Abstract
Purpose
Lung adenocarcinoma (LUAD) accounts for approximately half of patients in lung cancer. Cancer-associated fibroblasts (CAFs) are the major component in the tumor microenvironment (TME). Targeting CAFs is a promising therapeutic strategy for cancer treatment. However, therapeutic targets of CAFs in LUAD remains largely unclear.
Methods
Seven CAFs and nine normal fibroblasts (NFs) were isolated from tumor and paratumor tissues of LUAD patients undergoing surgery, respectively. RNA-seq and bioinformatics analysis were performed to identify the differentially expressed genes (DEGs) and their functions in CAFs compared with NFs. DEGs of ten overlaying were obtained from RNA-seq, our previously reported lncRNA microarray and public datasets (E-MTAB-6149, E-MTAB-6653) and validated by RT-qPCR. Nik-related kinase (NRK) was further validated by RT-qPCR, immunofluorescence (IF), Western Blot (WB) in vitro, and in Cancer Cell Line Encyclopedia (CCLE) database. Survival analysis was performed on Kaplan–Meier plotter.
Results
A total of 1799 DEGs were identified, including 650 upregulated DEGs and 1149 downregulated DEGs. The upregulated and downregulated DEGs were mostly enriched in extracellular matrix (ECM) functions and in glycolysis/gluconeogenesis pathways. Interestingly, NRK was the most significantly upregulated overlaying DEGs which was rarely associated with CAFs before. NRK was predominantly expressed in CAFs, but weakly expressed in NFs, normal lung bronchial epithelial cell line BEAS-2B, LUAD cell lines A549 and H1299, as well as in the majority of 191 lung cancer cell lines including LUAD. Moreover, elevated NRK predicted poor survival in LUAD patients.
Conclusion
Here, we first report that NRK is significantly elevated in LUAD-associated CAFs and may function as a promising therapeutic target for cancer combination treatment. Besides, modulation of ECM and glycolysis/gluconeogenesis pathways may be an efficient approach to alter CAFs functionality in LUAD.
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Data availability
All data generated or analyzed during this study are included in this published article and its supplementary information files.
Abbreviations
- CAFs:
-
Cancer-associated fibroblasts
- CCLE:
-
Cancer Cell Line Encyclopedia
- CSCs:
-
Cancer stem cells
- DEGs:
-
Differentially expressed genes
- ECM:
-
Extracellular matrix
- FC:
-
Fold change
- GO:
-
Gene Ontology
- IF:
-
Immunofluorescence
- KEGG:
-
Kyoto Encyclopedia of Genes and Genomes
- LUAD:
-
Lung adenocarcinoma
- LUSC:
-
Lung squamous cell carcinoma
- NFs:
-
Normal fibroblasts
- NRK:
-
Nik-related kinase
- NSCLC:
-
Non-small cell lung cancer
- RT-qPCR:
-
Real-time Quantitative Polymerase Chain Reaction
- SCLC:
-
Small cell lung cancer
- TME:
-
Tumor microenvironment
- WB:
-
Western Blot
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Acknowledgements
We express our gratitude to research team of Prof. Huaxin Hou and Prof. Danrong Li, who provided countless assistance during experiment process.
Funding
This study is funded by the National Natural Science Foundation of China (No. 81860649), Natural Science Foundation of Guangxi (No. 2018GXNSFAA050053), and Innovation Project of Guangxi Graduate Education (No. YCBZ2020054). This research was also funded by the first batch of cultivating talents of young and middle-aged backbone teachers in Guangxi universities and Guangxi First-class Discipline Project for Pharmaceutical Sciences (No. GXFCDP-PS-2018) and the Project of Innovation, Entrepreneurship, and Joint Training Base for Pharmaceutical Postgraduates.
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Experiments procedure and paper writing were conducted by TW. TW, JS and KL collected clinical specimens and pathological information, LL, XM, and NM helped to obtained signed informed consent from patients. Data were processed by TW, JS, and KL. ZH and GC offer excellent technical assistance. JY designed the study and revised the manuscript. All authors read and reached agreement on final manuscript.
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Patients signed the written informed consent and the study was approved by the Committee of Ethics, Guangxi Medical University, China.
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Wei, T., Song, J., Liang, K. et al. Identification of a novel therapeutic candidate, NRK, in primary cancer-associated fibroblasts of lung adenocarcinoma microenvironment. J Cancer Res Clin Oncol 147, 1049–1064 (2021). https://doi.org/10.1007/s00432-020-03489-z
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DOI: https://doi.org/10.1007/s00432-020-03489-z