Flotillin-1 promotes the progression of hepatocellular carcinoma by activating TFE3-mediated Golgi stress response via inhibition of mTORC1/2

Figure 2 Flotillin-1 promotes proliferation, invasion, and migration of hepatocellular carcinoma cells in vitro and in vivo. A: Expressions of flotillin-1 (FLOT1) protein in normal hepatocyte (L-02) and hepatocellular carcinoma lines (SNU-387, Hep-G2, SNU-398, and Huh-7) were detected by western blot; B: Quantitative reverse transcription polymerase chain reaction quantification of FLOT1 mRNA. n = 3, ^aP < 0.05 vs L-02, ^bP < 0.01 vs L-02; C-F: FLOT1 overexpression was transfected in Huh-7 cells and FLOT1 short-hairpin RNA was transfected in SNU-387 cells (n = 3). CCK-8 assay was used to detect the viability of Huh-7 and SNU-387 cells at 24, 48, and 72 hours (C). Cell cycle was measured by flow cytometry (D). Cell invasion and migration ability of Huh-7 and SNU-387 cells (E). Apoptosis-related proteins [cleaved-caspase 3 and cleaved-poly(ADP-ribose) polymerase] were detected by western blot (F); G: Xenograft tumor picture (n = 5) of Huh-7 cells with overexpression and SNU-387 cells with knockdown (short-hairpin RNA-FLOT1) in the Balb/c nude mice; H: Tumor volumes of Huh-7 and SNU-387 cells in the nude mice for 24 days (n = 6); I: Ki-67 and FLOT1 expressions in the tumor tissue of Balb/c nude mice transplanted with Huh-7 and SNU-387 cells were detected by immunohistochemical staining (n = 3). Scale bar = 50 μm. ^aP < 0.05, ^bP < 0.01. Values are expressed as mean ± SD, and comparisons between two groups were performed using the t-test. FLOT1: Flotillin-1; OV: Overexpression; PARP: Poly(ADP-ribose) polymerase.