Abstract

Annexin A8 (ANXA8) is a member of the annexin family, which had been reported to regulate multiple cancer cellular processes including proliferation, metastasis and inflammation. However, the specific role of ANXA8 in lung cancer cell biology remains unknown. Our previous transcriptome study revealed that ANXA8 mRNA was downregulated in curcumin analog (MHMD)-treated human non-small lung cancer cells (A549 cell line). Here, we continued to study the ANXA8 expression in A549 cells using reverse transcription-quantitative PCR and Western blotting, compared with that in human normal bronchial epithelium cells (BE-AS-2B cell line). Overexpression of ANXA8 via transfection of pEGFP-ANXA8 recombinant vector contributed to the proliferation and migration of A549 cells. Moreover, the cell cycle protein cyclin E1 was upregulated in ANXA8-transfected A549 cells. Knockdown of ANXA8 using an RNA interference technique decreased A549 cell viability and restrained their migration in vitro. The expression levels of multiple cellular factors, including EGFR, PI3K, Akt, mTOR, p70S6K and 4EBP1, in the epidermal growth factor receptor (EGFR) signaling pathway were also altered by ANXA8 knockdown or overexpression in A549 cells, which confirmed the activation of the EGFR/Akt/mTOR signaling pathway by ANXA8. The present results provided evidence to support further investigation of the functional identification of ANXA8 in lung cancer cells in the future.

中文翻译：

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ANXA8 通过 EGFR-AKT-mTOR 信号通路调节人类非小肺癌细胞 A549 的增殖

摘要

膜联蛋白 A8 (ANXA8) 是膜联蛋白家族的成员，据报道其可调节多种癌细胞过程，包括增殖、转移和炎症。然而，ANXA8 在肺癌细胞生物学中的具体作用仍然未知。我们之前的转录组研究表明，ANXA8 mRNA 在姜黄素类似物 (MHMD) 处理的人类非小肺癌细胞（A549 细胞系）中下调。在这里，我们继续使用逆转录-定量 PCR 和蛋白质印迹法研究 A549 细胞中 ANXA8 的表达，并与人正常支气管上皮细胞（BE-AS-2B 细胞系）进行比较。ANXA8 的过度表达通过转染 pEGFP-ANXA8 重组载体有助于 A549 细胞的增殖和迁移。此外，细胞周期蛋白细胞周期蛋白 E1 在 ANXA8 转染的 A549 细胞中上调。使用 RNA 干扰技术敲除 ANXA8会降低 A549 细胞的活力并抑制它们的体外迁移。ANXA8还改变了表皮生长因子受体 (EGFR) 信号通路中多种细胞因子的表达水平，包括 EGFR、PI3K、Akt、mTOR、p70S6K 和 4EBP1A549 细胞中的敲低或过表达，证实了 ANXA8 激活了 EGFR/Akt/mTOR 信号通路。目前的结果为未来进一步研究ANXA8在肺癌细胞中的功能鉴定提供了证据。