Knockdown of EFEMP1 Promotes Ferroptosis by Inactivating PI3K/AKT to overcome the Resistance of Hepatocellular Carcinoma Cells to Sorafenib.
We found epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) is up-regulated in liver cancer cells exposed to sorafenib for a long time using a bioinformatic tool. Here, the mechanism of EFEMP1 in sorafenib resistance of hepatocellular carcinoma (HCC) cells was explored.
Human HCC cell lines Huh7 and HCCLM3 received low concentrations of sorafenib for a long time to construct sorafenib-resistant Huh7 and HCCLM3 (Huh7-SR and HCCLM3-SR) cells. HCCLM3, HCCLM3-SR, Huh7 and Huh7-SR cells received sorafenib, and the cell viability was assayed by CCK-8 method. HCCLM3-SR and Huh7-SR cells were transfected before sorafenib treatment, and these cells apoptosis was determined with flow cytometry assay. Ferroptosis-related index, EFEMP1, phosphorylated PI3K (p-PI3K), p-AKT level in HCCLM3, HCCLM3-SR, Huh7 and Huh7-SR cells was detected using flow cytometry assay, colorimetry, qRT-PCR and Western blot analysis, respectively.
Following sorafenib treatment, HCCLM3-SR (8/16 μM) and Huh7-SR (4/8/16/32 μM) cell viability was higher than HCCLM3 and Huh7 cell viability. HCCLM3-SR and Huh7-SR cells presented higher IC50 of sorafenib. Following sorafenib (7 μM) treatment, ROS, MDA, TBARS, Fe2+ level in HCCLM3-SR and Huh7-SR cells was lower, and SLC7A11, GPX4, EFEMP1, p-AKT and p-PI3K level in Huh7-SR and HCCLM3-SR cells was higher than those in HCCLM3 and Huh7 cells. Under sorafenib (7 μM) treatment, EFEMP1 silencing promoted apoptosis, up-regulated ROS, MDA, TBARS, Fe2+ level and inhibited SLC7A11, GPX4, p-AKT and p-PI3K expression in Huh7-SR and HCCLM3-SR cells.
Knockdown of EFEMP1 promotes ferroptosis by inactivating PI3K/AKT to resensitize sorafenib-resistant HCC cells to sorafenib.
Human HCC cell lines Huh7 and HCCLM3 received low concentrations of sorafenib for a long time to construct sorafenib-resistant Huh7 and HCCLM3 (Huh7-SR and HCCLM3-SR) cells. HCCLM3, HCCLM3-SR, Huh7 and Huh7-SR cells received sorafenib, and the cell viability was assayed by CCK-8 method. HCCLM3-SR and Huh7-SR cells were transfected before sorafenib treatment, and these cells apoptosis was determined with flow cytometry assay. Ferroptosis-related index, EFEMP1, phosphorylated PI3K (p-PI3K), p-AKT level in HCCLM3, HCCLM3-SR, Huh7 and Huh7-SR cells was detected using flow cytometry assay, colorimetry, qRT-PCR and Western blot analysis, respectively.
Following sorafenib treatment, HCCLM3-SR (8/16 μM) and Huh7-SR (4/8/16/32 μM) cell viability was higher than HCCLM3 and Huh7 cell viability. HCCLM3-SR and Huh7-SR cells presented higher IC50 of sorafenib. Following sorafenib (7 μM) treatment, ROS, MDA, TBARS, Fe2+ level in HCCLM3-SR and Huh7-SR cells was lower, and SLC7A11, GPX4, EFEMP1, p-AKT and p-PI3K level in Huh7-SR and HCCLM3-SR cells was higher than those in HCCLM3 and Huh7 cells. Under sorafenib (7 μM) treatment, EFEMP1 silencing promoted apoptosis, up-regulated ROS, MDA, TBARS, Fe2+ level and inhibited SLC7A11, GPX4, p-AKT and p-PI3K expression in Huh7-SR and HCCLM3-SR cells.
Knockdown of EFEMP1 promotes ferroptosis by inactivating PI3K/AKT to resensitize sorafenib-resistant HCC cells to sorafenib.