Hcc827

Human lung adenocarcinoma A549 (ATCC Cat# CRL-7909), HCC827 (KCLB Cat# 70827), H2444 (ATCC Cat# CRL-5945), H2030 (ATCC Cat# CRL-5914), and H1975 (ATCC Cat# CRL-5908) cell lines were obtained from Cobioer Biosciences (Nanjing, China). Short tandem repeat (STR) analyses were performed as follows: the analyses of the A549, HCC827, and H1975 cell lines were performed in 2017, and the analyses of the H2444 and H2030 cell lines were performed in 2019. A549, H2444, and H2030 cells harbor mutations in KRAS, and H1975 and HCC827 cells harbor EGFR mutations. The A549 cells were cultured in F12K medium containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA), 100 U/ml penicillin and 0.1 mg/ml streptomycin (HyClone, Logan, NT, USA). The other cell lines were cultured in RPMI 1640 (HyClone) containing 10% fetal bovine serum supplemented with 100 U/ml penicillin and 0.1 mg/ml streptomycin. The cells were maintained at 37°C in a humidified incubator with 5% CO₂. All cell lines were confirmed to be Mycoplasma-negative (Biothrive Sci. & Tech. Ltd., Shanghai, China), and the cell lines were used within 3 months after resuscitation.

Human lung adenocarcinoma A549, H1975, and HCC827 cells were obtained from Cobioer Biosciences (Nanjing, China). Authentication of these cell lines was performed by short tandem repeat (STR) DNA profiling by Cobioer Biosciences in July 2017. All of the cell lines were passaged for fewer than four months. The cells were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (GIBCO, Grand Island, NY, USA) and 1% penicillin/streptomycin (Hyclone, Logan UT, USA). Lung cancer cells were treated with PGE₂ (MedChem Expression, Monmouth Junction, NJ, USA) or Sulprostone, Butaprost and CAY10598 (Cayman Chemical, Ann Arbor, MI, USA), which are selective agonists for EP1/EP3, EP2, and EP4, respectively. The inhibitors celecoxib (COX-2), GDC-0068 (AKT) and LY294002 (PI3K) were obtained from MedChem Express. All of the EP agonists and inhibitors were dissolved in dimethyl sulfoxide (DMSO). The final concentration of DMSO in cell culture did not exceed 0.1% (v/v). The cells in the experiments that were treated with DMSO alone served as the vehicle control (blank).