Caco 2

Human CRC cell lines SW480, SW620, Caco-2 and HCT116 were purchased from the Cancer Institute of the Chinese Academy of Medical Sciences. SW480, SW620 and Caco-2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Biological Industries, Israel). HCT116 was cultured in McCoy’s 5A Medium (Biological Industries, Israel). All cells were supplemented with 10% fetal bovine serum (FBS, Biological Industries, Israel) and 1% penicillin–streptomycin (Biological Industries, Israel) and cultured in 5%CO₂ incubator at 37 °C. Small interfering RNA (siRNA) against COLEC12 (si-COLEC12) and the corresponding negative control (NC) were synthesized by GenePharma (Jiangsu, China), and sequences of siRNA were as follows: si-COLEC12-1 (sense 5′–3′): GCAAUCUGCAGAACCAAAUTT; si-COLEC12-2 (sense 5′–3′): GCGAAUCAAGAACGACUUUTT; si-COLEC12-3 (sense 5′–3′): GCUGACCAGCAAUCUAAAUTT; si-NC (sense 5′–3′):UUCUCCGAACGUGUCACGUTT. Following the manufacturer’s instructions, CRC cells cultured in 6-well plates were transfected at 70%–80% confluence using Lipofectamine 3000 (Invitrogen, USA), and the knockdown efficiency of COLEC12 protein expression was measured with goat anti-human COLEC12 polyclonal antibody (R&D Systems, USA) by western blotting (WB) after 48 h of cultivation.

The human embryonic kidney cell line HEK-293T (ATCC CRL-3216) and CRC cell lines HCT116 (ATCC CCL-247), HT29 (ATCC HTB-38), SW480 (ATCC CCL-228), and SW620 (ATCC CCL-227) were purchased from American Type Culture Collection (ATCC; http://www.atcc.org/). HCT116/L-OHP cells with oxaliplatin resistance were purchased from MEIXUAN Biological Science & Technology (Shanghai, China). The CRC cell lines HCT8, Caco2, RKO and LOVO were kindly provided by Professor Wancai Yang (Key Laboratory of Precision Oncology of Shandong Higher Education, Institute of Precision Medicine, Jining Medical University). The normal colon epithelial cell line 8401 (also known as CCD 841 CoN, ATCC® CRL­ 1790™) was kindly provided by Professor Lunquan Sun (Xiangya Hospital, Central South University, Changsha, China). HCT116 cells were maintained in McCoy’s 5 A medium (Biological Industries, Kibbutz Belt HaEmek, Israel) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Kibbutz Belt HaEmek, Israel). The oxaliplatin-resistant cell line HCT116/L-CHOP was maintained in RPMI 1640 medium supplemented with 10000 ng/ml oxaliplatin (Selleck, Selleck Chemicals, USA). HEK-293T, HT29, SW480, SW620, HCT8, Caco2, RKO and LOVO cells were maintained in RPMI 1640 (Biological Industries, Kibbutz Belt HaEmek, Israel) supplemented with 10% FBS. All cells were cultured at 37 °C in the presence of 5% CO₂.

The normal human colon mucosal epithelial cell line NCM460 and human colorectal cell lines HCT116, SW480, and SW620 used in this study were purchased from the American Type Culture Collection (ATCC; ⁸). The colorectal cell lines HCT8, Caco2, and RKO were kindly provided by Professor Wancai Yang (Key Laboratory of Precision Oncology of Shandong Higher Education, Institute of Precision Medicine, Jining Medical University). The NCM460, SW480, Caco2, HCT116, HCT8, SW620, and RKO cells were maintained in RPMI-1640 (Biological industries) with 10% FBS. All cells were cultured at 37°C with 5% CO₂.