摘要：
目的 探讨在肺腺癌细胞株A549 中,晚期糖基化终末产物受体(RAGE)及其胞内信号分子DIAPH1 的表达及对细胞迁移及凋亡能力的影响.方法 ① 以肺腺癌细胞株 A549 及正常人支气管上皮细胞株BEAS-2B 为研究对象,利用qRT-PCR、Western blot 检测RAGE 及DIAPH1 在二者中的表达;② 以10、100 μg /ml RAGE 配体CML-AGE 及1、10、100 μg /ml RAGE 配体S100B 处理A549 细胞,划痕实验检测其对细胞迁移能力的影响;③ 以25、50、100 μg /ml RAGE 配体 CML-AGE 处理A549 细胞,qRT-PCR 法检测凋亡相关基因 BCL-2 和BAX 的表达情况.结果 ① qRT-PCR、Western blot 检测结果显示A549 细胞中RAGE 及DIAPH1 表达较 BEAS-2B 细胞明显下调(P < 0. 001);② 10、100 μg /ml RAGE 配体CML-AGE 及1、10、100 μg /ml RAGE 配体S100B处理对A549 细胞的迁移能力皆有明显抑制作用,且作用呈浓度依赖性(P < 0. 01),差异有统计学意义;③ 以25、50、 100 μg /ml RAGE 配体CML-AGE 处理A549 细胞后,抗凋亡基因BCL-2 表达下调,促凋亡基因BAX 表达上调,其作用呈浓度依赖性(P < 0. 05),差异有统计学意义.结论 RAGE及DIAPH1 在肺腺癌细胞株A549 中的表达低于正常人支气管上皮细胞BEAS-2B.RAGE 配体在一定程度上可以抑制 A549 细胞的迁移,促进其凋亡,有望成为肺腺癌治疗新的靶点.%Objective To explore the expression of the receptor for advanced glycosylation end products(RAGE) and its intracellular signaling molecules DIAPH1 in lung adenocarcinoma A549 cells and the effect of RAGE ligands on cell migration and apoptosis. Methods The expressions of RAGE and DIAPH1 in lung adenocarcinoma A549 cells and human bronchial epithelial cells BEAS-2B were tested by qRT-PCR and Western blot. A549 cells was treated with 10,100 μg /ml RAGE ligands CML-AGE and 1,10,100 μg /ml S100B,and wound healing test was used to identify the effect of migration ability. A549 cells was treated with 25,50,100 μg /ml RAGE ligands CMLAGE, the gene expression of BCL-2 and BAX were tested by using qRT-PCR. Results The results of qRT-PCR and Western blot showed,compared with human bronchial epithelium cells BEAS-2B,the expression of RAGE and DIAPH1 were both significantly down-regulated in lung adenocarcinoma A549 cells (P < 0. 001). After treated with 10,100 μg /ml RAGE ligands CML-AGE and 1,10,100 μg /ml S100B ,both groups showed the ligands inhibit lung adenocarcinoma A549 cells migration in concentration-depend manners (P < 0. 01). After treated with 25, 50,100 μg /ml RAGE ligands CML-AGE,the expression of anti-apoptotic gene BCL-2 was down-regulated and proapoptotic gene BAX was upregulated in the experimental group in concentration-depend manners(P < 0. 05),the difference was significant. Conclusion The expression levels of RAGE and DIAPH1 in lung adenocarcinoma A549 cells are both significantly lower than human bronchial epithelium cells BEAS-2B. RAGE ligands can inhibit cells migration and promote cell apoptosis in lung adenocarcinoma A549 cells and may provide a new target for the therapy of lung adenocarcinoma cells.
