目的:探讨 IL-13对人肺腺癌细胞(A549)IL-13受体α2(IL-13Rα2)表达及增殖、迁移的影响。方法采用定量逆转录聚合酶链式反应(qRT-PCR)、Western blot 法检测 IL-13刺激 A549细胞后,检测 IL-13Rα2的转录和蛋白质表达水平;酶联免疫吸附实验(ELISA)检测细胞培养上清液中的可溶型 IL-13受体α2(sIL-13Rα2)含量;流式细胞术(FCM)检测胞膜型 IL-13受体α2(memIL-13Rα2)、胞内型 IL-13受体 Rα2(iIL-13Rα2)的细胞数。MTT 法、Transwell 实验和细胞划痕实验观察 IL-13对 A549细 胞 增 殖 及 迁 移 作 用 的 影 响。结果 A549细 胞 可 表 达IL-13Rα2,并且 IL-13在较低质量浓度(10、20 ng·mL-1)时能上调 A549细胞 IL-13Rα2的表达水平,而对增殖和迁移无显著影响;在较高质量浓度(50、100 ng·mL-1)时对 IL-13Rα2的上调作用不明显,但能促进细胞增殖和迁移。另外低质量浓度 IL-13(10、20 ng·mL-1)对 IL-13Rα2整体水平、sIL-13Rα2、memIL-13Rα2以及 iIL-13Rα2的表达均有不同程度的上调(P <0.05);高质量浓度 IL-13(50、100 ng·mL-1)刺激条件下 IL-13Rα2整体水平的上调不明显且不再有剂量依赖性,sIL-13Rα2水平无明显改变、memIL-13Rα2表达略有增加、iIL-13Rα2水平明显下调(P <0.05)。结论IL-13对人肺腺癌 A549细胞 IL-13Rα2表达水平及增殖、迁移的影响具有双相性和差异性;IL-13Rα2在肺腺癌 A549中发挥抑制 IL-13促细胞增殖和迁移的作用。
Objective To investigate the effects of interleukin 13(IL-13)on IL-13 receptor α2 (IL-13Rα2),proliferation and migration in human lung adenocarcinoma A549 cells.Methods The expression of IL-13Rα2 mRNA and protein in A549 cells was determined after IL-13 stimulation by qRT-PCR and Western blot,respectively.The content of soluble IL-13Rα2(sIL-13Rα2)in cell culture supernatant was measured by ELISA.The number of cell expressing membrane IL-13Rα2 (memIL-13Rα2)and intracellular IL-13Rα2 (iIL-13Rα2)was detected by flow cytometry.The effects of IL-13 on proliferation and migration of A549 cells were evaluated by MTT,transwell assay and scratch test.Results IL-13Rα2 was expressed in A549 cells.IL-13 treatment at lower concentrations(10 and 20 ng · mL-1 )significantly increased the expression of IL-13Rα2,sIL-13Rα2,memIL-13Rα2 and intracellular IL-13Rα2(iIL-13Rα2),but had no effects on the prolifera-tion and migration of A549 cells.However,the higher concentrations of IL-13(50 and 100 ng· mL-1 )did not change the expression of IL-13Rα2 and sIL-13Rα2,slightly increased the expres-sion of memIL-13Rα2,and significantly decreased the expression of iIL-13Rα2.Furthermore,the higher concentrations of IL-13 significantly promoted the proliferation and migration of A549 cells.Conclusion There were “bipolarity”and “difference”in IL-13Rα2 expression,proliferation and migration in human lung adenocarcinoma A549 cells after IL-13 stimulation.IL-13Rα2 can in-hibit the effects of IL-13 on the proliferation and migration of A549 cells.