Reactivation of the homeotic tumor suppressor gene CDX2 by 5-aza-2′-deoxycytidine-induced demethylation inhibits cell proliferation and induces caspase-independent apoptosis in gastric cancer cells

Zhang, Jianfeng; Zhang, Jianguo; Kuai, Xiaoling; Zhang, Hong; Jiang, Wei; Ding, Weifeng; Li, Zeng-Li; Zhu, Huijun; Mao, Zhen-biao

doi:10.3892/etm.2013.901

Cited by 19 publications

(12 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Amplification was performed in a 7500 Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, USA) using the QuantiTect SYBR Green PCR kit (Qiagen, Hilden, Germany). PCR was performed as previously described using primers specific for β-actin and CDX2 (20). The β-actin gene was adopted as an internal control for all RT-qPCR reactions.…”

Section: Methodsmentioning

confidence: 99%

Nuclear transcription factor CDX2 inhibits gastric cancer-cell growth and reverses epithelial-to-mesenchymal transition in vitro and in vivo

Zhang

Qian

et al. 2015

Molecular Medicine Reports

Self Cite

View full text Add to dashboard Cite

The epithelial‑to‑mesenchymal transition (EMT) has been noted as a critical event in the early step of cancer metastasis. Recent studies showed that nuclear transcription factor caudal type homeobox transcription factor 2 (CDX2) is a prognostic factor, which acts as a marker of good outcome in gastric cancer (GC) patients. However, the association between CDX2 expression and EMT has remained to be fully elucidated. The present study reported that forced overexpression of CDX2 in MKN45/CDX2 cells inhibited GC‑cell growth and proliferation, and attenuated migration and invasion in vitro. Furthermore, MKN45/CDX2 cells exhibited a significant upregulation of E‑cadherin protein and a significant downregulation of vimentin protein expression. These results were further supported by in vivo tumorigenicity assays, which showed that CDX2 suppressed gastric tumor xenograft growth and inhibited EMT in nude mice. These results indicated that CDX2 is capable of inhibiting GC‑cell growth and invasion. CDX2 may participate in the process of EMT of GC cells by regulating the expression of the epithelial and mesenchymal proteins E‑cadherin and vimentin.

show abstract

Section: Methodsmentioning

confidence: 99%

Nuclear transcription factor CDX2 inhibits gastric cancer-cell growth and reverses epithelial-to-mesenchymal transition in vitro and in vivo

Zhang

Qian

et al. 2015

Molecular Medicine Reports

Self Cite

View full text Add to dashboard Cite

show abstract

“…Before exposure to Fe/Asc (200 µM/2 mM) for 6 h at 37°C, Caco-2/15 cells were pre-incubated with 0.25 mM 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), with ethanol as a vehicle, for 24 h. Four experimental cell groups were considered: control (with 0.25 mM ethanol as a vehicle), Fe/Asc (to induce OxS), Trolox (as antioxidant) and Trolox+Fe/Asc (to assess the direct contribution of OxS via its neutralisation by the antioxidant). For the studies of DNA methylation, Caco-2/15 cells were pre-incubated with 5-AZA at a concentration of 10 µM [28] for 54 h [28], [29], and 5 experimental cell groups were established: control (with ethanol as a vehicle), 5-AZA (to preclude methylation), Fe/Asc+5-AZA (to prevent methylation in the presence of OxS), Trolox+5-AZA (to evaluate the combined effect of antioxidant and demethylating agent), and Fe/Asc+Trolox+5-AZA (to evaluate the combined effect of antioxidant and demethylating agent in the presence of OxS).…”

Section: Methodsmentioning

confidence: 99%

Iron-Ascorbate-Mediated Lipid Peroxidation Causes Epigenetic Changes in the Antioxidant Defense in Intestinal Epithelial Cells: Impact on Inflammation

et al. 2013

View full text Add to dashboard Cite

IntroductionThe gastrointestinal tract is frequently exposed to noxious stimuli that may cause oxidative stress, inflammation and injury. Intraluminal pro-oxidants from ingested nutrients especially iron salts and ascorbic acid frequently consumed together, can lead to catalytic formation of oxygen-derived free radicals that ultimately overwhelm the cellular antioxidant defense and lead to cell damage.HypothesisSince the mechanisms remain sketchy, efforts have been exerted to evaluate the role of epigenetics in modulating components of endogenous enzymatic antioxidants in the intestine. To this end, Caco-2/15 cells were exposed to the iron-ascorbate oxygen radical-generating system.ResultsFe/Asc induced a significant increase in lipid peroxidation as reflected by the elevated formation of malondialdehyde along with the alteration of antioxidant defense as evidenced by raised superoxide dismutase 2 (SOD2) and diminished glutathione peroxidase (GPx) activities and genes. Consequently, there was an up-regulation of inflammatory processes illustrated by the activation of NF-κB transcription factor, the higher production of interleukin-6 and cycloxygenase-2 as well as the decrease of IκB. Assessment of promoter’s methylation revealed decreased levels for SOD2 and increased degree for GPx2. On the other hand, pre-incubation of Caco-2/15 cells with 5-Aza-2′-deoxycytidine, a demethylating agent, or Trolox antioxidant normalized the activities of SOD2 and GPx, reduced lipid peroxidation and prevented inflammation.ConclusionRedox and inflammatory modifications in response to Fe/Asc -mediated lipid peroxidation may implicate epigenetic methylation.

