5 fluorouracil

AGS, NCI-N87, and MKN-45 cells were obtained from the America Type Culture Collection (ATCC). SNU-668 cells were obtained from the Korean Cell Line Bank (KCLB). AGS, MKN-45 and SNU-668 cells were maintained in RPMI 1640. NCI-N87 gastric cancer cell line was maintained in DMEM/F12. All media were supplemented with 10% FBS, 100 U/mL penicillin and 100 mg/mL streptomycin, and L-glutamine 2 mmol/L (“regular media”). Cancer cell lines were actively passaged for less than 6 months from the time that they were received from ATCC and United Kingdom Coordinating Committee on Cancer Research (UKCCCR) guidelines were followed (41 (link)). 5-fluorouracil was purchased from US Biological, and cisplatin was purchased from Enzo Life Sciences. PI3K inhibitor (LY294002), JNK inhibitor (SP600125), ERK inhibitor (U0126) and p38 MAP kinase inhibitor SB203580 were purchased from Calbiochem. Rac1 inhibitor (NSC23766) was purchased from Santa Cruz Biotechnology. 5-fluorouracil was purchased from US Biological, and cisplatin was purchased from Enzo Life Sciences. Gastric cancer cell lines were grown as spheroids and spheroids were counted as previously described (14 ).

SNU-638, SNU-719, AGS, and NCI-N87 (subsequently referred to as N87) are Lauren intestinal-type gastric adenocarcinoma cell lines, and KATOIII, SNU-668, SNU-601, and MKN-45 are Lauren diffuse-type gastric adenocarcinoma cell lines. AGS, N87, SNU-601, and MKN-45 cells were obtained from the America Type Culture Collection (ATCC). SNU-638, SNU-719, KATOIII, and SNU-601 are from the Korean Cell Line Bank (KCLB). KATOIII cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM). All other gastric cancer cell lines were maintained in RPMI 1640. All media were supplemented with 10% FBS, 100 U/mL penicillin and 100 mg/mL streptomycin, and L-glutamine 2 mmol/L (“regular media”). Cancer cell lines were actively passaged for less than 6 months from the time that they were received from ATCC, and UKCCCR guidelines were followed (13 (link)). 5-fluorouracil was purchased from US Biological, and cisplatin was purchased from Enzo Life Sciences. RhoA inhibitor (Rhosin), PI3K inhibitor (LY294002), JNK inhibitor (SP600125), MEK I and II inhibitor (U0126) and p38 MAP kinase inhibitor (SB202190) were purchased from Calbiochem. ROCK inhibitor (Fasudil) was purchased from Abcam.

AGS and MKN-45 are Lauren diffuse-type gastric adenocarcinoma cell lines, and NCI-N87 (subsequently referred to as N87) is a Lauren intestinal-type gastric adenocarcinoma cell line. These cells were obtained from the America Type Culture Collection (ATCC, Manassas, VA). CFPAC-1 and PANC-1 are pancreatic adenocarcinoma cell lines and were also obtained from ATCC. Cell lines were maintained in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/ml streptomycin, and L-glutamine 2 mM (“regular media”). Cancer cell lines were actively passaged for less than 6 months from the time that they were received from ATCC, and UKCCCR guidelines were followed (14 (link)). Vismodegib was purchased from LC Laboratories (Woburn, MA). 5-fluorouracil was purchased from US Biological. Cisplatin was purchased from Enzo Life Sciences (Farmingdale, NY).