Lncap

Two PCa cell lines viz., androgen-dependent LNCaP cell line, and androgen-independent PC-3 cell line were selected for the study, which broadly represents the clinical scenario of PCa stages. The LNCaP cell line is androgen-sensitive, less malignant, less metastatic, and possess epithelial features. PC-3 cells are androgen independent, malignant, metastatic, and possess mesenchymal features when compared to other PCa cell lines such as LNCaP, VCaP, and RWPE1. The PC-3 and LNCaP human prostate adenocarcinoma cell lines were obtained from National Centre for Cell Science (NCCS), Pune, and maintained in RPMI-1640 supplemented with 10% fetal bovine serum, penicillin, and streptomycin (Himedia Laboratories Pvt. Ltd., India). Both the cell lines were then subjected to doses of human recombinant TGF-β (Himedia Laboratories Pvt. Ltd., India) for inducing EMT. The treatment dosages of TGF-β were calculated post-MTT assay for IC50 evaluations. All the experiments were performed in triplicates.

The prostate carcinoma cell lines (AR-positive LNCaP and AR-negative PC-3) and noncancer cell lines (CHO and COS-1) were obtained from the National Centre for Cell Science (NCCS), Pune, India. PC-3, CHO and COS-1 cells were maintained in Dulbecco’s modified Eagle’s media (DMEM) while LNCaP cells were maintained in RPMI-1640 medium supplemented with 10% foetal bovine serum (FBS) (heat inactivated) under 5% CO₂ at 37°C. All the experiments were performed using LNCaP, PC-3, CHO and COS-1 cells from passage below 29, 34, 20 and 30 respectively. The cells were washed properly before changing the media to steroid-free complete media prior to each treatment with the compounds, unless otherwise stated.