[中性粒细胞胞外陷阱通过上调MMP9表达激活局灶黏附激酶,促进小鼠结直肠癌细胞的增殖和迁移]。

Yi He, Songlin Hou, Changyuan Memg
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引用次数: 0

摘要

目的探讨中性粒细胞胞外陷阱(NETs)对小鼠MC38结直肠癌细胞增殖和迁移的影响及其机制。方法提取小鼠脾中性粒细胞,体外离子霉素刺激后收集NETs。与MC38细胞共孵育后,采用CCK-8法检测MC38细胞的增殖能力,TranswellTM法和细胞划痕法检测MC38细胞的迁移能力。实时荧光定量PCR检测细胞基质金属蛋白酶2 (MMP2)和MMP9 mRNA的表达,Western blot检测MMP2、MMP9和局灶黏附激酶(FAK)、磷酸化FAK蛋白的表达。利用小干扰RNA (small interfering RNA, siRNA)沉默MMP9后,我们通过之前的方法检测NETs对MC38细胞增殖和迁移能力的影响以及相关分子表达的改变。结果NETs促进MC38细胞增殖和迁移,上调MMP9表达和FAK磷酸化水平。沉默MMP9抑制NETs促进MC38增殖和迁移,抑制FAK磷酸化。结论NETs上调MC38细胞MMP9表达,激活FAK信号通路,促进肿瘤细胞增殖和迁移。
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[Neutrophil extracellular traps activates focal adhesion kinase by upregulating MMP9 expression to promote proliferation and migration of mouse colorectal cancer cells].

Objective To investigate how the neutrophil extracellular traps (NETs) affect the proliferation and migration of mouse MC38 colorectal cancer cells and its mechanism. Methods Spleen neutrophils were extracted in mouse, followed by collection of NETs after ionomycin stimulation in vitro. The proliferation of MC38 cell was detected by CCK-8 assay, and migration ability were detected by TranswellTM and cell scratch assay, after co-incubation with MC38 cells. The mRNA expression of cellular matrix metalloproteinase 2 (MMP2) and MMP9 were detected by real-time fluorescence quantitative PCR, and the expression of MMP2, MMP9 and focal adhesion kinase (FAK), phosphorylated FAK protein were detected by Western blot. After silencing MMP9 using small interfering RNA (siRNA), the effect of NETs on the proliferation and migration ability of MC38 cells and the altered expression of related molecules were examined by previous approach. Results NETs promoted the proliferation and migration of MC38 cells and up-regulated the MMP9 expression and FAK phosphorylation. Silencing MMP9 inhibited the promotion of MC38 proliferation and migration by NETs and suppressed FAK phosphorylation. Conclusion NETs up-regulates MMP9 expression in MC38 cells, activates FAK signaling pathway and promotes tumor cell proliferation and migration.

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