Sw620

Human CRC cell lines SW480 (BeNa Culture Collection, Xinyang, China), SW620 (Procell Life Science & Technology Co., Ltd., Wuhan, China), and HT-29 (Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, China) were cultured in RPMI 1640 Medium (Livning Biotechnology Co., Ltd., Beijing, China), LoVo cells (Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, China) were cultured in DMEM/F12 (Livning Biotechnology Co., Ltd., Beijing, China), and normal human liver cell line QSG-7701 (Beyotime Institute of Biotechnology, Shanghai, China) was cultured in DMEM/high glucose (Livning Biotechnology Co., Ltd., Beijing, China) containing 10% fetal bovine serum (FBS) (Gibco Life Technologies, San Jose, CA, USA). The cells were cultured in a constant temperature incubator at 37 °C, 5% CO₂.

The CRC cell lines-HCT116, SW480, SW620 and normal cell line-NCM460 were bought from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and short tandem repeat (STR) profiling was applied to confirm. All the cell lines were cultured in DMEM (Gibco) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin supplied. Then, the cell lines were grown with 5% CO₂ at 37 °C in humidified air.

Colon cancer cells (SW620, SW480, Caco-2, and LoVo) and 293 T were purchased from Procell (Wuhan, China). HT29 was purchased from Zhongqiao xinzhou Biological Technology Co., Ltd (Shanghai, China). Caco-2 was cultured in Minimum Essential Medium (Gibco, USA) supplemented with 10% fetal bovine serum (Hyclone, USA). LoVo was cultured in Ham’s F12 K Medium (Procell, China) containing 10% fetal bovine serum. SW620 and SW480 were cultured in Roswell Park Memorial Institute 1640 Medium (Gibco, USA) supplemented with 10% fetal bovine serum. HT29 and 293 T were cultured in Dulbecco’s Modified Eagle’s medium (Gibco, USA) supplemented with 10% fetal bovine serum. All cells were cultured in an atmosphere at 37 °C with 5% CO₂.
DNAJC3-AS1 siRNA (si-DNAJC3-AS1), and negative control siRNA (si-NC) were synthesized by JTS scientific (Wuhan, China). miR-214-3p mimics, miR-214-3p inhibitors and negative controls were obtained from JTS scientific (Wuhan, China). Colon cancer cells were transfected using the Lipofectamine 2000 transfection reagent (Invitrogen, USA) according to the manufacturer’s protocol.

Human embryonic kidney cell line 293T (CL-0005), human CC cell lines SW480 (CL-0223B), HCT116 (CL-0096), RKO (CL-0196), HT29 (CL-0118), and SW620 (CL-0225B) were provided by Procell (Wuhan, China). Human normal colonic epithelial cells FHC and murine CC cells MC38 were kindly gifted from Dr. Jikun Li (Shanghai Jiao Tong University School of Medicine). Cells were cultured in DMEM supplemented with 10% FBS and antibiotics, at 37 °C and 5% CO₂. The tumor cell lines devoid of mtDNA (ρ0) were established and cultured as described previously^{40 (link)}. Briefly, parental cells were grown in a medium supplemented with 100 ng mL⁻¹ EtBr, 50 µg mL⁻¹ uridine, and 1 mM pyruvate for 3 months, followed by verification of mtDNA depletion using qPCR and electron microscopy. All cell lines were authenticated by STR profiling and routinely tested negative for mycoplasma contamination.

CRC cell lines NCM460, RKO, HCT116, SW480, Colo320, DLD-1, HCE8693, SW620, and HT29, and the normal human colon epithelial cell line FHC were purchased from Wuhan Procell and ATCC. NCM460, DLD-1, HCE8693, and Colo320 were maintained in RPMI-1640, RKO in MEM, HCT116 and HT29 in McCoy’s 5A, SW620, SW480 in Leibovitz’s L-15 medium that supplied with 10% FBS (BI, Israel) and 1% penicillin and streptomycin (Sigma, USA) and maintained in 37° C incubator with 5% CO₂.
METTL16 proteins siRNAs and overexpression vectors were obtained from GenePharma (Shanghai, China). Cells were transfected with indicated oligonucleotides using the Lipofectamine 2000 reagent (Thermo, USA) as per manufacturer’s protocol. Transfection efficacy was examined 2 days after transfection.

The present study was approved by the ethics committee of the Affiliated Cancer Hospital and Institute of Guangzhou Medical University. The colorectal cancer PDOs were derived from patients in the Affiliated Cancer Hospital and Institute of Guangzhou Medical University, between May and August 2020. The patients with colorectal cancer were diagnosed on the basis of pathology results, treated with cytoreductive surgery, and followed up with or without hyperthermic intraperitoneal chemotherapy. All colorectal cancer cell lines used in this study, including SW620, SW480, DLD-1, and COLO 205, were purchased from Procell Life Science & Technology Co. Ltd.

The pathology specimens were acquired from the biospecimen repository of Xinqiao Hospital. The 93 patients from our Department of General Surgery, Xinqiao Hospital, all underwent laparoscopic left (or right) hemicolectomy during 2015–2016. All patients signed an informed consent upon admission to our hospital. The application for surgical specimens was approved by the Clinical Ethics Committee of the Second Affiliated Hospital of Army Medical University of the PLA (no. 2022-036-01). The study methodologies strictly conformed to the standards set by the Declaration of Helsinki.
Colon cancer cell lines, including SW620 and HCT116, were purchased from Procell Life Science & Technology Co., Ltd. Then, they were cultured in the corresponding complete medium, which was Leibovitz’s L-15 (PM151010)+10% FBS (164210-500)+1% P/S (PB180120) or MEM+10% FBS (164210-500)+1% P/S (PB180120), in an incubator of 37°C constant temperature with 5% CO₂.