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Identification of HSPA8 as a candidate biomarker for endometrial carcinoma by using iTRAQ-based proteomic analysis.

Shan N, Zhou W, Zhang S, Zhang Y - Onco Targets Ther (2016)

Bottom Line: Totally, we screened 1,266 proteins.We further validated the overexpression of HSPA8 by using immunoblot analysis.The depletion of HSPA8 siRNAs significantly reduced cell proliferation, promoted cell apoptosis, and suppressed cell growth in both cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetric and Gynecology, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT
Although there are advances in diagnostic, predictive, and therapeutic strategies, discovering protein biomarker for early detection is required for improving the survival rate of the patients with endometrial carcinoma. In this study, we identify proteins that are differentially expressed between the Stage I endometrial carcinoma and the normal pericarcinous tissues by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. Totally, we screened 1,266 proteins. Among them, 103 proteins were significantly overexpressed, and 30 were significantly downexpressed in endometrial carcinoma. Using the bioinformatics analysis, we identified a list of proteins that might be closely associated with endometrial carcinoma, including CCT7, HSPA8, PCBP2, LONP1, PFN1, and EEF2. We validated the gene overexpression of these molecules in the endometrial carcinoma tissues and found that HSPA8 was most significantly upregulated. We further validated the overexpression of HSPA8 by using immunoblot analysis. Then, HSPA8 siRNA was transferred into the endometrial cancer cells RL-95-2 and HEC-1B. The depletion of HSPA8 siRNAs significantly reduced cell proliferation, promoted cell apoptosis, and suppressed cell growth in both cell lines. Taken together, HSPA8 plays a vital role in the development of endometrial carcinoma. HSPA8 is a candidate biomarker for early diagnosis and therapy of Stage I endometrial carcinoma.

No MeSH data available.


Related in: MedlinePlus

Representative figure showing differential isotopic labeling and LC-MS/MS.Notes: Representative data for an upregulated protein, HSPA8. The continuous series of the b- and y-ions used for the identification of the peptide fragment.Abbreviation: LC-MS/MS, Liquid chromatography tandom-mass spectrometry.
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f2-ott-9-2169: Representative figure showing differential isotopic labeling and LC-MS/MS.Notes: Representative data for an upregulated protein, HSPA8. The continuous series of the b- and y-ions used for the identification of the peptide fragment.Abbreviation: LC-MS/MS, Liquid chromatography tandom-mass spectrometry.

Mentions: The differentially expressed proteins in endometrial carcinoma were further assigned to 14 KEGG pathways (P<0.05; Table 1), which include pathways for legionellosis, endocytosis, pathogenic Escherichia coli infection, shigellosis, glycolysis/gluconeogenesis, cardiac muscle contraction, fructose and mannose metabolism, African trypanosomiasis, bacterial invasion of epithelial cells, hyper-trophic cardiomyopathy, viral carcinogenesis, phagosome, protein processing in endoplasmic reticulum, and amino sugar and nucleotide sugar metabolism. Among them, selected molecules associated with endometrial carcinoma were further identified by LC-MS/MS spectra, including CCT7, heat shock 70 kDa protein 8 (HSPA8), PCBP2, LONP1, PFN1 and EEF2, which are listed in Table 2. The representative LC-MS/MS spectra of the molecule HSPA8 has been shown in Figure 2.


Identification of HSPA8 as a candidate biomarker for endometrial carcinoma by using iTRAQ-based proteomic analysis.

Shan N, Zhou W, Zhang S, Zhang Y - Onco Targets Ther (2016)

Representative figure showing differential isotopic labeling and LC-MS/MS.Notes: Representative data for an upregulated protein, HSPA8. The continuous series of the b- and y-ions used for the identification of the peptide fragment.Abbreviation: LC-MS/MS, Liquid chromatography tandom-mass spectrometry.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835145&req=5

f2-ott-9-2169: Representative figure showing differential isotopic labeling and LC-MS/MS.Notes: Representative data for an upregulated protein, HSPA8. The continuous series of the b- and y-ions used for the identification of the peptide fragment.Abbreviation: LC-MS/MS, Liquid chromatography tandom-mass spectrometry.
Mentions: The differentially expressed proteins in endometrial carcinoma were further assigned to 14 KEGG pathways (P<0.05; Table 1), which include pathways for legionellosis, endocytosis, pathogenic Escherichia coli infection, shigellosis, glycolysis/gluconeogenesis, cardiac muscle contraction, fructose and mannose metabolism, African trypanosomiasis, bacterial invasion of epithelial cells, hyper-trophic cardiomyopathy, viral carcinogenesis, phagosome, protein processing in endoplasmic reticulum, and amino sugar and nucleotide sugar metabolism. Among them, selected molecules associated with endometrial carcinoma were further identified by LC-MS/MS spectra, including CCT7, heat shock 70 kDa protein 8 (HSPA8), PCBP2, LONP1, PFN1 and EEF2, which are listed in Table 2. The representative LC-MS/MS spectra of the molecule HSPA8 has been shown in Figure 2.

Bottom Line: Totally, we screened 1,266 proteins.We further validated the overexpression of HSPA8 by using immunoblot analysis.The depletion of HSPA8 siRNAs significantly reduced cell proliferation, promoted cell apoptosis, and suppressed cell growth in both cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetric and Gynecology, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT
Although there are advances in diagnostic, predictive, and therapeutic strategies, discovering protein biomarker for early detection is required for improving the survival rate of the patients with endometrial carcinoma. In this study, we identify proteins that are differentially expressed between the Stage I endometrial carcinoma and the normal pericarcinous tissues by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. Totally, we screened 1,266 proteins. Among them, 103 proteins were significantly overexpressed, and 30 were significantly downexpressed in endometrial carcinoma. Using the bioinformatics analysis, we identified a list of proteins that might be closely associated with endometrial carcinoma, including CCT7, HSPA8, PCBP2, LONP1, PFN1, and EEF2. We validated the gene overexpression of these molecules in the endometrial carcinoma tissues and found that HSPA8 was most significantly upregulated. We further validated the overexpression of HSPA8 by using immunoblot analysis. Then, HSPA8 siRNA was transferred into the endometrial cancer cells RL-95-2 and HEC-1B. The depletion of HSPA8 siRNAs significantly reduced cell proliferation, promoted cell apoptosis, and suppressed cell growth in both cell lines. Taken together, HSPA8 plays a vital role in the development of endometrial carcinoma. HSPA8 is a candidate biomarker for early diagnosis and therapy of Stage I endometrial carcinoma.

No MeSH data available.


Related in: MedlinePlus