Loss of GATA5 expression due to gene promoter methylation induces growth and colony formation of hepatocellular carcinoma cells

Abstract

Abstract. GATA5 is a transcription factor that is capable of suppressing the development of various types of human cancer. The present study investigated the expression of GATA5 and GATA4, and examined their roles in the proliferation and colony formation ability of hepatocellular carcinoma (HCC) tissues and cells. The GATA4 and GATA5 expression levels and gene promoter methylation of HCC tissue samples from 38 patients and HCC cell lines were analyzed using reverse transcription-polymerase chain reaction (RT-PCR) and methylation-specific PCR (MSP), respectively. The effects of GATA4 and GATA5 overexpression on the proliferation and colony forming ability of HCC cells were also assessed using cell viability and colony formation assays. A luciferase reporter assay was utilized to investigate the transcriptional interaction of GATA4 and GATA5 with canonical Wnt signaling. The results indicated that the expression levels of GATA4 and GATA5 were lost or reduced following methylation of gene promoters in HCC tissues and cell lines. Treatment with a demethylating agent, 5-aza-2'-deoxycytidine (5-AZA), restored GATA4 and GATA5 expression in HCC cell lines. Furthermore, methylation of the GATA5 promoter was observed to be associated with the age of patients exhibiting HCC. Restoration of GATA4 and GATA5 expression inhibited colony formation and induced apoptosis of HCC cells in vitro. The present study concluded that the expression levels of GATA4 and GATA5 were reduced in HCC tissues and cell lines. Treatment with 5-AZA restored GATA4 and GATA5 expression in HCC cell lines, suppressing tumor cell growth and colony formation, as well as inducing apoptosis.

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

“…A previous study found that GATA4 and GATA5 might be involved in the regulation of the Wnt signalling pathway in HepG2 cells through Luciferase reporter assay. 5 In this study, we found that the expression of β-cat- Herein, we demonstrated for the first time that GATA5 was able to down-regulate the expression of β-catenin and the reprogramming genes p-Oct4, Nanog, Klf4, c-myc and EpCAM in HCC tissues and cells. To our surprise that GATA5 not only plays a role in inhibiting expression of β-catenin, but also blocking β-catenin enter the nucleus.…”

Disruption of the Gut Microbiota Attenuates Epithelial Ovarian Cancer Sensitivity to Cisplatin Therapy

“…SALL1 interacts with NANOG a well-established pluripotency transcription factor, to suppress differentiation (44). Conversely, SHOX2 and GATA5 genes that favor differentiation, were signi cantly decreased providing further evidence that ID8 tumor cells from ABX mice are more stem like and undifferentiated than ID8 tumor cells from H 2 O treated mice (41,46,55). Upon KEGG pathway analysis of enriched genes in the ABX group, multiple signaling pathways such as P13K/AKT/mTOR and WNT, were implicated and currently under investigation as targets for ovarian cancer therapies (49,56).…”

“…SHOX2 and GATA5 genes that promote cell differentiation were also signi cantly decreased (Fig. 5C) (39)(40)(41)(42)(43)(44)(45)(46). Upon KEGG pathway analysis of genes with a p-value < 0.01 in the ABX Placebo group vs. the H 2 O Placebo group, multiple signaling pathways were implicated including PI3-Akt, MAPK, WNT, and Hedgehog (Fig.…”

Methylation status of the promoter region of the human frizzled 9 gene in acute myeloid leukemia

“…PCR products were separated on 1.5% low-melting agarose gels, excised and then digested with β-agarase (New England Biolabs, Beverly, MA, USA). The digestion products were sub-cloned into the pMD18-T vector as previously described (31), and a minimum of 8 clones from each product were subjected to cycle sequencing (Applied Biosystems, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and analyzed using the ABI 310 sequencer (Applied Biosystems).…”