MW‐19, a dihydropyrazole derivative, induces human triple‐negative breast cancer cell apoptosis by targeting apoptosis‐related pathways

Abstract

AbstractPrevious studies have indicated that heterocyclic substituted dihydropyrazole derivatives, particularly MW‐19, potentially exert anticancer activity in vitro; however, the underlying mechanism remains unknown. The present study was designed to investigate the mechanisms underlying MW‐19 activity in triple‐negative breast cancer cells. A sulforhodamine B assay was performed to evaluate cell proliferation inhibition rates, and the antitumor effect of MW‐19 was evaluated in mice with HCC‐1806 xenografts. Apoptosis was analyzed by Hoechst 33342 and annexin V/propidium iodide staining. Expression of pro‐ and antiapoptotic proteins and mRNA were analyzed by western blotting and reverse transcription‐quantitative (RT‐q) PCR, respectively. We found that MW‐19 significantly inhibited HCC‐1806 cell proliferation in a dose‐ and time‐dependent manner, and significantly inhibited MDA‐MB‐231 cell migration. Importantly, oral administration of MW‐19 significantly inhibited HCC‐1806 tumor growth in BALB/c‐nu/nu mice. Moreover, MW‐19 treatment induced marked apoptosis and G2/M arrest in the sensitive cell line, HCC‐1806. RT‐qPCR analysis showed that levels of proapoptotic genes (Bax, caspase‐3, caspase‐7, and Fas) were considerably increased in the MW‐19 group relative to the control group, while those of antiapoptotic factors (Bcl‐2, C‐MYC) were dramatically decreased. Consistently, Bax, caspase‐3, and caspase‐7 were significantly induced after MW‐19 treatment, while levels of phosphorylated (p‐)AKT, p‐PI3K, p‐ERK, and the antiapoptotic protein, Bcl‐2, were clearly diminished, and the P38 MAPK signaling pathway was activated. Furthermore, P38 pharmacological inhibitors abrogated MW‐19‐induced apoptosis. Together, our findings indicate that MW‐19 exerts antitumor effects by targeting PI3K/AKT and ERK/P38 signaling pathways.