Hct116

Human COAD cell lines (SW480, HT29, and HCT116) and a human colon epithelial cell line (NCM460) were purchased from Procell Life Science & Technology Co. NCM460 cells were cultured in 90% Dulbecco's Modified Eagle Medium: F12; SW480 cells were cultured in 90% L-15 base medium; and HT29 and HCT116 cells were cultured in 90%McCOY's 5 A base medium. The cells were supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution at 37°C in 95% air and 5% CO₂, which was then replaced with a complete medium, and the cells were cultured for 24–48 h. The cells were then transfected with hsa-miR-135b-5p inhibitors or a negative control using Lipofectamine 2000 following the manufacturer's instructions (incubation at room temperature for 20 min).

Two human CRC cell lines, HCT116 and SW480, were obtained from Procell (Wuhan, China) and the National Infrastructure of Cell Line Resource (NICR), respectively. HCT116 and SW480 cells were cultured in McCoy 5A (Procell, China) and IMDM (Procell, China) medium containing 10% FBS, respectively. Cells were maintained in a wet incubator with 5% CO₂ at 37°C. The recombinant lentivirus used for SH3TC2 knockdown (sh-SH3TC2#1, #2) and the corresponding control lentivirus (sh-NC) were purchased from GeneChem (Shanghai, China). The overexpression plasmids for wide-type (OE-YTHDF1-WT) and mutant (OE-YTHDF1-Mut) YTHDF1 were also obtained from GeneChem (Shanghai, China). The reagents HiTransG A (GeneChem, China) and Lipo3000 (Thermo, USA) were utilized to transfect lentiviruses and plasmids into CRC cells, respectively.

Human CRC cell lines (HT-29, LoVo, SW480 and HCT116) and human umbilical vein endothelial cells (HUVECs) were purchased from Procell Life Science &Technology (Wuhan, China). The normal Human Colonic Epithelial Cells were obtained from Zhong Qiao Xin Zhou Biotechnology (Shanghai, China). HT-29 and HCT116 cells were cultured in McCoy’s 5A medium (PM150710, Procell, Wuhan) with 10% fetal bovine serum (FBS, 04–011-1A, BI) in an incubator at 37°C in 5% CO_2. SW480 cells were cultured in L-15 medium (PM151010, Procell, Wuhan) supplemented with 10% FBS in an incubator at 37°C in 5% CO_2°C LoVo cells were cultured in Ham’s F-12 K (PM150910, Procell, Wuhan) medium containing 10% FBS in an incubator at 37°C in 5% CO_2. For cell passage, HUVECs were normally cultured in Ham’s F-12 K (PM150910, Procell, Wuhan) medium containing 10% FBS in an incubator at 37°C in 5% CO₂.

All cell experiments were performed using sterile tools. The HEK293 cell line and human colorectal carcinoma cell HCT116 and DLD1 cell lines were purchased from Procell Life Science & Technology (Procell, Wuhan, China). All cells were cultured in a Thermo incubator at 37°C in 5% CO2, with each line in a different type of medium. The HCT116 cells were cultured in McCoy’s 5A medium (Procell, Wuhan, China) with 10% fetal bovine serum (FBS) (Procell, Wuhan, China). The DLD1 cells were cultured in RPMI-1640 medium (Procell, Wuhan, China), and Dulbecco’s Modified Eagle Medium (Procell, Wuhan, China) was used for the HEK293 cells.