缺氧对肺腺癌细胞甲酰肽受体及其侵袭力的影响

Effects of Hypoxia on FPR1 Expression and Invasiveness of A549 Cells

目的探讨缺氧对人肺腺癌细胞甲酰肽受体1(formyl peptide receptor 1,FPR1)表达及其对肿瘤细胞侵袭力的影响。方法将人肺腺癌细胞株A549暴露于常氧(21%O2)和缺氧(1%O2)条件下培养4、12、24 h,应用RT-PCR检测FPR1 mRNA水平,采用Western blot检测培养上清FPR1激动剂脂联素1(Annexin 1,ANX1)的表达量。用趋化实验检测缺氧对A549细胞FPR1活性的影响。用FPR1外源性激动剂fMLP(formyl-met-leu-phe)刺激细胞后,用RT-PCR观察FPR1 mRNA的变化;同时采用体外侵袭实验检测其对肿瘤细胞侵袭能力的影响。结果与常氧组相比,缺氧上调A549细胞FPR1表达的同时也增加A549细胞FPR1的趋化活性,增加FPR1内源性激动剂Annexin 1的产生。fMLP刺激细胞后,FPR1 mRNA表达水平上调。体外侵袭实验结果表明,缺氧细胞侵袭能力显著增强,加入FPR1拮抗剂Boc2后,缺氧细胞侵袭能力减弱(P<0.05)。结论缺氧增加肺腺癌细胞FPR1激动剂产生与FPR1基因表达。FPR1激动剂的产生在缺氧诱导的FPR1基因表达上调中发挥了作用。缺氧引起A549细胞侵袭力增强与FPR1有关。

Objective To evaluate the effects of hypoxia on the expression of formyl peptide receptor 1（FPR1） and the invasiveness of lung adenocarcinoma cell lines A549.Methods A549 cells were exposed to either normoxia（21%O2） or hypoxia（1%O2） condition for 4,12 and 24 h.The expressions of FPR1 mRNA levels were measured by RT-PCR.The expressions of Annexin 1 protein and FPR1 activity were investigated by Western blot and chemotaxis assay.Cells invasiveness was assayed by matrigel transwell.Results Compared with normoxia group,the FPR1 mRNA in hypoxia group increased;moreover,Annexin 1 in culture medium supernatants of A549 cells also functional increased.Treatment of FPR1 agonist fMLP increased the mRNA level of FPR1 in A549 cells.Hypoxia significantly increased the number of A549 cells invaded across the transwell matrixgel,and FPR specific antagonist Boc2 attenuated the hypoxia-induced invasiveness of A549 cells.Conclusion The results suggest that hypoxia plays an important role in the augmentation of the FPR1 expression and its agonist production,which were involved in the invasiveness of lung cancer cells.