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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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Functional enrichment of the differentially expressed genes, identified between SGH-44/ADM and SGH-44 cells (P<0.05). Sig GO, significant gene ontology; SHG-44/ADM, human glioma cell line SHG-44 with adriamycin resistance.
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f4-mmr-11-01-0043: Functional enrichment of the differentially expressed genes, identified between SGH-44/ADM and SGH-44 cells (P<0.05). Sig GO, significant gene ontology; SHG-44/ADM, human glioma cell line SHG-44 with adriamycin resistance.

Mentions: A functional enrichment analysis was performed using the Web-based Gene Set Analysis Toolkit (33,34) and 74 significantly enhanced functions were identified (Fig. 4; FDR < 0.05). Immune-associated reactions, including the adaptive immune response based on somatic recombination of immune receptors built from immunoglobulin superfamily domains, lymphocyte chemotaxis across the high endothelial venule and positive regulation of cytotoxic T-cell differentiation were significantly enriched (enrichment = 131; P<0.05).


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

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

Functional enrichment of the differentially expressed genes, identified between SGH-44/ADM and SGH-44 cells (P<0.05). Sig GO, significant gene ontology; SHG-44/ADM, human glioma cell line SHG-44 with adriamycin resistance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4-mmr-11-01-0043: Functional enrichment of the differentially expressed genes, identified between SGH-44/ADM and SGH-44 cells (P<0.05). Sig GO, significant gene ontology; SHG-44/ADM, human glioma cell line SHG-44 with adriamycin resistance.
Mentions: A functional enrichment analysis was performed using the Web-based Gene Set Analysis Toolkit (33,34) and 74 significantly enhanced functions were identified (Fig. 4; FDR < 0.05). Immune-associated reactions, including the adaptive immune response based on somatic recombination of immune receptors built from immunoglobulin superfamily domains, lymphocyte chemotaxis across the high endothelial venule and positive regulation of cytotoxic T-cell differentiation were significantly enriched (enrichment = 131; P<0.05).

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