GATA5 inhibits hepatocellular carcinoma cells malignant behaviours by blocking expression of reprogramming genes

Abstract

Evidence indicated that GATA5 may suppress hepatocellular carcinoma (HCC) cell malignant transformation, but the mechanism of how GATA5 affects cancer cell reprogramming to inhibit HCC malignant behaviour is still unclear. In this study, we report that the expression of β‐catenin and reprogramming genes p‐Oct4, Nanog, Klf4, c‐myc and EpCAM was significantly higher in HCC tissues compared to normal liver tissues. In contrast, the expression of GATA5 was significantly lower in HCC tissues compared to normal liver tissues. Transfection of CDH‐GATA5 vectors into HCC cells (HLE, Bel 7402 and PLC/PRF/5 cells) increased the GATA5 expression and decreased the expression of β‐catenin and reprogramming genes p‐Oct4, Nanog, Klf4, c‐myc and EpCAM. Increased GATA5 expression by transfection with its expression vectors was also able to inhibit the cell growth, colony formation and capability of migration, invasion, while promoting apoptosis in HCC cells. Results revealed that GATA5 co‐localization with β‐catenin in the cytoplasm, preventing β‐catenin from entering the nucleus. Treatment with the specific Wnt/β‐catenin pathway inhibitor salinomycin was able to reduce the expression of β‐catenin and reprogramming genes. Salinomycin exerted a similar influence as GATA5, and siRNA‐GATA5 restored β‐catenin and reprogramming gene expression. This study demonstrates that an increase in the expression of GATA5 inhibits the expression of β‐catenin and reprogramming genes and suppresses tumour growth, colony formation, metastasis and invasion, while promoting apoptosis in HCC cells. The mechanism of GATA5 inhibiting the malignant behaviours of HCC cells may involve in the disruption of the Wnt/β‐catenin pathway and the reduction of reprogramming gene expression.

Vincosamide inhibits malignant behaviors of hepatocellular carcinoma cells by activating caspase-3 activity and blocking the PI3K/AKT signaling pathway

“…The cell viability ratio was calculated according to the following formula: (control group viable cells-treated groups viable cells)/control group viable cells×100%. The death of cells was evaluated by microscopy as previously described [14] Electron microscopy to observe mitochondria HLE, Bel 7402 and PLC/PRF/5 cells were cultured in RPMI 1640 medium supplemented with 10% FCS at 37 °C in a humidified atmosphere of 5% CO 2 . The cells were treated with Vinco (80 μg/ml) for 48 h and then harvested, washed with phosphate-buffered saline (PBS) two times (4 °C, 10 ml, 8 min), and transferred to 1.5-ml EP tubes.…”

“…HLE, Bel 7402 and PLC/PRF/5 cells were stained as described previously [14]. To observe the migration of caspase-3 molecules, cells were treated with Vinco (10 μg/ml and 80 μg/ml); to evaluate the expression and location of Src, Oct4 and PTEN, cells were treated with Vinco (80 μg/ml).…”

“…Nonimmune (IgG) was used as a negative control, and antigenic sites were identified using an SP9000 Polymer Detection System and a 3,3′-diaminobenzidine kit (ZSGB-BIO, Beijing, China). The standard protocols were performed in accordance with approved guidelines [14,17].…”

Vincosamide inhibits malignant behaviors of hepatocellular carcinoma cells by activating caspase-3 activity and blocking the PI3K/AKT signaling pathway

“…ve different visual elds was use to analyze the quantity of cell viability ratio, the cell viability ratio was calculated according to the following formula: (total cells-Trypan blue stained cells)/total cells × 100%. The procedure and calculations were based on as previously described [Feng et al, 2019].…”

“…Cell repair motility was evaluated using the following formula: cell repair ratio (%) =(distance at 0 h-distance at X h)/distance at 0 h × 100%, with X representing the time points of observation. The procedure was performed as previously described [Feng et al, 2019].…”