Abstract
Purpose
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.
Methods
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.
Results
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.
Conclusions
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.
Similar content being viewed by others
Abbreviations
- RCC:
-
Renal cell carcinoma
- CB2:
-
Cannabinoid receptor 2
- ccRCC:
-
Clear-cell renal cell carcinoma
- OS:
-
Overall survival
- TMA:
-
Tissue microarrays
- FBS:
-
Fetal bovine serum
- DMSO:
-
Dimethyl sulfoxide
- SRB:
-
Sulforhodamine B
- C-index:
-
Harrell’s concordance index
- AIC:
-
Akaike information criteria
- IHC:
-
Immunohistochemistry
References
Brown I, Cascio MG, Rotondo D, Pertwee RG, Heys SD, Wahle KW (2013) Cannabinoids and omega-3/6 endocannabinoids as cell death and anticancer modulators. Prog Lipid Res 52(1):80–109
Caffarel MM, Andradas C, Mira E, Perez-Gomez E, Cerutti C, Moreno-Bueno G et al (2010) Cannabinoids reduce ErbB2-driven breast cancer progression through Akt inhibition. Mol Cancer 9:196
Chan PC, Sills RC, Braun AG, Haseman JK, Bucher JR (1996) Toxicity and carcinogenicity of delta 9-tetrahydrocannabinol in Fischer rats and B6C3F1 mice. Fundam Appl Toxicol 30(1):109–117
Chow WH, Dong LM, Devesa SS (2010) Epidemiology and risk factors for kidney cancer. Nat Rev Urol 7(5):245–257
De Jesus ML, Hostalot C, Garibi JM, Salles J, Meana JJ, Callado LF (2010) Opposite changes in cannabinoid CB1 and CB2 receptor expression in human gliomas. Neurochem Int 56(6–7):829–833
Elbaz M, Ahirwar D, Ravi J, Nasser MW, Ganju RK (2016) Novel role of cannabinoid receptor 2 in inhibiting EGF/EGFR and IGF-I/IGF-IR pathways in breast cancer. Oncotarget 8(18):29668–29678
Elmore JM, Kadesky KT, Koeneman KS, Sagalowsky AI (2003) Reassessment of the 1997 TNM classification system for renal cell carcinoma. Cancer 98(11):2329–2334
Engeli S (2012) Central and peripheral cannabinoid receptors as therapeutic targets in the control of food intake and body weight. Handb Exp Pharmacol 209:357–381
Felder CC, Glass M (1998) Cannabinoid receptors and their endogenous agonists. Annu Rev Pharmacol Toxicol 38:179–200
Ficarra V, Novara G, Galfano A, Brunelli M, Cavalleri S, Martignoni G et al (2009) The ‘Stage, Size, Grade and Necrosis’ score is more accurate than the University of California Los Angeles Integrated Staging System for predicting cancer-specific survival in patients with clear cell renal cell carcinoma. BJU Int 103(2):165–170
Gupta K, Miller JD, Li JZ, Russell MW, Charbonneau C (2008) Epidemiologic and socioeconomic burden of metastatic renal cell carcinoma (mRCC): a literature review. Cancer Treat Rev 34(3):193–205
Guzman M, Sanchez C, Galve-Roperh I (2002) Cannabinoids and cell fate. Pharmacol Ther 95(2):175–184
Hang J, Hu H, Huang J, Han T, Zhuo M, Zhou Y et al (2016) Sp1 and COX2 expression is positively correlated with a poor prognosis in pancreatic ductal adenocarcinoma. Oncotarget 7(19):28207–28217
Hong SK, Jeong CW, Park JH, Kim HS, Kwak C, Choe G et al (2011) Application of simplified Fuhrman grading system in clear-cell renal cell carcinoma. BJU Int 107(3):409–415
Hu G, Ren G, Shi Y (2011) The putative cannabinoid receptor GPR55 promotes cancer cell proliferation. Oncogene 30(2):139–141
Jia N, Zhang S, Shao P, Bagia C, Janjic JM, Ding Y et al (2014) Cannabinoid CB2 receptor as a new phototherapy target for the inhibition of tumor growth. Mol Pharm 11(6):1919–1929
Klein Nulent TJ, Van Diest PJ, van der Groep P, Leusink FK, Kruitwagen CL, Koole R et al (2013) Cannabinoid receptor-2 immunoreactivity is associated with survival in squamous cell carcinoma of the head and neck. Br J Oral Maxillofac Surg 51(7):604–609
Larrinaga G, Sanz B, Perez I, Blanco L, Candenas ML, Pinto FM et al (2010) Cannabinoid CB(1) receptor is downregulated in clear cell renal cell carcinoma. J Histochem Cytochem 58(12):1129–1134
Larrinaga G, Sanz B, Blanco L, Perez I, Candenas ML, Pinto FM et al (2013) Cannabinoid CB1 receptor is expressed in chromophobe renal cell carcinoma and renal oncocytoma. Clin Biochem 46(7–8):638–641
Ligresti A, Bisogno T, Matias I, De Petrocellis L, Cascio MG, Cosenza V et al (2003) Possible endocannabinoid control of colorectal cancer growth. Gastroenterology 125(3):677–687
Ligresti A, De Petrocellis L, Di Marzo V (2016) From phytocannabinoids to cannabinoid receptors and endocannabinoids: pleiotropic physiological and pathological roles through complex pharmacology. Physiol Rev 96(4):1593–1659
Liu CS, Chau SA, Ruthirakuhan M, Lanctot KL, Herrmann N (2015) Cannabinoids for the treatment of agitation and aggression in Alzheimer’s Disease. CNS Drugs 29(8):615–623
Ljungberg B, Campbell SC, Choi HY, Jacqmin D, Lee JE, Weikert S et al (2011) The epidemiology of renal cell carcinoma. Eur Urol 60(4):615–621
Ljungberg B, Bensalah K, Canfield S, Dabestani S, Hofmann F, Hora M et al (2015) EAU guidelines on renal cell carcinoma: 2014 update. Eur Urol 67(5):913–924
