Limits...
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

The expression of identified differentially expressed molecules in endometrial carcinoma (tumor) and pericarcinous tissue (normal).Notes: The mRNA expression of (A) CCT7, (B) HSPA8, (C) PCBP2, (D) LONP1, (E) PFN1, and (F) EEF2 was detected by qRT-PCR (n=10). (G) The expression of HSPA8 in protein level was also detected by Western blot (n=4). *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs normal.Abbreviation: qRT-PCR, quantitative real-time polymerase chain reaction.
© Copyright Policy
Related In: Results  -  Collection

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

f3-ott-9-2169: The expression of identified differentially expressed molecules in endometrial carcinoma (tumor) and pericarcinous tissue (normal).Notes: The mRNA expression of (A) CCT7, (B) HSPA8, (C) PCBP2, (D) LONP1, (E) PFN1, and (F) EEF2 was detected by qRT-PCR (n=10). (G) The expression of HSPA8 in protein level was also detected by Western blot (n=4). *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs normal.Abbreviation: qRT-PCR, quantitative real-time polymerase chain reaction.

Mentions: From the proteomic data with iTRAQ and LC-MS/MS, we identified a list of proteins. We further validated them by qRT-PCR. Our results revealed that the mRNA expression of CCT7 (Figure 3A), HSPA8 (Figure 3B), PCBP2 (Figure 3C), LONP1 (Figure 3D), PFN1 (Figure 3E), and EEF2 (Figure 3F) was significantly higher in endometrial carcinoma tissues than in the normal pericarcinous tissue (n=10). The upregulation of these molecules in endometrial carcinoma was in concordance with the iTRAQ results. We found that HSPA8 was most significantly upregulated in mRNA levels in endometrial carcinoma tissues (P<0.0001). So, the HSPA8 was selected for further investigation. By Western blot, we also observe the upregulation of HSPA8 in the endometrial carcinoma tissues (Figure 3G).


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)

The expression of identified differentially expressed molecules in endometrial carcinoma (tumor) and pericarcinous tissue (normal).Notes: The mRNA expression of (A) CCT7, (B) HSPA8, (C) PCBP2, (D) LONP1, (E) PFN1, and (F) EEF2 was detected by qRT-PCR (n=10). (G) The expression of HSPA8 in protein level was also detected by Western blot (n=4). *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs normal.Abbreviation: qRT-PCR, quantitative real-time polymerase chain reaction.
© Copyright Policy
Related In: Results  -  Collection

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

f3-ott-9-2169: The expression of identified differentially expressed molecules in endometrial carcinoma (tumor) and pericarcinous tissue (normal).Notes: The mRNA expression of (A) CCT7, (B) HSPA8, (C) PCBP2, (D) LONP1, (E) PFN1, and (F) EEF2 was detected by qRT-PCR (n=10). (G) The expression of HSPA8 in protein level was also detected by Western blot (n=4). *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs normal.Abbreviation: qRT-PCR, quantitative real-time polymerase chain reaction.
Mentions: From the proteomic data with iTRAQ and LC-MS/MS, we identified a list of proteins. We further validated them by qRT-PCR. Our results revealed that the mRNA expression of CCT7 (Figure 3A), HSPA8 (Figure 3B), PCBP2 (Figure 3C), LONP1 (Figure 3D), PFN1 (Figure 3E), and EEF2 (Figure 3F) was significantly higher in endometrial carcinoma tissues than in the normal pericarcinous tissue (n=10). The upregulation of these molecules in endometrial carcinoma was in concordance with the iTRAQ results. We found that HSPA8 was most significantly upregulated in mRNA levels in endometrial carcinoma tissues (P<0.0001). So, the HSPA8 was selected for further investigation. By Western blot, we also observe the upregulation of HSPA8 in the endometrial carcinoma tissues (Figure 3G).

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