Abstract LB-A10: The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway

Abstract

Abstract The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway Yeo-Jung Kwon*, Sangyun Shin, Dong-Jin Ye, Hyoung-Seok Baek, and Young-Jin Chun Department of Pharmaceutical Biochemistry, College of Pharmacy, Chung-Ang University, Seoul, Korea Cytochrome P450 1B1 (CYP1B1) belongs to the CYP1 family and shares the feature as an enzyme for oxidative metabolisms. It has been reported that CYP1B1 expression is higher in the tumor tissues than the normal ones, especially in hormone-related cancers such as breast, ovarian, and prostate cancer cells. To explore the role of CYP1B1 on cancer cell proliferation, we studied the effects of CYP1B1 inhibition and activation in MCF-7 and MCF-10A cells. Here, we confirmed that expression of CYP1B1 affects the level of cell proliferation using CCK assay. To elucidate whether the cell proliferation induced by CYP1B1 is related to Wnt/β-catenin signaling, we examined the alteration of β-catenin and expression using western blot, RT-PCR, immunostaining, and qPCR followed by over-expression of CYP1B1 or treatment with DMBA (7,12-dimethylbenz[α]anthracene), a well known CYP1B1 inducer and these studies also performed when CYP1B1 gene was knockdown or cells were treated with TMS (tetramethoxystilbene), a specific CYP1B1 inhibitor. These investigations all showed that CYP1B1 promotes β-catenin expression in mRNA and protein levels and induces β-catenin nuclear localization. To identify the effect of CYP1B1 on Wnt/β-catenin signaling further, the promoter activity of β-catenin/TCF/LEF or expression level of Wnt/β-catenin signaling target genes such as c-myc, cyclin D1 were examined and all factors also showed to be up-regulated by CYP1B1. To elucidate whether CYP1B1 induces cell metastasis, cell migration and invasion assay were conducted and it has been identified that CYP1B1 enhances cell motility by induction of mesenchymal-like cell morphological changes. Moreover, the uPAR-uPA pathway has been identified to be related to the CYP1B1-induced cell metastasis by detection of uPAR-uPA pathway factors using western blot and qPCR in MCF-7 and HeLa cells. Taken together, these data suggest that CYP1B1 promotes cell proliferation and metastasis via inducing Wnt/β-catenin signaling and uPAR pathway. Citation Format: Yeo-Jung Kwon, Sangyun Shin, Dong-Jin Ye, Hyoung-Seok Baek, Young-Jin Chun. The carcinogenic role of CYP1B1 in cancer cells related to Wnt/β-catenin signaling and uPAR pathway. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr LB-A10.