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目的体外研究CXCL1对乳腺癌细胞增殖和迁移侵袭能力的影响及其作用机制。方法选取人乳腺癌细胞进行分组处理:空白对照组、CXCL1组、anti-CXCR2中和抗体组,用CCK8、克隆形成实验、流式凋亡、细胞划痕及Transwell迁移实验分别对各组细胞增殖活性、凋亡率、迁移侵袭能力进行检测,并用ELISA法对各组细胞内MMP-9蛋白、磷酸化AKT及活化NF-κB水平进行检测。结果 CCK8与克隆形成实验结果显示,与空白对照组相比CXCL1处理组细胞增殖活性提高,细胞克隆形成率增加(P<0.05),与空白对照组相比anti-CXCR2处理组细胞增殖活性降低,细胞克隆形成率降低(P<0.05);流式凋亡检测结果显示,与空白对照组相比CXCL1处理组细胞凋亡率降低(P<0.05),与空白对照组相比anti-CXCR2处理组细胞凋亡率增高(P<0.05);细胞划痕、Transwell迁移实验及细胞内MMP-9蛋白表达水平表明,与空白对照组相比CXCL1处理组细胞迁移侵袭能力增强(P<0.05),与空白对照组相比anti-CXCR2处理组细胞迁移侵袭能力减弱(P<0.05);细胞内p AKT/NF-κB蛋白检测表明,与空白对照组相比CXCL1处理组细胞内AKT磷酸化水平及NF-κB活化水平升高(P<0.05),与空白对照组相比anti-CXCR2处理组细胞内AKT磷酸化水平及NF-κB活化水平降低(P<0.05)。结论 CXCL1通过与CXCR2结合促进乳腺癌细胞增殖和迁移侵袭,其机制可能是激活了AKT/NF-κB信号转导通路。
Objective To investigate the effect of CXCL1 on the proliferation, migration and invasion of breast cancer cells in vitro and its mechanism. Methods The human breast cancer cells were selected and divided into groups: blank control group, CXCL1 group and anti-CXCR2 neutralizing antibody group. CCK8, colony formation assay, flow cytometry, cell scratch assay and Transwell migration assay were used to detect the cell proliferation Activity, apoptosis rate, migration and invasion ability were measured. The levels of MMP-9, phospho-AKT and NF-κB in each group were detected by ELISA. Results The results of CCK8 and colony formation assay showed that compared with the blank control group, the proliferation activity of CXCL1 group increased and the colony formation rate increased (P <0.05). Compared with the blank control group, the antiproliferative activity of anti-CXCR2 group decreased, The results of flow cytometry showed that the apoptosis rate of CXCL1-treated group was significantly lower than that of the blank control group (P <0.05). Compared with the blank control group, the anti-CXCR2-treated group (P <0.05). Scratches, Transwell migration assay and MMP-9 protein expression showed that compared with the control group, the migration and invasion ability of CXCL1-treated cells were enhanced (P <0.05) Compared with the blank control group, the cell migration and invasion ability of anti-CXCR2-treated group was weakened (P <0.05). The intracellular AKT / NF-κB protein assay showed that AKT phosphorylation and NF (P <0.05). Compared with the blank control group, the level of AKT phosphorylation and the activation of NF-κB in anti-CXCR2-treated cells decreased (P <0.05). Conclusion CXCL1 may promote the proliferation and migration of breast cancer cells by binding with CXCR2. The mechanism may be that AKT / NF-κB signal transduction pathway is activated.