Hct 116

The human malignant growth cancer cell lines MDA-MB-231, MCF-7, Skbr3 (breast cancer), HeLa (cervical cancer), HT-29, HCT 116 (colorectal adenocarcinoma) and normal cell line chang liver are procured from the National Centre for Cell Science, Pune, India. The cells were developed in Dulbecco’s Modified Eagle Medium (DMEM) medium enhanced with 10% inactivated Fetal Bovine Serum (FBS), 100 μg of streptomycin/ml, 100 U/ml of penicillin and incubated at 37 ± 2°C with 5% CO₂. MTT dye (3-(4, 5-dimethylthiazol-2-yl) - 2, 5-diphenyl tetrazolium bromide) was acquired from Sigma Aldrich (Mumbai, India). All the experimental reagents and solvents were purchased from sigma Aldrich chemicals Pvt Ltd. Recombinant VEGF₁₆₅ (Cell signalling technology), P-p44/42 MAPK, P-SAPK/JNK (G9) and JNK antibodies were purchased from cell signaling technology, USA. HRP-conjugated mouse secondary antibody and HRP-conjugated goat anti-rabbit secondary antibodies were purchased from cell signalling technology. Actin antibody was obtained from Abcam, USA.

Colorectal cancer cell lines (HCT-116 and Caco-2) and control cell line (HEK-293) were procured from the National Centre For Cell Science (NCCS), Pune, India. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) added with 10% heat-inactivated fetal bovine serum (FBS), 1% of l-glutamine, and 1% of antibiotics (penicillin/streptomycin) and incubated at 37°C in a humidified atmosphere (5% CO₂).

The human cancer cell lines MDA-MB-231, 143B, AGS, MCF7, T47D, HCT116, HT29, H23, H460, and H522 cell lines were obtained from the National Centre for Cell Sciences (NCCS), Pune (India). The cells have been authenticated by the NCCS using STR. H1299 cells were purchased from ATCC, United States. Cell were cultured in recommended cell culture media supplemented with fetal bovine serum (10% v/v) and 100 U/mL Penicillin and 100 μg/ml Streptomycin. Cells were grown in a humidified incubator at 37°C in 5% CO₂/95% atmospheric air. All the cell lines were tested for mycoplasma contamination, and experiments were performed in mycoplasma-free cells only.

HeLa (human cervical cancer), HCT116 (human colon cancer), MEF (mouse embryonic fibroblast) and HEK 293T (human embryonic kidney epithelial cell line) were purchased from National Centre for Cell Science, Pune, India. Nalm6 and Reh cells were from Dr M. R. Lieber (USA), and Rho(0) cells were from J. Neuzil (Australia). The identity of all these cell lines was confirmed by performing STR profiling (DNA labs India, Hyderabad, India). All the cell lines were found to be free of mycoplasma contamination. The Mycoplasma test was conducted in the laboratory using the MYCOseq kit (Thermo Fisher Scientific, Waltham, Massachusetts) and further validated by DNA labs India, Hyderabad, India. Cells were cultured in RPMI1640 or MEM medium supplemented with 10–15% fetal bovine serum (FBS), 100 μg/ml Penicillin, and 100 μg/ml streptomycin and incubated at 37 °C in a humidified atmosphere containing 5% CO₂ as described before (Ghosh et al., 2022 (link); Kumari et al., 2021 (link)). Rho(0) cells of B16 cells origin were cultured in DMEM medium supplemented with 15% fetal bovine serum (FBS), 100 μg/ml Penicillin, 100 μg/ml streptomycin, 50 mg/ml Uridine and 1 mM Sodium pyruvate and incubated as described above (Dong et al., 2017 (link); Tan et al., 2015 (link)).