Hct116

Two human colon cancer cell lines (HCT-116, SW-480) were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China). Cells were maintained in DMEM medium (SH30084.03, Hyclone, USA) containing 10% fetal bovine serum (11,885,084, Gibco, USA) in a humidified atmosphere with 5% CO₂ at 37 °C.

CC cells, including SW480, Colo-205, SW620, HCT15, DLD-1, HCT116, Caco-2, RKO and LOVO were bought from Shanghai Zhong Qiao Xin Zhou Biotechnology. SW480 and SW620 were cultured in a complete DMEM medium supplemented with 10% FBS (Gibco Grand Island, NY, USA). Colo-205, HCT15 and DLD-1 cells in a complete RPMI-1640 medium supplemented with 10% FBS. Caco-2 and RKO cells in a complete EMEM medium supplemented with 10% FBS. HCT116 cells in a complete Mccoy’s 5A medium supplemented with 10% FBS. LOVO cells in a complete F-12K medium supplemented with 10% FBS. All the experimental procedures described below were repeated thrice unless described otherwise.

HCT-116 and HT-29 cell lines were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co. Ltd. (Shanghai, China). The 293T cell line was provided by Stem Cell Bank, Chinese Academy of Sciences (Shanghai, China). Human colonic epithelial cells (HCECs) were kindly provided by the Traditional Chinese Medicine Hospital of Kunshan (Kuanshan, China). The HCECs, HCT-116 and 293T cells were cultured in DMEM-high glucose (HyClone, USA) supplemented with 10% foetal bovine serum (FBS; Gibco, South America) and 1% penicillin/streptomycin (P/S; Gibco, Australia). HT-29 cells were cultured in McCoy’s 5A (Gibco, Australia) with 10% FBS and 1% P/S.
Five colon cancer patients were random selected from the Department of Oncology, Kunshan Hospital of Traditional Chinese Medicine Affiliated with Nanjing University of Chinese Medicine. Patient demographic information is provided in Supplementary Table 6. From each patient, tissues were obtained from colonic tumours as well as from adjacent normal tissue (2 cm away from tumour). All protocols were approved by the Ethics Review Committee of Kunshan Hospital of Traditional Chinese Medicine (Approval No: KZY2017-16).

The human CRC cell lines HCT116, HT29 and human embryonic kidney 293T (293T) cells were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. The HCT116 and HT29 cell lines were cultured in McCoy's 5A Medium (Biological Industries), and the 293T in EMDM Medium supplemented with 10% fetal bovine serum (Biological Industries) at 37°C in 5% CO₂.

Human colon cancer cell lines HCT116 (non-metastatic colon cancer cells, ZQ0125), SW480 (moderately metastatic colon cancer cells, ZQ0063), Lovo (highly metastatic colon cancer cells, ZQ0058), and the mouse colon adenocarcinoma cell line MC38 (ZQ0933) were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co. Ltd (Shanghai, China). HCT116, SW480, and MC38 cells were cultured in DMEM (KGM12800-500, Keygen Biotech, Nanjing, China) with 10% fetal bovine serum (FBS, 164210-50, Procell, Wuhan, China). Lovo cells were cultured in Ham’s F-12K medium (G4560-500ML, Servicebio, Wuhan, China) supplemented with 10% FBS. All cells were maintained in a 5% CO₂-containing incubator (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C.

Human CRC cell line HCT116 was purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. Cells were cultured in DMEM medium containing 10% FBS with 1% penicillin-streptomycin solution (C0222, Beyotime, China) in an incubator at 37°C, 5% CO₂, and saturated humidity.
The silenced plasmid (NC), Lnc-S100B-2 silenced plasmid (sh-Lnc-S100B-2), Lnc-S100B-2 overexpressed plasmid (oe-Lnc-S100B-2), and MLLT10 silenced plasmid (sh-MLLT10) were purchased from HonorGene. Briefly, 5 μg plasmid was added to 250 μL serum-free medium and mixed. Lipofectamine 2000 (Invitrogen, USA) was used for transfection according to the manufacturer's instructions. The specific groups were as follows: a control group (without any treatment), an NC group (NC was transfected), a sh-Lnc-S100B-2 group (sh-Lnc-S100B-2 was transfected), a sh-MLLT10 group (sh-MLLT10 was transfected), and a sh-Lnc-S100B-2+sh-MLLT10 group (sh-Lnc-S100B-2 and sh-MLLT10 were transfected).