show abstract

“…The DNA methyltransferase (DNMT) inhibitor, 5-Aza-2'-deoxycytidine (5-Aza-CdR) is able to restore the expression of genes that induce growth arrest and apoptosis in tumor cells (12,13). In the F6 tumor cell line, the effect of 5-Aza-CdR on the cell cycle and cell apoptosis requires additional investigation.…”

Section: Introductionmentioning

confidence: 99%

Methylation status of the FHIT gene in the transformed human mesenchymal F6 stem cell line

Gao

Wang

et al. 2015

Oncology Letters

View full text Add to dashboard Cite

Abstract. The fragile histidine triad (FHIT) gene is known to be a tumor suppressor gene and the abnormal methylation of FHIT has been identified in leukemia and several solid tumors. The transformation of the tumor F6 cell line from human fetal mesenchymal stem cells (FMSCs) was first reported in a previous study that also identified the presence of a population of cancer stem cells in the F6 cell line. However, the existence of the epigenetic changes during the transformation process have yet to be elucidated. To confirm the role of the FHIT gene in the transformation process of FMSC, the expression level and methylation status of the FHIT gene was examined in F6 tumor cells and FMSCs. Additionally, the alteration in cell morphology, the cell cycle and apoptosis in F6 cells following 5-Aza-CdR treatment was assessed. It was found that the FHIT gene was expressed in FMSCs, but not in F6 cells. The methylation-specific PCR results demonstrated that the promoter methylation of FHIT genes existed in the F6 cell line. Subsequent to treatment with 5-Aza-CdR the expression of FHIT genes was restored in F6 cells. In addition, the morphology of F6 cells was altered, and the cell cycle was arrested in the G 2 phase, with the initiation of apoptosis.Overall, the present findings demonstrated that the FHIT gene was methylated in F6 cells and demethylation treatment lead to changes in the biological characteristics, thereby promoting the apoptosis of F6 cells. FHIT gene methylation may be one of the molecular events involved in the development and transformation of FMSCs into F6 tumor cells. IntroductionMesenchymal stem cells (MSCs) are pluripotent adult stromal cells that possess the ability to differentiate into bone, cartilage and adipose tissues, and the cells exhibit promising potential for regenerative medicine (1). An increasing number of studies have demonstrated that MSCs demonstrate potential therapeutic effects for multiple diseases, including liver injury, ischemia/reperfusion-induced acute kidney injury and acute myocardial infarction (2-4). However, transplantation therapy using MSCs continues to be subject to investigation, particularly into the safety of MSCs in clinical application. In a previous study that aimed to determine the induction of MSC differentiation and immune function, tumorigenesis was observed unexpectedly and a cloned a novel tumor cell line, the F6 cell line, transformed from human fetal bone marrow MSCs (5). These findings initiated interest into investigating the mechanism of mutation in fetal MSCs (FMSCs).Previous studies have demonstrated that epigenetic and genetic alterations markedly contribute to tumorigenesis. The hypermethylation of tumor suppressor genes has been detected in various types of tumors (6). Ohta et al first reported the tumor suppressor gene fragile histidine triad (FHIT), providing the molecular evidence for the association between fragile sites and cancer (7). The loss or reduction of FHIT expression has been revealed to be important in the initiation of tumorige...

show abstract

Reactivation of the homeotic tumor suppressor gene CDX2 by 5-aza-2′-deoxycytidine-induced demethylation inhibits cell proliferation and induces caspase-independent apoptosis in gastric cancer cells

Cited by 19 publications

References 44 publications

Nuclear transcription factor CDX2 inhibits gastric cancer-cell growth and reverses epithelial-to-mesenchymal transition in vitro and in vivo

Nuclear transcription factor CDX2 inhibits gastric cancer-cell growth and reverses epithelial-to-mesenchymal transition in vitro and in vivo

Iron-Ascorbate-Mediated Lipid Peroxidation Causes Epigenetic Changes in the Antioxidant Defense in Intestinal Epithelial Cells: Impact on Inflammation

Methylation status of the FHIT gene in the transformed human mesenchymal F6 stem cell line

Contact Info

Product

Resources

About