TNBC cell lines (TNBCC), MDA-MB-231 and MDA-MB-468 were purchased (ATCC, CA, U.S.A.). The Gemcitabine-resistance cell lines (GRC), MDA-MB-231-R and MDA-MB-468-R were produced by treating TNBCC with increasing Gemcitabine concentrations (0.1–15 nm) for 12 months (GI50 < 7.0) [18 (link)]. Gemcitabine was bought from AbMole (Beijing, China). Cells were cultured in DMEM (PM150210) containing 4500 mg/l glucose, 10% fine FBS (164220-100) and 100 U/ml penicillin–streptomycin (PB180122) in 5% CO2, 95% O2 condition at 37°C (Procell, Wuhan, China).
Each cell line was also incubated with NLRP3 agonist Nigericin sodium salt (NSS) and antagonist CY-09 separately (Selleck, Shanghai, China) to differentiate NLRP3 expression in both TNBCC and GRC. Wnt inhibitor Wnt-C59 was added into subgroups of GRC so as to inactivate signaling pathway (Selleck, Shanghai, China).
All the cell lines were exposed to the 0, 1, 3, 5 nM Gemcitabine, respectively, for 72 h for the following assays.
Each cell line was also incubated with NLRP3 agonist Nigericin sodium salt (NSS) and antagonist CY-09 separately (Selleck, Shanghai, China) to differentiate NLRP3 expression in both TNBCC and GRC. Wnt inhibitor Wnt-C59 was added into subgroups of GRC so as to inactivate signaling pathway (Selleck, Shanghai, China).
All the cell lines were exposed to the 0, 1, 3, 5 nM Gemcitabine, respectively, for 72 h for the following assays.