Pm150910

Human fetal lung fibroblast (HFL1, Cat. No. SCSP5049) was purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China) and cultured in Ham’s F-12K (Procell, PM150910) medium supplemented with 10% fetal bovine serum (FBS) (Gibco, 10099141C) and 1% penicillin-streptomycin Solution (Hyclone, SV30010) in an incubator at 37°C with 5% CO₂ atmosphere.

A549 and A549/DDP cells were purchased from Procell (Wuhan, China) and cultured at 37°C with 5% CO₂ in Ham's F-12K (PM150910, Procell) containing 10% FBS (164210-50, Procell) and 1% Penicillin-Streptomycin Solution (PB180120, Procell). Besides, H1299 and H1299/DDP (Biovector, Beijing, China) were cultured in RPMI-1640 medium plus 10% FBS and 1% Penicillin-Streptomycin Solution. In functional experiments, A549/DDP and H1299/DDP cells were treated with 1 μM anlotinib (CTTQ-PHARMA, Jiangsu, China), 2 μM DDP (Sigma-Aldrich, St. Louis, MO, USA), or 1 μM anlotinib+2 μM DDP for 24 h.

Lung adenocarcinoma cell lines including A549 (CM-0016) and H1299 (CM-0165) were purchased from Procell (Wuhan, China). Both cell lines were authenticated by short tandem repeat (STR) profiling. A549 cells were cultured in Ham’s F-12K medium (PM150910, Procell) supplemented with 10% FBS (164210-500, Procell) and 1% P/S (PB180120, Procell). H1299 cells were cultured in RPMI-1640 medium (PM150110, Procell) plus 10% FBS (164210-500, Procell) and 1% P/S(PB180120, Procell). All cell lines were routinely maintained at 37 °C in a 5% CO₂ incubator.

Lung cancer cells (A549 and SPC-A-1) were purchased from Procell (Wuhan, China) or Otwo Biotech (Guangzhou, China), and cultured in Ham's F-12 K (PM150910, Procell, Wuhan, China) or RPMI-1640 medium (PM150110, Procell, Wuhan, China) supplemented with 10% FBS (164,210, Procell, Wuhan, China) and 1% Penicillin–Streptomycin (PB180120, Procell, Wuhan, China).

The murine intestinal epithelial cell line MODE-K (Shanghai Jining Industrial Co., Ltd., Shanghai, China) was adopted for the study. The cells were washed with a cell culture solution that was comprised of the 1640 medium (PM150910; Procell, Wuhan, China) +10% fetal bovine serum (FBS, Gibco, Carlsbad, NM, United States) solution (pH 7.2–7.4) +1% penicillin-streptomycin (Sigma, St. Louis, MO, United States) to remove DMSO. The resuscitated cells were cultured in a fresh cell culture solution and under 5% CO₂ saturated humidity at 37°C. The cell culture experiments were performed in triplicate with α-chaconine treatment at concentrations of 0, 0.4, and 0.8 μg/mL. α-Chaconine stock solution (16 mg/100 mL) was made and diluted with the cell culture solution. The choice of the concentrations was based on pilot trials. Measurements were performed in triplicate after incubation for 24, 48, and 72 h.