Renal cell carcinoma (RCC) is the most common malignancy of urogenital system, and patients with RCC may face a poor prognosis. However, limited curable therapeutic options are currently available. The aim of this study is to investigate the role of Cannabinoid receptor 2 (CB2) in RCC progression.
Immunohistochemistry was to investigate the expression pattern of CB2 in 418 RCC tissues and explore its prognostic function in RCC patients. Furthermore, the role of used CB2 si-RNA knockdown and inhibited by AM630, a CB2 inverse agonist, on cell proliferation, migration, and cell cycle of RCC cell lines in vitro was also investigated.
We observed that CB2 was up-regulated in RCC tissues, and presented as an independent prognostic factor for overall survival of RCC patients and higher CB2 expression tends to have poor clinical outcomes in survival analyses. Moreover, we also observed that CB2, incorporated with pN stage, pathological grade, and recurrence or distant metastasis after surgery, could obviously enhance their prognostic accuracy in a predictive nomogram analysis. In addition, knockdown or inhibition by AM630 for the expression of CB2 in vitro could significantly decreased cell proliferation and migration, and obviously induced cell cycle arrest in G2/M of RCC cells.
CB2 expression is functionally related to cellular proliferation, migration, and cell cycle of RCC cells. Our data suggest that CB2 might be a potential therapeutic target for RCC.
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Renal cell carcinoma
Cannabinoid receptor 2
Clear-cell renal cell carcinoma
Fetal bovine serum
Harrell’s concordance index
Akaike information criteria
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This work was supported by National Natural Science Foundation of China (nos. 81472378, 81272841, and 91129725), Shanghai Committee of Science and Technology (13ZR1425100). All these study sponsors have no roles in the study design, in the collection, analysis, and interpretation of data.
Conflict of interest
The authors declare that they have no competing interests.
This investigation was approved by the Ethics and Research Committees of Renji Hospital, Shanghai Jiao Tong University School of Medicine, and was conducted in accordance with the ethical standards and according to the Declaration of Helsinki and according to national and international guidelines. Tissue samples were obtained with written consent from all the patients.
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CB2 mRNA expression analyzed by real-time RT-PCR in RCC tissues. CB2 mRNA expression levels were normalized to GAPDH mRNA expression. CB2 mRNA was determined to be higher in the RCC tissues compared to adjacent normal tissues. (TIFF 39 kb)
(A and B) AM630 reduced the number of invasion cells in 786-O cell line at 10 μM and 20 μM concentrations (Bar: 1 mm); (C and D) 786-O cells were seeded on six-well plates. A single scratch was made after the cells grew about 90% confluence. After treatment of AM630 for 6 h, the cells were photographed (20 ×). The lines indicate that the area occupied by the initial scraping was quantified. (TIFF 21727 kb)
S3 Figure. Knockdown of CB2 significantly reduced the levels of cyclin B1 and cdc-25c expression in RCC cells.
Western blotting analysis revealed that knockdown of CB2 significantly reduced the expression levels of cyclin B1 and cdc-25c in 786-O and CAKI-1 cells. (TIFF 1643 kb)
HK-2 cells were treated at the indicated concentrations, and the cell viability was analyzed by SRB assay. AM630 shows little effect on cell proliferation ability of HK-2. (TIFF 20 kb)
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Wang, J., Xu, Y., Zhu, L. et al. Cannabinoid receptor 2 as a novel target for promotion of renal cell carcinoma prognosis and progression. J Cancer Res Clin Oncol 144, 39–52 (2018). https://doi.org/10.1007/s00432-017-2527-y
- Renal cell carcinoma
- Cannabinoid receptor 2