Colo205

The human CRC cell lines HCT15, HCT116, SW620, LS174t, LoVo, HCT8, Colo‐205, HT29, and the FHC cell were purchased from the American Type Culture Collection (ATCC). The M5 cell, the ability of liver metastasis of which was enhanced, was obtained by the in vivo selection of SW480 from our previous study.⁵² The FHC cell and HUVECs were purchased from the cell bank at the Chinese Academy of Sciences (Shanghai, China). FHC, LS174t, SW620, HCT116, HCT8, LoVo, Colo‐205, HT29, HCT15, and HUVECs cells were cultured in RPMI 1640 or DMEM medium (Gibco) with 10% fetal bovine serum (Gibco BRL). They were maintained at 37°C in a humidified 5% CO₂ atmosphere.

The human colon adenocarcinoma HT29, SW620, SW480, HCT15, LoVo, 293T, and 293FT cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). KM12 was from Japan and Colo-205 was from Shanghai Institute of Biochemistry and Cell Biology. HT29 and HCT116 cells were grown in McCoy's 5A medium (Sigma) supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY). SW620 and SW480 cells were grown in L-15 (Gibco, Grand Island, NY) supplemented with 10% FBS. HCT15 and Colo-205 were grown in RPMI-1640 (Gibco, Grand Island, NY) supplemented with 4.5 g/L glucose, 0.11 g/L sodium pyruvate, and 10% FBS. LoVo was grown in F-12 (Gibco, Grand Island, NY) supplemented with 0.29 g/L glutamine and 10% FBS. KM12 was grown in RPMI-1640 (Gibco, Grand Island, NY) supplemented with 10% FBS. The 293FT Lentiviral Expression System and 293T were cultured in Delbecco's modified Eagle medium (Gibco, Grand Island, NY) supplemented with 10% FBS.

COLO-205, HT-29 and LoVo purchased from ATCC were used in this study. These cells have performed STR-PCR profile at MB Mission Biotech. The cell was grown in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and kanamycine (100 ng/mL) in a humidified incubator (37 °C, 5% CO2). For transient transfection of the indicated constructs into COLO-205, jetPEI-HUVEC transfection reagent (Polyplus Transfection, Bioparc, France) was used and the transfection was performed according to the manufacturer’s protocol. Briefly, jetPEI-COLO-205/DNA mixture was added drop-wise into the DMEM+GlutamaxTM I medium (GIBCO) containing 2% FBS, mixed gently, and incubated in a humidified 37 °C incubator for 4 h. The growth medium was then replaced and the cell was incubated for the next 24 h.

Authenticated CRC lines were obtained from ECACC: HT29 (ECACC 91072201), Colo205 (ECACC 87061208), Colo320DM (ECACC 87061205), SW480 (ECACC 87092801), and SW620 (ECACC 87051203). Authenticated primary human colonocytes were obtained from ATCC: CCD-18Co (ATCC CRL-1459) and CCD-841 CoN (ATCC CRL-1790). All lines were regularly monitored for mycoplasma contamination every 2 months by PCR^{40 (link)}, and only mycoplasma-free cells were used for studies. All cells were used for no more than 15 passages. All cells were grown at 37 °C in HEPA-filtered humidified air in 5% CO₂, in media supplemented with 10% heat-inactivated FBS (Gibco, Paisley, Scotland, UK, Cat. no. 10099-141), L-Glu (2 mM, Lonza, Burton on Trent, UK, Cat. no. 17-606E) and penicillin/streptomycin (100 U/ml, Gibco, Cat. no. DE17-602E). Colo205 and Colo320DM cells were grown in RPMI medium (Gibco, Paisley, Scotland, UK, Cat. no. 31870), HT29 were grown in McCoy’s medium (Lonza, Burton on Trent, UK, Cat. no. 12-688F), SW480 and SW620 were grown in L15 medium (Lonza, Burton on Trent, UK, Cat. no. BE12-700F), and HCT-116 (17) were grown in DMEM (Gibco, Paisley, Scotland, UK, Cat. no. 21885-025). CCD-18Co and CCD-841 primary cultures were grown in EMEM (Sigma-Aldrich, Saint Louis, MO, USA, Cat. no. M2279).