Mkn 45

The AGS, MKN45, SGC7901, and HGC27 GC cell lines and the control GES-1 gastric mucosal cell line were obtained from GeneChem (Shanghai, China). The MFC cell were obtained from Pricella (Wuhan, China). All cells were grown in RPMI-1640 supplemented with FBS (10%) at 37 °C in a humidified 5% CO₂ incubator.

Seven GC cell lines (AGS, BGC823, MGC803, MKN45, MKN1, SGC7901, and HGC27), a gastric mucosa cell line (GES-1), and human kidney cell line (HEK293T) were purchased from GeneChem (Shanghai, China). Cells were cultured in RPMI-1640 medium supplemented with 10% FBS at 37 °C in a humidified atmosphere with 5% CO₂.
DAPI, Baf-A1, and 3-MA were obtained from Solarbio (Beijing, China). NAC was purchased from Beyotime (Shanghai, China), and 740Y-P (also known as 740YPDGFR) was obtained from MCE (Monmouth Junction, NJ, USA).

Gastric cancer cell lines: HGC-27, SNU-1, SGC-7901, NCI-N87, KATOIII, AGS, MKN-28, MKN-45, BGC-823, MGC-803 and GES-1 were purchased from the Chinese Academy of Sciences (Shanghai, China), and were cultured in corresponding medium containing 10% fetal bovine serum (FBS). All the cell lines were kept at 37°C in 5% CO₂. To obtain KLF2 over-expressing cells, BGC823 and MKN45 cells were infected with lentivirus encoding GFP-KLF2 (KLF2) or GFP (as control) (Shanghai Genechem Co., LTD.). MGC803 cells were infected with lentivirus encoding shRNA KLF2 or scrambled shRNA used as negative Control Duplex (Hanbio Biotechnology Co., Ltd.) to obtain KLF2 knockdown cells. Three different shRNA sequences were used: shRNA1: sense5’-GATCCGCGGCACCGACGACGACCTCAATTCAAGAGATTGAGGTCGTCGTCGGTGCCGTTTTTTC-3’, antisense 5’- AATTGAAAAAACGGCACCGACGACGACCTCAATCTCTTGAATTGAGGTCGTCGTCGGTGCCGCG-3’; shRNA2: sense5’- GATCCGCCTACACCAAGAGTTCGCATCTGAATTCAAGAGATTCAGATGCGAACTCTTGGTGTAGGTTTTTTC-3’, antisense 5’- AATTGAAAAAACCTACACCAAGAGTTCGCATCTGAATCTCTTGAATTCAGATGCGAACTCTTGGTGTAGGCG-3’; shRNA3: 5’- GATCCGCGCTGCACATGAAACGGCACATGTATTCAAGAGATACATGTGCCGTTTCATGTGCAGCGTTTTTTC-3’, antisense 5’- AATTGAAAAAACGCTGCACATGAAACGGCACATGTATCTCTTGAATACATGTGCCGTTTCATGTGCAGCGCG-3’. The transfected cells were selected in culture medium plus puromycin (1.0 μg/ml) for 3 weeks to obtain stable KLF2-overexpressing and KLF2-deficient cells.

Tumor tissues and adjacent non-tumor tissues were randomly collected from 138 GC patients treated between April 2012 and October 2014 at the Department of Oncology Surgery, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University, PR China. None of the patients reported to have been pretreated with chemotherapy or radiotherapy. By reviewing the patients’ pathology records, we obtained their clinicopathological such as their age and gender, histology data, lymph node metastasis status, lymphatic and venous invasion status, tumor size, T stage and pTNM stage. Tumor stage was defined based on the criteria of the American Joint Committee on Cancer (AJCC). The study was approved by the Ethical Committee of Xi'an Jiaotong University. Informed consent was obtained from all the patients before samples were collected. Human GC cell lines, including BGC-823, AGS, MKN-45, SGC-7901 and GES-1, were obtained from the Cell Bank (Shanghai Genechem Co., Ltd, Shanghai, China). The cell lines have been authenticated and tested by the Cell Bank. For verification, we performed mycoplasma tests in our laboratory, and the cell behavior and morphology were proved consistent with the descriptions in the Cell Bank. Cells (1 × 10⁵ cells/ml) were cultured in RPMI-1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco) at 37 °C.