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CMC is the most frequently diagnosed cancer and one of the leading causes of death in non-spayed female dogs. Exploring novel therapeutic agents is necessary to increase the survival rate of dogs with CMC. MPOBA is a BZOP derivative that has a significant anticancer effect in a human cell line. The main goal of this study was to investigate the anticancer properties of MPOBA against two CMC cell lines (REM134 and CMGT071020) using a 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, a wound healing assay, a transwell migration assay, an Annexin V-FITC apoptosis assay with a flow cytometry analysis, a mRNA expression analysis using quantitative real-time PCR (qRT-PCR), and an immunohistochemistry (IHC). According to the accumulated studies, MPOBA caused significant concentration- and time-dependent reductions in cell proliferation and cell migration and induced apoptosis in both CMC cell lines. In gene expression analysis, nine canine genes, including <i>TP53</i>, <i>BCL-2</i>, <i>BAX</i>, epidermal growth factor receptor (<i>EGFR</i>), snail transcription factor (<i>SNAIL</i>), snail-related zinc-finger transcription factor (<i>SLUG</i>), <i>TWIST</i>, <i>E-cadherin</i>, and <i>N-cadherin</i>, were investigated. The mRNA expression results revealed that MPOBA induced upregulation of <i>TP53</i> and overexpression of the pro-apoptotic gene <i>BAX</i>, together with an inhibition of <i>BCL-2.</i> Moreover, MPOBA also suppressed the mRNA expression levels of <i>SNAIL</i>, <i>EGFR</i>, and <i>N-cadherin</i> and induced upregulation of <i>E-cadherin</i>, crucial genes related to the epithelial-to-mesenchymal transition (EMT). However, there was no significant difference in the IHC results of the expression patterns of vimentin (VT) and cytokeratin (CK) between MPOBA-treated and control CMC cells. In conclusion, the results of the present study suggested that MPOBA exhibited significant anticancer activity by inducing apoptosis in both CMCs via upregulation of <i>TP53</i> and <i>BAX</i> and downregulation of <i>BCL-2</i> relative mRNA expression. MPOBA may prove to be a potential candidate drug to be further investigated as a therapeutic agent for CMC.