Cell proliferation was evaluated by MTT assay, MMPs by gelatinase zymography, invasion through Matrigel, morphology by H&E, and Pgp expression by Western blot and immunodetection using FITC-conjugated antibody and rhodamine-123 accumulation and efflux assays. NM exhibited antiproliferative effect on MES-SA/Dx5, by 20% at 50 and 100 μg/ml and by 36%, 40% and 48% at 250, 500 and 1000 μg/ml, respectively. In contrast, NM treatment of MES-SA cells resulted in significantly increased cytotoxicity: 40%, 46%, 65% and 72% at 50, 100, 500 and 1000 μg/ml, respectively. In both cell lines, zymography demonstrated a band corresponding to MMP-2 in normal cells and MMP-9 with PMA treatment. Both MMPs showed dose-response inhibition by NM. NM treatment also showed diminished dose-dependent Pgp expression by MES-SA/Dx5 cell line by Western blot and by immunodetection, whereas MES-SA did not exhibit Pgp by Western blot or by immunostaining. NM enhanced the accumulation and efflux of Pgp substrate Rh-123 in MES-SA/Dx5 uterine sarcoma cell line but not in the drug-sensitive cell line MES-SA. In conclusion, NM demonstrated potent anticancer effects in both the drug-resistant and sensitive cell lines and modulated Pgp, suggesting its potential therapeutic effect in drug resistant, as well as sensitive cancers.
nutrients; multidrug resistant uterine sarcoma; MES-SA; MES-SA/Dx5; p-glycoprotein; MMPs; cytotoxicity; Matrigel invasion