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[膜联蛋白A8通过EGFR-AKT-mTOR信号通路调控人非小细胞肺癌细胞A549的增殖]

[ANXA8 Regulates Proliferation of Human Non-Small Lung Cancer Cells A549 via EGFR-AKT-mTOR Signaling Pathway].

作者信息

Zhou G-Z, Sun Y-H, Shi Y-Y, Zhang Q, Zhang L, Cui L-Q, Sun G-C

机构信息

College of Bioengineering, Henan University of Technology, Zhengzhou, 450001 China.

Henan Academy of Fishery Sciences, Zhengzhou, 450044 China.

出版信息

Mol Biol (Mosk). 2021 Sep-Oct;55(5):870-880. doi: 10.31857/S0026898421050116.

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.

摘要

膜联蛋白A8(ANXA8)是膜联蛋白家族的成员,据报道它可调节多种癌症细胞过程,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不清楚。我们之前的转录组研究表明,在姜黄素类似物(MHMD)处理的人非小肺癌细胞(A549细胞系)中,ANXA8 mRNA表达下调。在此,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,并与正常人支气管上皮细胞(BE-AS-2B细胞系)进行比较。通过转染pEGFP-ANXA8重组载体过表达ANXA8促进了A549细胞的增殖和迁移。此外,在转染ANXA8的A549细胞中,细胞周期蛋白cyclin E1上调。使用RNA干扰技术敲低ANXA8可降低A549细胞活力并抑制其体外迁移。在A549细胞中,通过敲低或过表达ANXA8也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4EBP1,这证实了ANXA8对EGFR/Akt/mTOR信号通路的激活作用。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支持。

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