Abstract
Background
Retinoic acid-related orphan receptor C isoform 2 (RORC2) is regarded as a pathogenic factor for autoimmune and inflammatory diseases and tumours. Previous studies have primarily focused on RORC2 expression in IL-17-producing immune cells but not in carcinoma cells; thus, little is known about the roles of RORC2 in the progression of human non-small cell lung cancer (NSCLC). In this study, we analysed the expression of RORC2 and its participation in tumour progression in NSCLC.
Methods
RORC2 expression in NSCLC and adjacent normal lung tissues was assessed via quantitative real-time PCR (qRT-PCR) and immunohistochemistry. RORC2 expression in NSCLC cell lines was examined by qRT-PCR, Western blotting and flow cytometry. The effects of inhibiting RORC2 activity on the proliferation of NSCLC cells were evaluated. The prognostic value of RORC2 for NSCLC was revealed based on Kaplan–Meier analysis.
Results
High RORC2 expression was observed in lung cancer tissues and was significantly related to age (p = 0.013) and regional lymph node metastasis (p = 0.009). RORC2 expression was higher in the A549, H460, SPC-A1 and H1299 cell lines than in a control cell line. In addition, cell proliferation was decreased in NSCLC cells upon the blocking of RORC2 activity using a specific inhibitor. High RORC2 expression correlated with worse overall survival (p = 0.030).
Conclusions
Our study suggests that RORC2 is expressed by lung cancer cells and greatly contributes to tumour cell proliferation and overall survival in NSCLC. These findings strongly imply that RORC2 is associated with tumour progression.
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Acknowledgments
The research was supported by the National Natural Science Foundation of China (No. 81072400), the Research Fund for the Doctoral Program of Higher Education of China (No. 20130142110066) and the Natural Science Foundation of Hubei Province (No. 2014CFA057).
Author contributions
Yang Jin was the designer and the leader of the project; Qi Huang and Jinshuo Fan participated in drafting the manuscript and carried out the immunoassays; Xin Qian, Jiao Du and Caiyun Chen participated in the design and performed the statistical analysis; Xiuxiu Zhang, Zhilei Lv and Jieli Han participated in qRT-PCR. Mengfei Guo, Feng Wu and Guorong Hu participated in Western blotting and flow cytometry. All authors have read and approved the final manuscript.
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the Union Hospital, Tongji Medical College, and Huazhong University of Science and Technology ([2013]IEC(S202)) ethical committees and with the 1964 Helsinki Declaration and its later amendments.
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Informed consent was obtained from all individual participants included in the study.
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Qi Huang, Jinshuo Fan and Xin Qian have contributed equally to this work.
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432_2015_2040_MOESM1_ESM.tif
Supplementary Figure 1 Positive staining for RORC2 was primarily detected on the surface of carcinoma cells and occasionally in non-tumour cells such as infiltrating immune cells in tumour tissues (magnification, 200×). (TIFF 20952 kb)
432_2015_2040_MOESM2_ESM.tif
Supplementary Figure 2 RORC2 inhibition results in decreased IL-17 expression. (A) and (B) Decreased IL-17 protein production was observed in A549 and H292 cells based on Western blotting. (C) and (D) Grey density analysis using the ImageJ software for the results shown in (A) and (B), respectively. (TIFF 31096 kb)
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Huang, Q., Fan, J., Qian, X. et al. Retinoic acid-related orphan receptor C isoform 2 expression and its prognostic significance for non-small cell lung cancer. J Cancer Res Clin Oncol 142, 263–272 (2016). https://doi.org/10.1007/s00432-015-2040-0
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DOI: https://doi.org/10.1007/s00432-015-2040-0