YKL-40对卵巢癌SKOV-3细胞迁移能力的影响

发布时间：2018-03-24 09:01

本文选题：卵巢癌　切入点：YKL-　出处：《山东大学学报(医学版)》2017年01期

【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;Transw ell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 M APK通路介导的。
[Abstract]:Objective to investigate the effect of YKL-40 on the migration of ovarian cancer SKOV-3 cells in vitro. Methods RT-PCR was used to detect the expression of YKL-40 mRNA in ovarian cancer SKOV-3 cells. Overexpression of YKL-40qRT-PCRRT-ELISA-Western blotting in SKOV-3 cells was used to detect the effect of YKL-40 overexpression on the migration ability of SKOV-3 cells by Transw ell assay, and specific inhibitors inhibited p38 MAPK pathway in YKL-40 overexpression SKOV-3 cells. Results the expression of YKL-40 mRNA was not detected in SKOV-3 cells, and the YKL-40 overexpression cell line LV-SKOV-3 was successfully constructed, and the infection rate was over 90%. Compared with the empty vector lentivirus infected control SKOV-3 cell line NC-SKOV-3, the YKL-40 of LV-SKOV-3 cells expressed either mRNA or protein (total protein). Secretory protein) up-regulated the migration of Transwell cells. The results showed that the migration ability of LV-SKOV-3 cells which overexpressed YKL-40 was significantly higher than that of empty vector cells, and the migration ability of LV-SKOV-3 cells was significantly higher than that of empty vector cells. The migration ability of LV-SKOV-3 cells was down-regulated after p38 MAPK pathway p38 protein phosphorylation was inhibited by SB203580. Conclusion YKL-40 can significantly promote the migration of ovarian cancer SKOV-3 cells, which may be mediated by p38M APK pathway.
【作者单位】：山东大学齐鲁医院化疗科;
【基金】：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)
【分类号】：R737.31

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