Effect of Annexin A1 gene on the proliferation and invasion of esophageal squamous cell carcinoma cells and its regulatory mechanisms.
Effect of Annexin A1 gene on the proliferation and invasion of esophageal squamous cell carcinoma cells and its regulatory mechanisms.
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DOI：
10.3892/ijmm.2016.2840
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发表时间：
2017-02
影响因子：
5.4
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Han G;Lu K;Huang J;Ye J;Dai S;Ye Y;Zhang L
本研究旨在探讨膜联蛋白A1（Annexin A1，ANXA 1）对食管鳞癌（esophageal squamous cell carcinoma，ESCC）细胞增殖、迁移和侵袭的影响及其可能的作用机制。在构建ANXA 1过表达质粒后，我们将该质粒和/或microRNA（miRNA）-196a模拟物转染到ESCC细胞（Eca 109细胞系）中。采用四甲基偶氮唑蓝（MTT）比色法和Transwell小室法分别检测细胞增殖、迁移和侵袭能力。Western blot检测ANXA 1、Snail和E-cadherin蛋白表达水平。RT-PCR检测miRNA-196 a的表达。我们的研究结果显示，ANXA 1表达上调的细胞转染ANXA 1过表达质粒，细胞增殖，迁移和侵袭显着增加（p=0.004，p<0.001和p=0.011，分别）。在用miRNA-196 a模拟物转染的细胞中，miRNA-196 a表达显著上调（p<0.001）。然而，在用ANXA 1过表达质粒转染的细胞中，miRNA-196 a表达下调。此外，在用miRNA-196 a模拟物转染的细胞中，细胞增殖、迁移和侵袭显著降低（分别为p=0.027、p=0.009和p=0.021）。在ANXA 1过表达质粒转染的细胞中，Snail的表达上调，E-cadherin的表达下调。然而，在用miRNA-196 a模拟物转染的细胞中观察到相反的情况。因此，我们的研究结果表明，ANXA 1促进Eca 109细胞的增殖，并增加Snail的表达，而它抑制E-cadherin的表达，从而增强ESCC细胞的迁移和侵袭。miRNA-196 a负调控ANXA 1的表达，从而抑制食管鳞癌细胞的增殖、侵袭和转移。
The aim of this study was to examine the effect of Annexin A1 (ANXA1) on the proliferation, migration and invasion of esophageal squamous cell carcinoma (ESCC) cells and its possible mechanisms of action. After constructing the ANXA1 overexpression plasmid, we transfected this plasmid and/or microRNA (miRNA)-196a mimic into ESCC cells (Eca109 cell line). Methyl thiazolyl tetrazolium (MTT) assay and Transwell chamber assay were performed to determine cell proliferation, migration and invasion, respectively. Western blot analysis was used to examine the protein expression levels of ANXA1, Snail and E-cadherin. RT-PCR was used to detect the expression of miRNA-196a. Our results revealed that ANXA1 expression was upregulated in the cells transfected with the ANXA1 overexpression plasmid, and cell proliferation, migration and invasion were significantly increased (p=0.004, p<0.001 and p=0.011, respectively). In the cells transfected with the miRNA-196a mimic, miRNA-196a expression was significantly upregulated (p<0.001). However, miRNA-196a expression was downregulated in the cells transfected with the ANXA1 overexpression plasmid. In addition, in the cells transfected with the miRNA-196a mimic, cell proliferation, migration and invasion were significantly decreased (p=0.027, p=0.009 and p=0.021, respectively). In the cells transfected with the ANXA1 overexpression plasmid, the expression of Snail was upregulated and that of E-cadherin was downregulated. However, the opposite was observed in the cells transfected with the miRNA-196a mimic. Our findings thus demonstrate that ANXA1 promotes the proliferation of Eca109 cells, and increases the expression of Snail, whereas it inhibits that of E-cadherin, thus enhancing the migration and invasion of ESCC cells. miRNA-196a negatively regulates the expression of ANXA1, thereby inhibiting the proliferation, invasion and metastasis of ESCC cells.