虫草素对人肝癌细胞迁移及侵袭的机制研究

Mechanism of cordycepin inhibiting migration and invasion of human hepatoma cells

摘要:

目的 研究虫草素对人肝癌细胞Bel-7402迁移和侵袭的影响，并探讨其作用机制。方法 以人肝癌细胞Bel-7402为研究对象，虫草素处理Bel-7402细胞。MTT法检测细胞活力；划痕实验检测细胞迁移能力；侵袭小室实验检测细胞侵袭能力；实时荧光定量聚合酶链反应（qRT-PCR）检测MMP-2和MMP-9 mRNA 表达情况；Western blotting实验检测MMP-2、MMP-9、ERK、p-ERK、EGFR及p-EGFR蛋白表达水平。结果 虫草素在0～80?μmol/L时对细胞活力无明显影响；而在80～640?μmol/L时，细胞活力出现下降；在320和640?μmol/L时显著降低（P?<0.05）。40和80?μmol/L虫草素均能显著抑制细胞迁移和侵袭活动（P?< 0.05）。与空白对照组比较，虫草素（40和80?μmol/L）处理Bel-7402细胞48?h后，MMP-2和MMP-9 mRNA 及蛋白表达均下降（P?<0.05）。与空白对照组比较，虫草素（40和80?μmol/L）处理明显下调p-ERK和p-EGFR蛋白表达（P?<0.05）。结论 虫草素可能通过EGFR-ERK通路介导MMP-2和MMP-9表达下调，从而调控人肝癌细胞Bel-7402的迁移和侵袭活动。

Abstract:

Objective To study the effect of cordycepin (CY) on the migration and invasion of hepatoma cell line Bel-7402, and to explore its mechanism. Methods Bel-7402 cells were used as the research model. Bel-7402 cells were treated with CY. Cell viability was detected by MTT assay, cell migration ability was detected by wound healing assay, and cell invasion ability was studied with invasion chamber assay. MMP-2 and MMP-9 mRNA expression were detected by qRT-PCR. The expression levels of MMP-2, MMP-9, ERK, p-ERK, EGFR and p-EGFR were detected by Western blotting. Results CY had no significant effect on cell viability at 0 to 80?μmol/L. Cell viability decreased at 80 to 640?μmol/L and significantly decreased at 320 and 640 μmol/L (P?