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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。
Objective To investigate the effect of YKL-40 on the migration of SKOV-3 ovarian cancer cells in vitro. Methods The expression of YKL-40 mRNA in ovarian cancer SKOV-3 cells was detected by RT-PCR. The expression of YKL-40 mRNA in SKOV-3 cells was detected by overexpression of YKL-40, qRT-PCR, ELISA and Western blotting Overexpression; Transw ell assay was used to detect the effect of YKL-40 overexpression on the migration ability of SKOV-3 cells; specific inhibitor was used to inhibit the p38 MAPK pathway in YKL-40 over-expressing SKOV-3 cells . Results The expression of YKL-40 mRNA was not detected in SKOV-3 cells. LV-SKOV-3 over-expressing YKL-40 cells was successfully constructed, and the infection rate was over 90%. Compared with control SKOV-3 Compared with the NC-SKOV-3 cells, the expression of YKL-40 in LV-SKOV-3 cells was significantly up-regulated both in mRNA and protein levels; Transwell cell migration assay showed that YKL-40 overexpressing LV The migration ability of SKOV-3 cells in NC-SKOV-3 cells was significantly enhanced compared with control cells. However, the migration ability of LV-SKOV-3 cells was significantly down-regulated after p38 MAPK pathway was inhibited by SB203580. Conclusion YKL-40 can significantly promote the migration of ovarian cancer SKOV-3 cells, which may be mediated by the p38 M APK pathway.