Malik Z, Baik D, Schey R (2015) The role of cannabinoids in regulation of nausea and vomiting, and visceral pain. Curr Gastroenterol Rep 17(2):429
Martínez-Martínez E, Gómez I, Martín P, Sánchez A, Román L, Tejerina E et al (2015) Cannabinoids receptor type 2, CB2, expression correlates with human colon cancer progression and predicts patient survival. Oncoscience 2(2):131–141
Martínez-Martínez E, Martín-Ruiz A, Martín P, Calvo V, Provencio M, García JM (2016) CB2 cannabinoid receptor activation promotes colon cancer progression via AKT/GSK3β signaling pathway. Oncotarget 7(42):68781–68791
McKallip RJ, Nagarkatti M, Nagarkatti PS (2005) Delta-9-tetrahydrocannabinol enhances breast cancer growth and metastasis by suppression of the antitumor immune response. J Immunol 174(6):3281–3289
Mills B, Yepes A, Nugent K (2015) Synthetic cannabinoids. Am J Med Sci 350(1):59–62
Palazzo E, Luongo L, Novellis V, Rossi F, Maione S (2010) The role of cannabinoid receptors in the descending modulation of pain. Pharmaceuticals (Basel) 3(8):2661–2673
Patard JJ, Kim HL, Lam JS, Dorey FJ, Pantuck AJ, Zisman A et al (2004) Use of the University of California Los Angeles integrated staging system to predict survival in renal cell carcinoma: an international multicenter study. J Clin Oncol 22(16):3316–3322
Perez-Gomez E, Andradas C, Blasco-Benito S, Caffarel MM, Garcia-Taboada E, Villa-Morales M et al (2015) Role of cannabinoid receptor CB2 in HER2 pro-oncogenic signaling in breast cancer. J Natl Cancer Inst 107(6):077
Reimers M, Carey VJ (2006) Bioconductor: an open source framework for bioinformatics and computational biology. Methods Enzymol 411:119–134
Ross RA, Brockie HC, Stevenson LA, Murphy VL, Templeton F, Makriyannis A et al (1999) Agonist-inverse agonist characterization at CB1 and CB2 cannabinoid receptors of L759633, L759656, and AM630. Br J Pharmacol 126(3):665–672
Sarfaraz S, Afaq F, Adhami VM, Mukhtar H (2005) Cannabinoid receptor as a novel target for the treatment of prostate cancer. Cancer Res 65(5):1635–1641
Siegel RL, Miller KD, Jemal A (2016) Cancer statistics, 2016. CA Cancer J Clin 66(1):7–30
Smaletz O (2015) Current management and future directions in the treatment of advanced renal cell carcinoma-a latin american perspective: 10 years in review. Int Braz J Urol 41(5):835–843
Velasco G, Hernandez-Tiedra S, Davila D, Lorente M (2016) The use of cannabinoids as anticancer agents. Prog Neuropsychopharmacol Biol Psychiatry 64:259–266
Vemuri VK, Makriyannis A (2015) Medicinal chemistry of cannabinoids. Clin Pharmacol Ther 97(6):553–558
Waalkes S, Kramer M, Herrmann TR, Schrader AJ, Kuczyk MA, Merseburger AS (2010) Present state of target therapy for disseminated renal cell carcinoma. Immunotherapy 2(3):393–398
Wang L, Ma L, Wang X, Li B, Guo S, Qiao Q (2015) Therapeutic effects and associated adverse events of first-line treatments of advanced renal cell carcinoma (RCC): a meta-analysis. Int Urol Nephrol 47(4):617–624
Xu X, Liu Y, Huang S, Liu G, Xie C, Zhou J et al (2006) Overexpression of cannabinoid receptors CB1 and CB2 correlates with improved prognosis of patients with hepatocellular carcinoma. Cancer Genet Cytogenet 171(1):31–38
Zurier RB, Burstein SH (2016) Cannabinoids, inflammation, and fibrosis. Faseb J 30(11):3682–3689
Acknowledgements
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.
Author information
Authors and Affiliations
Contributions
Jianfeng Wang, Yunze Xu, and Liangsong Zhu conceived and designed the experiments. Jianfeng Wang the manuscript and performed the experiments. Jianfeng Wang and Yunze Xu performed the immunohistochemistry assay. Jianfeng Wang and Liangsong Zhu performed the RNA interference (RNAi) transfection. Jianfeng Wang and Yun Zou performed the proliferative ability analysis. Jianfeng Wang, Wen Kong, Baijun Dong, and Jiwei Huang performed cell migration assay. Jianfeng Wang, Yonghui Chen and Wei Xue performed cell cycle analysis and statistical analysis. Yiran Huang and Jin Zhang oversight of all aspects of the study. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Ethics statement
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.
Electronic supplementary material
Below is the link to the electronic supplementary material.
432_2017_2527_MOESM1_ESM.tif
S1 Figure. Expression of CB2 is elevated in RCC tissues. 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)
432_2017_2527_MOESM2_ESM.tif
S2 Figure. Inhibition of CB2 suppresses the migration and invasion ability of RCC cells. (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)
432_2017_2527_MOESM3_ESM.tif
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)
432_2017_2527_MOESM4_ESM.tif
S4 Figure. Inhibition of CB2 has little effect on the proliferation of HK-2. 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)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-017-2527-y