacent tissues. Ovarian cancer cells A2780 were divided into miR-NC group, miR-515-5p group, si-NC group, si-CSPG4 group, miR-515-5p + pcDNA group, miR-515-5p + pcDNA-CSPG4 group. MTT assay was used to detect cell proliferation. Transell was applied to detect cell migration and invasion, and Western blotting was selected to detect cyclinD1, P21, matrix metalloproteinase(MMP)-2 and MMP-9 protein expression. The double luciferase reporter gene experiment and Western blotting confirmed the regulation effect of miR-515-5p on CSPG4 gene.    Results  Bioinformatics analysis showed that CSPG4 was one of the potential target genes of miR-515-5p. Compared with the adjacent tissues, the expression of miR-515-5p was lower in ovarian cancer tissues, and the expression of CSPG4 was higher (P<0.05). Compared with the miR-NC group, the expression of cyclinD1, MMP-2 and MMP-9 protein was lower in A2780 cells of miR-515-5p group, the expression of P21 protein was higher, the cell viability was lower, and the number of cell migration and invasion was lower (P<0.05). Compared with the si-NC group, the expression of cyclinD1, MMP-2 and MMP-9 proteins were lower in A2780 cells in si-CSPG4 group, P21 protein was higher, cell viability was lower, and the number of cell migration and invasion was lower (P<0.05). Compared with the miR-515-5p + pcDNA group, the expression of cyclinD1, MMP-2 and MMP-9 proteins was higher in A2780 cells of miR-515-5p + pcDNA-CSPG4 group, the expression of P21 protein was lower, and the cell viability was higher, the number of cell   igration and invasion was higher (P<0.05). MiR-515-5p targeted and negatively regulated CSPG4 expression.    Conclusion  MiR-515-5p could inhibit the proliferation and metastasis of ovarian cancer cell A2780 by targeting CSPG4. 

Key words: Ovarian cancer, MicroRNA-515-5p, Chondroitin sucfac proteoglycan 4, Cell proliferation, Metastasis, Transwell assays, Human

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