Docetaxel

Docetaxel and Nanoxel were purchased from Tokyo Chemical Industry (Tokyo, Japan) and Samyang Biopharmaceuticals (Seongnam, Korea), respectively. Thrombin, 5-hydroxytryptamine (5-HT), menadione, and butylated hydroxyanisole (BHA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Collagen was acquired from Chrono-log (Havertown, PA, USA) and 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST-1) was obtained from Seoul Clinical Genomics (Seoul, Korea). Fura-2/AM and 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red) were purchased from Molecular Probes (Eugene, OR, USA). Premix WST-1 Cell Proliferation Assay System was acquired from Takara Bio (Shiga, Japan). All other chemicals used were of the highest purity available and purchased from standard suppliers.

A2780 and SK‐OV‐3 cell lines were purchased from the ATCC and maintained in RPMI‐1640 (Nacalai Tesque) containing 10% FBS and antibiotics. The mirVana miRNA mimics for miR‐187‐5p (ID: MC12652), miR‐1908‐5p (ID: MC13846), miR‐6870‐5p (ID: MC27099), and negative control #1 were purchased from Thermo Fisher Scientific. Cells were seeded in 96‐well plates and transfected with 20 nM mimic using Lipofectamine RNAi Max (Thermo Fisher Scientific). After 24, 48, and 72 h of incubation, cell viability was assessed using CellTiter‐Glo 2.0 Cell Viability Assay (Promega) and a microplate reader (Gen5 Synergy H4; BioTek). For drug sensitivity analysis, after 24 h of transfection, the culture medium was replaced with cisplatin (Nichi‐Iko Pharmaceutical) or docetaxel (Tokyo Chemical Industry) containing medium, and cells were incubated for 48 h. Cell viability was then assessed using CellTiter‐Glo 2.0 Cell Viability Assay.
To evaluate transfection efficacy, quantitative RT‐PCR was carried out. Total RNA was extracted using the miRNeasy Mini Kit (Qiagen), and cDNA was synthesized using the TaqMan Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific). TaqMan Fast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan Advanced miRNA Assays (assay IDs 479423_mir, 478735_mir, and 480864_mir; Thermo Fisher Scientific) were used for quantitative RT‐PCR.