Cisplatin

Hepatic cancer cell lines HepG2, Huh-7, and Hepa1-6 were obtained from the Chinese Academy of Sciences (Shanghai, China). All cells were cultured at 37 °C in a humidified incubator containing 5% CO₂ using Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, New York, NY, USA) supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (Invitrogen, Waltham, MA, USA) and 10% fetal bovine serum (FBS) (Sigma, Australia). The trypsin (Gibco, USA) was used during cell passage. In some experiments, the cells were treated with the CHK1 inhibitor prexasertib (LY2603618) purchased from MedChemExpress (Monmouth Junction, NJ, USA) or chemotherapeutic reagent cisplatin from APExBIO (Houston, TX, USA).

Nicotinamide was purchased from Sigma-Aldrich (Sigma 72345). Cisplatin was purchased from APExBIO(A8321).

Compounds used included trametinib (Selleck Chemicals, Houston, TX, USA), Z-VAD-FMK (UBP Bio, Aurora, CO, USA), Nec-1 (ApexBio, Houston, TX, USA), cisplatin (ApexBio), and staurosporine (ApexBio). trametinib, Z-VAD-FMK, Nec-1, and staurosporine were dissolved in DMSO (Fisher Scientific, Waltham, MA, USA) and cisplatin was dissolved in sterile water. Control samples in all experiments performed were treated with vehicle only. Vehicle concentration in growth medium did not exceed 0.2%. Cell treatment was performed by aspirating the growth media from the cells and replacing it with growth medium containing selected concentrations of drugs.

Male six‐week‐old BALB/c nude mice weighing around 18.30 g were acquired from the Experimental Animal Center of Anhui Medical University (Hefei). The nude mice were kept at 20‑26°C, 40‑70% humidity, a 12/12 light/dark cycle, in a pathogen‑free environment and with free access to water and food. For the subcutaneous transplantation model, the mice were implanted with sh‑TAK1‑luciferase or sh‑NC‑luciferase GC cells (2.0 × 10⁶) in the right groin. The mice were killed 4 weeks after implantation, and the tumour volume was calculated. For the tail vein xenograft model, the mice in every group were administered with sh‑TAK1‑luciferase or sh‑NC‑luciferase GC cells (2.0 × 10⁶ suspended in 200 μL PBS) by the tail vein and killed 5 weeks later. Lung nodules and progression were monitored and quantified by bioluminescence. For tumour drug resistance evaluation, each group received an intraperitoneal injection of 5‐FU (cat. no. A4071, 40 mg/kg bodyweight, ApexBio) or cisplatin (cat. no. A8321, 4 mg/kg bodyweight, ApexBio) twice a week for 3 weeks. All procedures with animals have been endorsed by the Ethics Committee of The Fourth Affiliated Hospital of Anhui Medical University (certification no. LLSC20150234).

Human EGFR-TKIs resistant NSCLC cell line NCI-H1975 (#No. CRL-5908TM) was purchased from ATCC (American type culture collection; Manassas, VA, USA). Immortalized human epithelial cell line BEAS-2B was also obtained from ATCC. Human lung squamous cell carcinoma and immortalized human liver cell line 95-D and HL7702, respectively, were purchased from Shanghai cell bank affiliated to the Chinese Academy of Sciences (Shanghai, China). H1975 and HL7702 cells were maintained in RPMI-1640 medium (Sigma, St. Louis, MO, USA) containing 10% fetal bovine serum (FBS, Gibco, USA). BEAS-2B and 95-D cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Sigma, St. Louis, MO, USA) supplemented with 5 and 10% fetal bovine serum, respectively, in a humidified atmosphere containing 5% CO₂ at 37°C.
Osimertinib (AZD-9291), 10058-F4 (Myc-Max disruptor), and STF-31 (a specific Glut-1 inhibitor) were purchased from MedChem Express (Monmouth Junction, NJ, USA). KC7F2, gefitinib, and cisplatin were obtained from APExBIO (Houston, TX, USA). Chloroquine (CQ) was acquired from Sigma (St. Louis, MO, USA). Rapamycin was obtained from Selleck Chemicals (Houston, TX, USA). Cell Counting Kit-8 (CCK-8) was purchased from Beyotime Biotech (Shanghai, China).