CYP genes in osteosarcoma: Their role in tumorigenesis, pulmonary metastatic microenvironment and treatment response

// Alini Trujillo-Paolillo 1, 2 , Francine Tesser-Gamba 1 , Antonio Sergio Petrilli 3 , Maria Teresa de Seixas Alves 4 , Reynaldo Jesus Garcia Filho 5 , Renato de Oliveira 6 and Silvia Regina Caminada de Toledo 1, 2, 7 1 Genetics Laboratory, Pediatric Oncology Institute (IOP/GRAACC), Federal University of Sao Paulo, Rua Botucatu, Vila Clementino, Sao Paulo SP, 04023-062, Brazil 2 Department of Clinical and Experimental Oncology, Federal University of Sao Paulo, Rua Dr. Diogo de Faria, Vila Clementino, Sao Paulo SP, 04037-003, Brazil 3 Pediatric Oncology Institute (IOP/GRAACC), Department of Pediatrics, Federal University of Sao Paulo, Rua Botucatu, Vila Clementino, Sao Paulo SP, 04023-062, Brazil 4 Department of Pathology, Federal University of Sao Paulo, Rua Botucatu, Vila Clementino, Sao Paulo SP, 04023-062, Brazil 5 Department of Orthopedic Surgery and Traumatology, Federal University of Sao Paulo, Rua Borges Lagoa, Vila Clementino, Sao Paulo SP, 04038-031, Brazil 6 Department of Thoracic Surgery, Federal University of Sao Paulo, Rua Napoleao de Barros, Vila Clementino SP, 04024-002, Brazil 7 Department of Morphology and Genetics, Federal University of Sao Paulo, Rua Botucatu, Vila Clementino, Sao Paulo SP, 04023-062, Brazil Correspondence to: Silvia Regina Caminada de Toledo, email: silviatoledo@graacc.org.br Keywords: osteosarcoma, cytochrome P-450, osteosarcoma cell line, tumor microenvironment, treatment response Received: March 22, 2016     Accepted: January 06, 2017     Published: March 03, 2017 ABSTRACT Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. The present study investigated the expression of Cytochrome P-450 ( CYP ) genes: CYP1A2 , CYP3A4 and CYP3A5 by qRT-PCR in 135 specimens obtained from OS patients, including biopsy (pre-chemotherapy), tumor resected in surgery (post-chemotherapy), adjacent bone to tumor (nonmalignant tissue), pulmonary metastasis and adjacent lung to metastasis (nonmalignant tissue). Normal bone and normal lung tissues were used as control. We also investigated in five OS cell lines the modulation of CYPs expression by cisplatin, doxorubicin and methotrexate. As result, the adjacent lung specimens presented CYP1A2 overexpression compared to the normal lung (p=0.0256). Biopsy specimens presented lower CYP3A4 expression than normal bone (p=0.0314). The overexpression of both CYP1A2 and CYP3A4 in post-chemotherapy specimens were correlated with better event free-survival (p=0.0244) and good response (p=0.0484), respectively. Furthermore, in vitro assays revealed that CYP1A2 was upregulated by doxorubicin (p=0.0034); CYP3A4 was upregulated by cisplatin, doxorubicin and methotrexate (p=0.0004, p=0.0024, p<0.0001, respectively); and CYP3A5 was downregulated by doxorubicin (p=0.0285) and upregulated in time-dependent manner by methotrexate (p=0.0239). In conclusion, our findings suggest that CYP genes play an important role in OS tumorigenesis, at primary and metastatic sites, as well in treatment response.