Hek 293

Human embryonic kidney (HEK)-293 and Michigan Cancer Foundation (MCF) -7 cell lines were purchased from National Centre for Cell Science (Pune, India). Dulbecco modified Eagle’s media (DMEM) was obtained from Himedia (Mumbai, India), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue from Sigma Aldrich (Mo, USA), fetal bovine serum (FBS), streptomycin-penicillin (Strep-Pen), trypsin/EDTA solution and dimethyl sulfoxide (DMSO) were procured from Thermo Fischer (MA, USA).

Fetal bovine serum was obtained from Sigma Aldrich. All the solvents, chemicals, and reagents were purchased from Hi Media laboratories (Mumbai). Cancer cell lines viz., MCF 7 (human breast cancer), HEP G2 (liver cancer), NCIH 460 (non-small cell lung cancer), and non-cancerous human kidney cell line, HEK293 were procured from the National Centre for Cell Sciences (NCCS), Pune.

The Luria broth and Luria agar were taken from BD Difco™ Franklin Lakes, NJ. Human cell lines (MCF-7, A549 and HEK-293) used were procured from National Centre for Cell Sciences, Pune, India. Phospho-Tau, MARK4 (MA5-27002). actin monoclonal antibodies, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-iphenyltetrazolium bromide), antibiotic cocktail, fetal bovine serum, Dulbecco's modified eagle's media (DMEM) and propidium iodide (PI) were procured from Gibco-life technologies, Thermo Fischer Scientific (USA). All other reagents were of analytical grade purchased from local supplier.

Human KB 3–1, KB Ch^R 8–5, SW480 and HEK 293 cells were obtained from the National Centre for Cell Science (NCCS), Pune, India. The cells were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 mg/ml), 1% L-glutamine and 1% non-essential amino acids at 37 °C in a humidified atmosphere and 5% CO₂. The resistant KB Ch^R 8–5 cells were derived from the parental KB cells and were continuously grown in culture media containing 10 nM of colchicine in order to maintain the resistance. The resistant SW480-VCR cells were obtained by continuous exposure of parental SW480 cells to vincristine from a starting concentration of 0.625 nM. When the cells were able to grow at a given concentration of vincristine, they were harvested and plated in a new flask and exposed to a 2-fold higher concentration of vincristine. Likewise, the cells were exposed to higher concentration of vincristine and after 3 months of continuous exposure, the final obtained resistant SW480-VCR cells could able to grow in the presence of 20 nM of vincristine. The cells were cultured in tissue culture flasks. When they reached confluence, they were harvested and plated in new flasks at 1:2 ratios. Up to 10–15 passages were used for drug treatment.