Interaction of solute carrier family 39 member 1 and dynamin related protein 1 to facilitate development of hepatocellular carcinoma by impairing mitochondrial quality control

In this study, HCC samples from 12 non-relapsed and 15 relapsed HCC patients after surgically resection, were collected for RNA sequencing. Liver-specific SLC39A1 knockout mice (SLC39A1lko) were generated by crossing Alb-Cre mice and SLC39A1flox/flox mice. Liver samples from mice were analyzed for inflammation, fibrosis and proliferation. Mitochondrial dynamics, autophagy, ROS and mitochondrial membrane potential (MMP), were detected. Co-immunoprecipitation and molecular docking were used to identify protein interactions. Targeting 1-28 peptide of SLC39A1 was designed. Lentiviral pLKO.1-shRNA plasmids targeting SLC39A1, SLC50A1 and SLC66A3. For lentivirus packaging, pLKO.1-control, pLKO.1-shSLC39A1, pLKO.1-shSLC50A1 and pLKO.1-shSLC66A3 were transfected into HEK-293T cells using a co-transfection system (pMD2G: psPAX2: target plasmids = 1:3:4) with PEI. The culture supernatant was collected at 48 and 72 h after transfection and then filtered through a 450 nm filter. When cancer cells reached approximately 30% coverage in 6-well plates, 0.5ml of lentivirus, 0.5 ml of fresh medium were added for infection. After incubation for 24 h, puromycin (2 µg/µl) was used for three days to select cells that were successfully infected.