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Association between DNA methylation and multidrug resistance in human glioma SHG-44 cells.

Chen J, Xu ZY, Wang F - Mol Med Rep (2014)

Bottom Line: SGH-44/ADM cells showed little difference as compared with parental cells, except that SGH-44/ADM cells were bigger in size with a wizened nucleus.The expression of COX-2, PKCα and P-glycoprotein (Pgp) was not found to be associated with DNA methylation.Genes including SNAP47, VAMP4 and VAMP3 may serve as the downstream effectors of Pgp, COX-2 or PKCα; however, further experiments are required to verify these observations.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China.

ABSTRACT
The aim of the present study was to evaluate the association between DNA methylation and multidrug resistance (MDR) in glioma and identify novel effectors responsible for MDR in human gliomas. An MDR glioma cell line, SGH-44/ADM, was developed using adriamycin (ADM) impulse treatment. Cryopreservation, recovery and withdrawal were performed to evaluate the stability of SGH-44/ADM cells. The adherence rate and cellular morphology were observed by microscopy, and the cell growth curve and doubling time were determined. DNA methylation was analyzed using a methylated DNA immunoprecipitation microarray chip (MeDIP-Chip). The cell cycle, Rh123 ingestion and exudation, and SGH-44/ADM apoptosis were analyzed by flow cytometry. SGH-44/ADM cells showed little difference as compared with parental cells, except that SGH-44/ADM cells were bigger in size with a wizened nucleus. Compared to SGH-44 cells, a larger proportion of SGH-44/ADM cells remained in G1 and S phase, as measured by flow cytometry. The MDR of SGH-44/ADM was associated with the upregulation of multi-drug resistance 1, prostaglandin-endoperoxide synthase 2 (COX-2); protein kinase C α (PKCα); however, the expression of these genes was not associated with DNA methylation. In the MeDIP-Chip analysis, 74 functions were markedly enhanced, and seven significant pathways were observed. Genes including SNAP47, ARRB2, PARD6B, TGFB1, VPS4B and CBLB were identified by gene ontology analysis. The predominant molecular mechanism of MDR in SGH-44/ADM cells was identified as exocytosis and efflux. The expression of COX-2, PKCα and P-glycoprotein (Pgp) was not found to be associated with DNA methylation. Genes including SNAP47, VAMP4 and VAMP3 may serve as the downstream effectors of Pgp, COX-2 or PKCα; however, further experiments are required to verify these observations.

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Pathway enrichment in MeDIP-Chip.
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f5-mmr-11-01-0043: Pathway enrichment in MeDIP-Chip.

Mentions: Pathway enrichment analysis of all genes involved in MeDIP-Chip was performed using the KEGG automatic annotation server. A total of seven significant pathways with P<0.05 were enriched (Fig. 5). The most significant pathway was synaptosomal-associated protein (SNAP) receptor (SNARE) interaction in vesicular transport (path_id=4130), where P=0.005. Three genes were involved in this pathway, including SNAP47, vesicle associated membrane protein (VAMP)4 and VAMP3. The other significantly enriched pathways included sphingolipid metabolism, endocytosis, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, collecting duct acid secretion, pentose phosphate pathway and glycine, serine and threonine metabolism. Genes associated with endocytosis included charged multivesicular body protein 1b (CHMP1B), arrestin β2 (ARRB2), par-6 family cell polarity regulator β (PARD6B), transforming growth factor β1 (TGFB1), vacuolar protein sorting 4 homolog B (VPS4B) and cbl proto-oncogene, E3 ubiquitin protein ligase B (CBLB).


Association between DNA methylation and multidrug resistance in human glioma SHG-44 cells.

Chen J, Xu ZY, Wang F - Mol Med Rep (2014)

Pathway enrichment in MeDIP-Chip.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4237088&req=5

f5-mmr-11-01-0043: Pathway enrichment in MeDIP-Chip.
Mentions: Pathway enrichment analysis of all genes involved in MeDIP-Chip was performed using the KEGG automatic annotation server. A total of seven significant pathways with P<0.05 were enriched (Fig. 5). The most significant pathway was synaptosomal-associated protein (SNAP) receptor (SNARE) interaction in vesicular transport (path_id=4130), where P=0.005. Three genes were involved in this pathway, including SNAP47, vesicle associated membrane protein (VAMP)4 and VAMP3. The other significantly enriched pathways included sphingolipid metabolism, endocytosis, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, collecting duct acid secretion, pentose phosphate pathway and glycine, serine and threonine metabolism. Genes associated with endocytosis included charged multivesicular body protein 1b (CHMP1B), arrestin β2 (ARRB2), par-6 family cell polarity regulator β (PARD6B), transforming growth factor β1 (TGFB1), vacuolar protein sorting 4 homolog B (VPS4B) and cbl proto-oncogene, E3 ubiquitin protein ligase B (CBLB).

Bottom Line: SGH-44/ADM cells showed little difference as compared with parental cells, except that SGH-44/ADM cells were bigger in size with a wizened nucleus.The expression of COX-2, PKCα and P-glycoprotein (Pgp) was not found to be associated with DNA methylation.Genes including SNAP47, VAMP4 and VAMP3 may serve as the downstream effectors of Pgp, COX-2 or PKCα; however, further experiments are required to verify these observations.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China.

ABSTRACT
The aim of the present study was to evaluate the association between DNA methylation and multidrug resistance (MDR) in glioma and identify novel effectors responsible for MDR in human gliomas. An MDR glioma cell line, SGH-44/ADM, was developed using adriamycin (ADM) impulse treatment. Cryopreservation, recovery and withdrawal were performed to evaluate the stability of SGH-44/ADM cells. The adherence rate and cellular morphology were observed by microscopy, and the cell growth curve and doubling time were determined. DNA methylation was analyzed using a methylated DNA immunoprecipitation microarray chip (MeDIP-Chip). The cell cycle, Rh123 ingestion and exudation, and SGH-44/ADM apoptosis were analyzed by flow cytometry. SGH-44/ADM cells showed little difference as compared with parental cells, except that SGH-44/ADM cells were bigger in size with a wizened nucleus. Compared to SGH-44 cells, a larger proportion of SGH-44/ADM cells remained in G1 and S phase, as measured by flow cytometry. The MDR of SGH-44/ADM was associated with the upregulation of multi-drug resistance 1, prostaglandin-endoperoxide synthase 2 (COX-2); protein kinase C α (PKCα); however, the expression of these genes was not associated with DNA methylation. In the MeDIP-Chip analysis, 74 functions were markedly enhanced, and seven significant pathways were observed. Genes including SNAP47, ARRB2, PARD6B, TGFB1, VPS4B and CBLB were identified by gene ontology analysis. The predominant molecular mechanism of MDR in SGH-44/ADM cells was identified as exocytosis and efflux. The expression of COX-2, PKCα and P-glycoprotein (Pgp) was not found to be associated with DNA methylation. Genes including SNAP47, VAMP4 and VAMP3 may serve as the downstream effectors of Pgp, COX-2 or PKCα; however, further experiments are required to verify these observations.

Show MeSH
Related in: MedlinePlus