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山东大学学报(医学版) ›› 2017, Vol. 55 ›› Issue (1): 33-38.doi: 10.6040/j.issn.1671-7554.0.2016.1304

• 基础医学 • 上一篇    下一篇

YKL-40对卵巢癌SKOV-3细胞迁移能力的影响

陆衡,刘延国,李曙光,陈晓康,田琦,衣翠华,王秀问   

  1. 山东大学齐鲁医院化疗科, 山东 济南 250012
  • 收稿日期:2016-10-12 出版日期:2017-01-10 发布日期:2017-01-10
  • 通讯作者: 王秀问. E-mail: xiuwenwang12@sdu.edu.cn E-mail:xiuwenwang12@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(81372530,81502615);山东大学齐鲁医院临床研究项目(2014QLKY30)

Regulation of epithelial ovarian carcinoma SKOV-3 cell migration by YKL-40

LU Heng, LIU Yanguo, LI Shuguang, CHEN Xiaokang, TIAN Qi, YI Cuihua, WANG Xiuwen   

  1. Department of Medical Oncology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2016-10-12 Online:2017-01-10 Published:2017-01-10

摘要: 目的 探讨YKL-40对卵巢癌SKOV-3细胞体外迁移能力的影响。 方法 采用RT-PCR检测卵巢癌SKOV-3细胞YKL-40 mRNA水平的表达;采用慢病毒感染的方法在SKOV-3细胞过表达YKL-40,qRT-PCR、ELISA、Western blotting验证YKL-40的过表达;Transwell实验检测YKL-40过表达对SKOV-3细胞迁移能力的影响;特异性抑制剂抑制YKL-40过表达SKOV-3细胞中的p38 MAPK通路,检测其对细胞迁移能力的影响。 结果 SKOV-3细胞中未检测到YKL-40 mRNA的表达;成功构建了YKL-40过表达细胞株LV-SKOV-3,感染率达90%以上;与空载体慢病毒感染的对照SKOV-3细胞(NC-SKOV-3)相比,LV-SKOV-3细胞YKL-40无论mRNA表达还是蛋白水平(细胞总蛋白、分泌蛋白)均明显上调;Transwell细胞迁移实验表明过表达YKL-40的LV-SKOV-3细胞较空载体对照细胞NC-SKOV-3迁移能力显著增强,而通过SB203580抑制p38 MAPK通路p38蛋白磷酸化后,LV-SKOV-3细胞的迁移能力明显下调。 结论 YKL-40可显著促进卵巢癌SKOV-3细胞的迁移,这一作用可能是通过p38 MAPK通路介导的。

关键词: 卵巢癌, 细胞迁移, p38 MAPK通路, YKL-40

Abstract: Objective To investigate the effect of YKL-40 on epithelial ovarian cancer SKOV-3 cell migration in vitro and to explore the underlying mechanism. Methods YKL-40 mRNA expression in SKOV-3 cells was detected by RT-PCR. SKOV-3 cells were transfected with modified lentiviral vectors containing YKL-40 gene and YKL-40 overexpression was validated by qRT-PCR, ELISA and Western blotting. Transwell assay was used to evaluate the effect of YKL-40 on SKOV-3 cell migration. p38MAPK pathway was repressed by specific inhibitors and its effect on YKL-40 overexpressed SKOV-3 cell migration was further investigated. Results YKL-40 mRNA was not detected in SKOV-3 cells. After lentivirus transfection and puromycin selection, YKL-40 overexpressed cell line LV-SKOV-3 and negative control cell line NC-SKOV-3 were established. Further analyses confirmed YKL-40 was significantly increased in LV-SKOV-3 cells on both mRNA and protein levels compared to NC-SKOV-3. After YKL-40 overexpression, the migration ability of LV-SKOV-3 was enhanced while LV-SKOV-3 cell migration was decreased after inhibition of p38MAPK pathway via SB203580. Conclusion YKL-40 significantly promotes the migration ability of SKOV-3 cells, which is partly mediated by p38MAPK pathway.

Key words: Cell migration, p38MAPK pathway, Ovarian cancer, YKL-40

中图分类号: 

  • R737.31
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