Effect of methylation inhibitor on EphB4 gene expression, proliferation and apoptosis in CEM cells / 中国当代儿科杂志
Chinese Journal of Contemporary Pediatrics
; (12): 205-209, 2012.
Article
en Zh
| WPRIM
| ID: wpr-320684
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WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study the regulation of methylation inhibitor 5-aza-2'-deoxycytidine on transcription of EphB4 gene and effects on the proliferation and apoptosis of human acute lymphocyte leukemia cell line CEM.</p><p><b>METHODS</b>Bisulfite sequencing PCR was used to detect CpG island methylation density in EphB4 promoter. The expression of EphB4 mRNA and protein was determined by Q-PCR and Western blot. MTS assay and flow cytometry were used to detect the apoptosis of CEM cells after treatment with different concentrations of 5-aza-2'-deoxycytidine (1.0, 2.5 and 5 μmol/L).</p><p><b>RESULTS</b>Methylation of EphB4 gene promoter was detected in CEM cells (31.4%). The methylation level of EphB4 gene was down-regulated after treatment with various concentrations of 5-aza-2'-deoxycytidine. The EphB4 mRNA and protein expression in CEM cells increased after 5-aza-2'-deoxycytidine treatment. 5-Aza-2'-deoxycytidine significantly inhibited the cell growth in dose and time dependent manners. Early apoptosis rates of CEM cells increased from 4.1% to 24.8% 96 hrs after 5-aza-2'-deoxycytidine treatment. CEM cells in G1 phase decreased from 62.4% to 46.8%, cells in G2 phase increased from 2.1% to 16.2%, and CEM cells were arrested in G2 phase after treatment with 5 μmol/L 5-aza-2'-deoxycytidine for 96 hrs.</p><p><b>CONCLUSIONS</b>5-Aza-2'-deoxycytidine, an inhibitor of specific methylation transferase, can induce expression of the silent EphB4 gene in CEM cells, inhibit the proliferation of leukemia cells and induce cell apoptosis.</p>
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Índice:
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Asunto principal:
Patología
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Farmacología
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Azacitidina
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ARN Mensajero
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Metilasas de Modificación del ADN
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Ciclo Celular
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Apoptosis
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Metilación de ADN
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Receptor EphB4
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Línea Celular Tumoral
Límite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Contemporary Pediatrics
Año:
2012
Tipo del documento:
Article