Comprehensive Analysis of Breast Cancer Cell Lines

The partitions (AC n-hexane, IC n-hexane, and IC ethylacetate), and the AC n-hexane-ethylacetate fraction were obtained from our previous study [8 (link)]. The HPLC grade of methanol, acetonitrile, and phosphate buffer pH 7.4 were used as the mobile phase, whereas the stationary phase used C₁₈ column. The MMP9 enzyme kit was obtained from BioVision and comprised lyophilized MMP9, FRET-based MMP9 substrate (Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg), MMP9 assay buffer, and NNGH inhibitor (N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycyl hydroxamic acid) as its positive control. MDA-MB-231, MCF7, T47D, and 4T1 cells were obtained from Parasitology Laboratory, Medical Faculty, Gadjah Mada University), and cultured in Dulbecco’s Modified Eagle Media (DMEM) for MDA-MB-231, MCF7, and T47D, and RPMI-1640 medium containing 10% (v/v) fetal bovine serum (FBS) and 1% penicillin–streptomycin was used as the media for 4T1 (Life Technologies, Carlsbad, CA, USA) at 37 °C in a 5% CO₂ humidified incubator. RNase was courtesy from Parasitology Laboratory, Medical Faculty, Gadjah Mada University and cultured in Dulbecco’s Modified Eagle Media (DMEM). Doxorubicin (DOX) and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl tetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA).