Evaluation of the effects of Cinobufagin on G361 Melanoma Cell Cultures and a comparison with paclitaxel.

Abstract

Abstract The melanoma pathophysiology is considerably complex. Wnt signaling via the β-catenin/transcription factor 7-like 2 (TCF4)/Lymphoid enhancer-binding factor 1 (LEF1) complex, the microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-2 (TRP-2), tyrosinase (Tyr) and cyclin-dependent kinase 2 (Cdk2) are reported to activate the transcriptional gene expression associated with pigmentation and the differentiation and proliferation of melanocytes and malignant melanoma cells. Cinobufagin is a leading active ingredient in Traditional Chinese Medicine, and has been approved in China as a chemotherapeutic agent for the treatment of liver and prostate cancer. The primary aim of the present study is to evaluate the effects of cinobufagin and paclitaxel on melanoma cells, both individually and in combination, in the G361 melanoma cell line. Within the scope of the study, IC50 doses were determined based on an MTT analysis, Caspase-3 measurements were analyzed using the ELISA method and mRNA expression levels were analyzed using the RT PCR method. Cinobufagin used in combination with paclitaxel was found to increase Caspase-3 levels more than when cinobufagin was used alone, and it was further determined that cinobufagin treatment decreased the expression levels of the β-catenin, C-myc and Cyclin D1 genes, while paclitaxel and cinobufagin used in combination were found to increase all mRNA expression levels (Bax, Caspase-3, Bcl2, β-catenin, C-myc, Wnt and Cyclin D1). Cinobufagin can be considered a promising natural pharmaceutical agent for the targeted treatment of cancers with high levels of LEF1.