Background: ANXA2 plays a very important role in cancer progression. chemokine ligand 18 (CCL18) is associated with the invasion, migration, metastasis and poor prognosis of lung adenocarcinoma (LUAD). In this study, we aimed to explore whether CCL18 promotes LUAD invasion through ANXA2, and its role and molecular mechanism in LUAD invasion.
Methods: Western blot was used to detect ANXA2 expression in LUAD tissues and adjacent non-tumor tissues, the transfection efficiency of SiANXA2#2 in cells and the role of ANXA2 as an upstream regulator in the AKT/cofilin signaling pathway. In vitro cytological experiments such as chemotaxis experiment and transwell invasion test was used to explore the mechanism of ANXA2 on LUAD metastasis. F-actin polymerization experiment and Western blot were used to detect whether invasion ability alteration of SiANXA2#2 A549 cells are related to F-actin.
Results: Western blot analysis showed that compared with adjacent non-tumor tissues, the protein expression level of ANXA2 in cancer tissues increased (P<0.05). In the chemotaxis experiment and invasion experiment, the chemotaxis and invasion ability induced by CCL18 decreased when ANXA2 knockdowned (P<0.05). Compared with the control group, F-actin polymerization was significantly lower in ANXA2 knockdown group, while phosphorylation of AKT at Ser473 and Thr308 and phosphorylation of Cofilin and LIMK were reduced in ANXA2 knockdown group (P<0.05).
Conclusions: ANXA2 knockdown can reduce the invasive effect of CCL18 on LUAD cells by reducing phosphorylation of AKT and downstream pathways.
【中文题目：CCL18通过ANXA2促进肺腺癌的侵袭】 【中文摘要：背景与目的 ANXA2在癌症进展中起着非常重要的作用，趋化因子18（chemokine ligand 18, CCL18）与肺腺癌（lung adenocarcinoma, LUAD）的侵袭、迁移、转移及预后不良有关。本研究旨在探究CCL18是否通过ANXA2促进LUAD侵袭以及其在LUAD侵袭中的作用和分子机制。方法 Western blot检测LUAD组织与癌周正常组织中ANXA2表达量，并检测转染效率及ANXA2作为上游调节剂在AKT/cofilin信号通路中的作用。细胞趋化实验、Transwell侵袭实验等实验探讨ANXA2对LUAD的作用机理。F-actin聚合实验和Western blot检测转染SiRNA的A549细胞侵袭是否与F-actin相关。结果 与相邻非肿瘤组织相比，癌组织中ANXA2的蛋白表达水平升高（P<0.05）。敲低ANXA2组的LUAD细胞中CCL18诱导的趋化运动能力和侵袭能力下降（P<0.05）。与对照组相比，敲低ANXA2的LUAD细胞的F-actin聚合明显降低，而敲低ANXA2的LUAD细胞中AKT在Ser473和Thr308位点的磷酸化和cofilin、LIMK的磷酸化水平均降低（P<0.05）。结论 敲低ANXA2可以通过降低AKT及下游通路磷酸化，进而降低CCL18对LUAD细胞的侵袭性的影响。】 【中文关键词：ANXA2；CCL18；肺肿瘤；侵袭】.
Keywords: ANXA2; CCL18; Invasion; Lung neoplasms.