Ht 29

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.

The human colorectal adenocarcinoma cell lines (Caco-2 and HT-29) were purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd. and maintained in DMEM medium (BasalMedia, Shanghai, China) supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, United States) and incubated at 5% CO₂ at 37°C.
The MMP9 sequence was synthesized by General Biol and then inserted into the pCDH-CMV-MCS-EF1-copGFP-T2A-Puro lentiviral expression vector plasmids (General Biol, Anhui, China), called MMP9 OE plasmids. In addition, lentivirus-containing shRNA targeting MMP9 (MMP9 shRNA1: 5′-GGA​ATA​CCT​GTA​CCG​CTA​TGG-3′, MMP9 shRNA2: 5′- GCA​GAC​ATC​GTC​ATC​CAG​TTT​C-3′ and MMP9 shRNA3: 5′-GCT​TAG​ATC​ATT​CCT​CAG​TGC-3′) were synthesized by GenePharma (Shanghai, China). After that, 293T cells were transfected with indicated lentiviral plasmids, packaging plasmids (pAX2) and envelope plasmid (pMD2. G) for 72 h. Next, virus-containing supernatants was filtered through a filter membrane (0.22 μm pore size), and then transduced into Caco-2 and HT-29 cells. Later on, the infected cells were selected by puromycin.