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Characteristics of the microbiome in lung adenocarcinoma tissue from patients in Kunming city of southwestern China

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Abstract

The purpose of this study is to identify the characteristics of microbial communities in the lung cancer tissues from patients in Kunming sity of southwestern China and to compare the microbial differences at different clinical stages of lung cancer to uncover potential microbial biomarkers. In total, 40 tissue samples of primary lung adenocarcinoma were collected and further performed by 16S rRNA gene sequencing. The subjects were grouped according to TNM stages (T and N group), clinical stage, and smoke status, and the microbial differences in each group were compared. Analysis of sequence data to determine beta diversity, the UniFrac distance was calculated by QIIME and visualized by principal coordinate analysis (PCoA) using R (version 2.15.3). Microbiome abundance and diversity between different groups were calculated by t test or Wilcoxon rank sum test and drawn by R. The linear discriminant analysis effect size (LEfSe) method was utilized to compare relative abundances of all bacterial taxa between groups. A total of 951 OTUs were identified in the cancer tissues. No significant difference has been found in the alpha diversity within all the groups. Beta diversity significantly differed in the N, T, and clinical stage groups. By LEfSe analysis, eight differential taxa including Bifidobacterium were identified in the N group. In the T1 and T2 group, the LEfSe result identified five phyla and ten genera. The differential genera were Moraxella, Dolosigranulum, unidentified_Corynebacteriaceae, and Citrobacter in the T2 group and Bifidobacterium, Alistipes, Akkermansia, Blautia, Lactobacillus, as well as Faecalibaculum in the T1 group. Differential bacterial composition and abundance were also observed in the clinical stage group. This study confirmed that by 16S rRNA sequencing, we identified the dominant microbe of lung cancer tissue in different groups. Bifidobacterium may play an essential role in lymph node metastasis and tumor progression, providing a specific potential microbial biomarker for lung adenocarcinoma. PCR products were subject to vertical electrophoresis on 2% agarose gels, and a colloid recovery kit (Qiagen, Valencia, CA) was applied to recover the target bands. Libraries were generated by the TruSeq DNA PCR-Free Sample Preparation Kit (Illumina, San Diego, USA), and the concentrations were quantitated with a Qubit fluorometer. Finally, the qualified libraries were sequenced by NovaSeq6000 (Illumina).

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Acknowledgements

We sincerely thank Hangzhou Lianhe Medical Laboratory Co., LTD, for providing the help in 16S rRNA sequencing and results interpretation.

Funding

This study was supported by the Yunnan Provincial Department of Science and Technology-Kunming Medical University Applied Basic Research Project [No.202001AY070001-025].

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Author notes

  1. Manjun Chen and Yue Cui contributed equally to this study.

Authors and Affiliations

  1. Department of Thoracic Surgery, The First Affiliated Hospital of Kunming Medical University, No.295 Xichang Road, Yunnan, 650032, Kunming, China

    Manjun Chen, Yue Cui, Chao Liu, Xinghe Tong, Mengtian Wang, Yunping Zhao & Xiaobo Chen

  2. Department of Medicine, Dinfectome Inc., Nanjing, Jiangsu, 210032, China

    Chunman Wu & Yong Liu

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  1. Manjun Chen
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Contributions

Sinuo Song and Xiaobo Chen designed the study and drafted the manuscript. Yunping Zhao and Yanan Bao collected the patients’ samples and clinical information. Xuan He and Duozhi Shi performed the 16S rRNA sequencing. Yue Cui and Xingming Zhu analyzed the OTU data. All the authors read and approved the final manuscript.

Corresponding author

Correspondence to Xiaobo Chen.

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Ethical approval

The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). The study was approved by the Medical Ethics Committee of The First Affiliated Hospital of Kunming Medical University Kunming(Code:HKMU20180532). All the subjects have signed informed consent for inclusion before participating in the study.

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All the authors have approved the manuscript for publication.

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The authors declare no competing interests.

Preprint statement

A preprint has been published earlier [https://doi.org/10.21203/rs.3.rs-46069/v1].

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Chen, M., Cui, Y., Liu, C. et al. Characteristics of the microbiome in lung adenocarcinoma tissue from patients in Kunming city of southwestern China. Environ Sci Pollut Res 30, 49992–50001 (2023). https://doi.org/10.1007/s11356-023-25528-1

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