Dataset Information

Identification of C6-ceramide-interacting proteins in D6P2T Schwannoma cells.

ABSTRACT: In this study, we have utilized affinity purification with a synthetic biotin-ceramide and LC-MS/MS to identify potential ceramide-interacting proteins in D6P2T Schwannoma cells.

INSTRUMENT(S): instrument model, LTQ

ORGANISM(S): Rattus Rattus (black Rat)

TISSUE(S): Rt4-d6p2t Cell

SUBMITTER: Venkatesh Kota

PROVIDER: PRD000566 | Pride | 2015-09-07

REPOSITORIES: Pride

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			Action	DRS
	PRIDE_Exp_Complete_Ac_19308.pride.mgf.gz	Mgf
	PRIDE_Exp_Complete_Ac_19308.pride.mztab.gz	Mztab
	PRIDE_Exp_Complete_Ac_19308.xml.gz	Xml
	PRIDE_Exp_Complete_Ac_19309.pride.mgf.gz	Mgf
	PRIDE_Exp_Complete_Ac_19309.pride.mztab.gz	Mztab

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Publications

Identification of C(6) -ceramide-interacting proteins in D6P2T Schwannoma cells.

Kota Venkatesh V Szulc Zdzislaw M ZM Hama Hiroko H

Proteomics 20120701 13

Ceramide is a bioactive molecule involved in numerous cell signaling pathways that are associated with cell cycle control, differentiation, senescence, and apoptosis. Although substantial knowledge about ceramide-regulated pathways has accumulated in the past decade, molecular mechanisms of ceramide action remain poorly understood, primarily due to limited information about ceramide-binding proteins. In the present study, we used affinity purification with a synthetic biotin-conjugated C(6) -cer ...[more]

PMID: 22623228

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Project description:We have used human umbilical vein endothelial cell line (HUVEC) as a model to investigate the effect of ultrasound (US) activated micro-bubbles on sensitizing the tumour response to radiation and to check the implication of this approach on gene expression. The use of micro-bubbles in cancer therapy is a novel approach that is being recently investigated, and patterns of gene expressions were not yet characterized. Gene analysis has identified about 19,264 genes; where, 239 – 517 genes have shown more than two folds up-regulation in response to the different treatments. This included a number of genes that are known to be involved in apoptosis and ceramide-induced apoptosis pathways, such as Sphingomyelin phosphodiesterase 2, neutral sphingomyelinase (SMPD2), sphingomyelin phosphodiesterase 1, acid lysosomal (ASM; SMPD1), UDP-galactose ceramide galactosyltransferase (UGT8), cytochrome c oxidase (COX6B1), Caspase-9 and mitogen-activated protein kinase kinase 1 (MAP2K1). To verify these results, we have performed Real time RT-PCR, which indicated an up-regulation that ranged from 2 to 200 folds, specifically in the combined treatment of US, 3.3% micro-bubbles and 8 Gy. The observed higher levels of expression of UGT8, SMPD2 and Caspase 9-alpha when compared to the control or to radiation only indicated the involvement of ceramide, which possibly induces apoptosis . These results support the hypothesis that micro-bubbles play a role in increasing sensitization to radiation, and can prove to be an effective cancer therapeutic approach. Gene analysis has identified about 19,264 genes; where, 239 – 517 genes have shown more than two folds up-regulation in response to the different treatments. This included a number of genes that are known to be involved in apoptosis and ceramide-induced apoptosis pathways, such as Sphingomyelin phosphodiesterase 2, neutral sphingomyelinase (SMPD2), sphingomyelin phosphodiesterase 1, acid lysosomal (ASM; SMPD1), UDP-galactose ceramide galactosyltransferase (UGT8), cytochrome c oxidase (COX6B1), Caspase-9 and mitogen-activated protein kinase kinase 1 (MAP2K1). To verify these results, we have performed Real time RT-PCR, which indicated an up-regulation that ranged from 2 to 200 folds, specifically in the combined treatment of US, 3.3% micro-bubbles and 8 Gy. The observed higher levels of expression of UGT8, SMPD2 and Caspase 9-alpha when compared to the control or to radiation only indicated the involvement of ceramide, which possibly induces apoptosis . These results support the hypothesis that micro-bubbles play a role in increasing sensitization to radiation, and can prove to be an effective cancer therapeutic approach.