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Metformin inhibits human non-small cell lung cancer by regulating AMPK–CEBPB–PDL1 signaling pathway

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Abstract

Metformin has been found to have inhibitory effects on a variety of tumors. However, its effects on non-small cell lung cancer (NSCLC) remain unclear. We demonstrated that metformin could inhibit the proliferation of A549 and H1299 cells. RNA transcriptome sequencing revealed that PDL1 was significantly downregulated in both cell types following treatment with metformin (P < 0.001). Jaspar analysis and chromatin immunoprecipitation showed that CEBPB could directly bind the promoter region of PDL1. Western blotting showed that protein expression of the isoforms CEBPB-LAP*, CEBPB-LAP, and CEBPB-LIP was significantly upregulated and the LIP/LAP ratio was increased. Gene chip analysis showed that PDL1 was significantly upregulated in A549-CEBPB-LAP cells and significantly downregulated in A549-CEBPB-LIP cells (P < 0.05) compared with CEBPB-NC cells. Dual-luciferase reporter gene assay showed that CEBPB-LAP overexpression could promote transcription of PDL1 and CEBPB-LIP overexpression could inhibit the process. Functional assays showed that the changes in CEBPB isoforms affected the function of NSCLC cells. Western blotting showed that metformin could regulate the function of NSCLC cells via AMPK–CEBPB–PDL1 signaling. Animal experiments showed that tumor growth was significantly inhibited by metformin, and atezolizumab and metformin had a synergistic effect on tumor growth. A total of 1247 patients were retrospectively analyzed, including 166 and 1081 patients in metformin and control groups, respectively. The positive rate of PDL1 was lower than that of the control group (HR = 0.338, 95% CI = 0.235–0.487; P < 0.001). In conclusion, metformin inhibited the proliferation of NSCLC cells and played an anti-tumor role in an AMPK–CEBPB–PDL1 signaling-dependent manner.

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Abbreviations

AMPK:

AMP-activated protein kinase.

ChIP:

Chromatin immunoprecipitation

DEG:

Differential expressed genes

EMT:

Epithelial interstitial transformation

FC:

Fold change

HR:

Hazard ratio

NSCLC:

Non-small cell lung cancer

OE:

Overexpressed

PDL1:

Programmed cell death ligand-1

qPCR:

Quantitative polymerase chain reaction

SPF:

Specific pathogen-free

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Acknowledgements

This work was supported by grants from National Natural Science Foundation of China (No. 81401875) and the Research Program of Shanghai Health Commission (Grant No. 20204Y022). We have asked the International Science Editing Corporation for editing the language.

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  1. Tao Lu, Ming Li and Mengnan Zhao have contributed equally to this work.

Authors and Affiliations

  1. Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Tao Lu, Ming Li, Mengnan Zhao, Yiwei Huang, Guoshu Bi, Jiaqi Liang, Zhencong Chen, Yuansheng Zheng, Junjie Xi, Zongwu Lin, Cheng Zhan, Wei Jiang, Qun Wang & Lijie Tan

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  1. Tao Lu
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Contributions

CZ and WJ take responsibility for the integrity of the work as a whole. CZ, WJ, and TL conceived the study. TL, ML, and MNZ performed most of the bioinformatics analysis and wrote the manuscript. GSB, YWH, JQL, ZCC, and YSZ collected the tumor samples and analyzed the data of clinical characteristics. JJX, ZWL, and QW helped project design and manuscript editing. LLT supervised this study.

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Correspondence to Cheng Zhan or Wei Jiang.

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Lu, T., Li, M., Zhao, M. et al. Metformin inhibits human non-small cell lung cancer by regulating AMPK–CEBPB–PDL1 signaling pathway. Cancer Immunol Immunother 71, 1733–1746 (2022). https://doi.org/10.1007/s00262-021-03116-x

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