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Identification of proteins responsible for adriamycin resistance in breast cancer cells using proteomics analysis.

Wang Z, Liang S, Lian X, Liu L, Zhao S, Xuan Q, Guo L, Liu H, Yang Y, Dong T, Liu Y, Liu Z, Zhang Q - Sci Rep (2015)

Bottom Line: Basing on the human protein-protein interactions (PPI), we have retrieved the associated functional interaction networks for the DEPs and analyzed the biological functions.The identified proteins in biological networks served to resistant drug and to select critical candidates for validation analyses by western blot.The glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamyl cyclotransferase (GGCT), isocitrate dehydrogenase 1 (NADP+,soluble)(IDH1), isocitrate dehydrogenase 2 (NADP+,mitochondrial) (IDH2) and glutathione S-transferase pi 1(GSTP1), five of the critical components of GSH pathway, contribute to chemoresistance.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Medical Oncology, The Third Affiliated Hospital of Harbin Medical University, Haping Road 150 of Nangang District, Harbin 150081, Heilongjiang Province, China [2] Department of Medical Oncology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China.

ABSTRACT
Chemoresistance is a poor prognostic factor in breast cancer and is a major obstacle to the successful treatment of patients receiving chemotherapy. However, the precise mechanism of resistance remains unclear. In this study, a pair of breast cancer cell lines, MCF-7 and its adriamycin-resistant counterpart MCF-7/ADR was used to examine resistance-dependent cellular responses and to identify potential therapeutic targets. We applied nanoflow liquid chromatography (nLC) and tandem mass tags (TmT) quantitative mass spectrometry to distinguish the differentially expressed proteins (DEPs) between the two cell lines. Bioinformatics analyses were used to identify functionally active proteins and networks. 80 DEPs were identified with either up- or down-regulation. Basing on the human protein-protein interactions (PPI), we have retrieved the associated functional interaction networks for the DEPs and analyzed the biological functions. Six different signaling pathways and most of the DEPs strongly linked to chemoresistance, invasion, metastasis development, proliferation, and apoptosis. The identified proteins in biological networks served to resistant drug and to select critical candidates for validation analyses by western blot. The glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamyl cyclotransferase (GGCT), isocitrate dehydrogenase 1 (NADP+,soluble)(IDH1), isocitrate dehydrogenase 2 (NADP+,mitochondrial) (IDH2) and glutathione S-transferase pi 1(GSTP1), five of the critical components of GSH pathway, contribute to chemoresistance.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis confirmed changes in protein expression initially identified by quantitative proteomics method.The expression of DEPs was focused on GSH metabolism pathway. G6PD, GGCT, IDH1 and IDH2 were found down–regulated in MCF-7/ADR cells compared with MCF-7 cells. Up-regulated expression was observed for GSTP1. GAPDH was used as the loading control.
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f5: Western blot analysis confirmed changes in protein expression initially identified by quantitative proteomics method.The expression of DEPs was focused on GSH metabolism pathway. G6PD, GGCT, IDH1 and IDH2 were found down–regulated in MCF-7/ADR cells compared with MCF-7 cells. Up-regulated expression was observed for GSTP1. GAPDH was used as the loading control.

Mentions: The expression of G6PD, GGCT, IDH1, IDH2 and GSTP1 were further validated by western blot. Consistent with the observations in proteomics analysis, G6PD, GGCT, IDH1 and IDH2 were found down-regulated in MCF-7/ADR cells compared with MCF-7 cells, and GSTP1 was found up-regulated in MCF-7/ADR cells compared with MCF-7 cells (Fig. 5).


Identification of proteins responsible for adriamycin resistance in breast cancer cells using proteomics analysis.

Wang Z, Liang S, Lian X, Liu L, Zhao S, Xuan Q, Guo L, Liu H, Yang Y, Dong T, Liu Y, Liu Z, Zhang Q - Sci Rep (2015)

Western blot analysis confirmed changes in protein expression initially identified by quantitative proteomics method.The expression of DEPs was focused on GSH metabolism pathway. G6PD, GGCT, IDH1 and IDH2 were found down–regulated in MCF-7/ADR cells compared with MCF-7 cells. Up-regulated expression was observed for GSTP1. GAPDH was used as the loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4377623&req=5

f5: Western blot analysis confirmed changes in protein expression initially identified by quantitative proteomics method.The expression of DEPs was focused on GSH metabolism pathway. G6PD, GGCT, IDH1 and IDH2 were found down–regulated in MCF-7/ADR cells compared with MCF-7 cells. Up-regulated expression was observed for GSTP1. GAPDH was used as the loading control.
Mentions: The expression of G6PD, GGCT, IDH1, IDH2 and GSTP1 were further validated by western blot. Consistent with the observations in proteomics analysis, G6PD, GGCT, IDH1 and IDH2 were found down-regulated in MCF-7/ADR cells compared with MCF-7 cells, and GSTP1 was found up-regulated in MCF-7/ADR cells compared with MCF-7 cells (Fig. 5).

Bottom Line: Basing on the human protein-protein interactions (PPI), we have retrieved the associated functional interaction networks for the DEPs and analyzed the biological functions.The identified proteins in biological networks served to resistant drug and to select critical candidates for validation analyses by western blot.The glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamyl cyclotransferase (GGCT), isocitrate dehydrogenase 1 (NADP+,soluble)(IDH1), isocitrate dehydrogenase 2 (NADP+,mitochondrial) (IDH2) and glutathione S-transferase pi 1(GSTP1), five of the critical components of GSH pathway, contribute to chemoresistance.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Medical Oncology, The Third Affiliated Hospital of Harbin Medical University, Haping Road 150 of Nangang District, Harbin 150081, Heilongjiang Province, China [2] Department of Medical Oncology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China.

ABSTRACT
Chemoresistance is a poor prognostic factor in breast cancer and is a major obstacle to the successful treatment of patients receiving chemotherapy. However, the precise mechanism of resistance remains unclear. In this study, a pair of breast cancer cell lines, MCF-7 and its adriamycin-resistant counterpart MCF-7/ADR was used to examine resistance-dependent cellular responses and to identify potential therapeutic targets. We applied nanoflow liquid chromatography (nLC) and tandem mass tags (TmT) quantitative mass spectrometry to distinguish the differentially expressed proteins (DEPs) between the two cell lines. Bioinformatics analyses were used to identify functionally active proteins and networks. 80 DEPs were identified with either up- or down-regulation. Basing on the human protein-protein interactions (PPI), we have retrieved the associated functional interaction networks for the DEPs and analyzed the biological functions. Six different signaling pathways and most of the DEPs strongly linked to chemoresistance, invasion, metastasis development, proliferation, and apoptosis. The identified proteins in biological networks served to resistant drug and to select critical candidates for validation analyses by western blot. The glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamyl cyclotransferase (GGCT), isocitrate dehydrogenase 1 (NADP+,soluble)(IDH1), isocitrate dehydrogenase 2 (NADP+,mitochondrial) (IDH2) and glutathione S-transferase pi 1(GSTP1), five of the critical components of GSH pathway, contribute to chemoresistance.

No MeSH data available.


Related in: MedlinePlus