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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

Screening of HSPA8 siRNA for knockdown of HSPA8 in RL-95-2 and HEC-1B cells.Notes: RL-95-2 cells were transfected with 50 µM, 100 µM, and 200 µM HSPA8 siRNA (768, 1,112, and 1,509) or nontargeting negative control (NC). Then, the mRNA expression of HSPA8 was performed in HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells. (A) HSPA8-768 at 50 µM, 100 µM, and 200 µM significantly suppressed the mRNA expression of HSPA8. (B) Western blot confirmed the downexpression of HSPA8 in HSPA8-768 (100 µM)-transfected RL-95-2 and HEC-1B cells. *P<0.05 and **P<0.01 vs NC.
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f4-ott-9-2169: Screening of HSPA8 siRNA for knockdown of HSPA8 in RL-95-2 and HEC-1B cells.Notes: RL-95-2 cells were transfected with 50 µM, 100 µM, and 200 µM HSPA8 siRNA (768, 1,112, and 1,509) or nontargeting negative control (NC). Then, the mRNA expression of HSPA8 was performed in HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells. (A) HSPA8-768 at 50 µM, 100 µM, and 200 µM significantly suppressed the mRNA expression of HSPA8. (B) Western blot confirmed the downexpression of HSPA8 in HSPA8-768 (100 µM)-transfected RL-95-2 and HEC-1B cells. *P<0.05 and **P<0.01 vs NC.

Mentions: The role of HSPA8 in endometrial carcinoma was further investigated. HSPA8 siRNA was screened. Transfection of HSPA8 siRNA (768, 1,112, and 1,509) and nontargeting negative control was performed with different concentrations (50 µM, 100 µM, and 200 µM; Figure 4). HSPA8-768 siRNA suppressed the mRNA expression of HSPA8 significantly at 50 µM, 100 µM, and 200 µM (Figure 4A). By transfection with HSPA8-768 siRNA (100 µM), the expression of HSPA8 in both RL-95-2 and HEC-1B cells was significantly downexpressed (Figure 4B). Then, 100 µM of HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells were used in the following experiments.


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)

Screening of HSPA8 siRNA for knockdown of HSPA8 in RL-95-2 and HEC-1B cells.Notes: RL-95-2 cells were transfected with 50 µM, 100 µM, and 200 µM HSPA8 siRNA (768, 1,112, and 1,509) or nontargeting negative control (NC). Then, the mRNA expression of HSPA8 was performed in HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells. (A) HSPA8-768 at 50 µM, 100 µM, and 200 µM significantly suppressed the mRNA expression of HSPA8. (B) Western blot confirmed the downexpression of HSPA8 in HSPA8-768 (100 µM)-transfected RL-95-2 and HEC-1B cells. *P<0.05 and **P<0.01 vs NC.
© Copyright Policy
Related In: Results  -  Collection

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

f4-ott-9-2169: Screening of HSPA8 siRNA for knockdown of HSPA8 in RL-95-2 and HEC-1B cells.Notes: RL-95-2 cells were transfected with 50 µM, 100 µM, and 200 µM HSPA8 siRNA (768, 1,112, and 1,509) or nontargeting negative control (NC). Then, the mRNA expression of HSPA8 was performed in HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells. (A) HSPA8-768 at 50 µM, 100 µM, and 200 µM significantly suppressed the mRNA expression of HSPA8. (B) Western blot confirmed the downexpression of HSPA8 in HSPA8-768 (100 µM)-transfected RL-95-2 and HEC-1B cells. *P<0.05 and **P<0.01 vs NC.
Mentions: The role of HSPA8 in endometrial carcinoma was further investigated. HSPA8 siRNA was screened. Transfection of HSPA8 siRNA (768, 1,112, and 1,509) and nontargeting negative control was performed with different concentrations (50 µM, 100 µM, and 200 µM; Figure 4). HSPA8-768 siRNA suppressed the mRNA expression of HSPA8 significantly at 50 µM, 100 µM, and 200 µM (Figure 4A). By transfection with HSPA8-768 siRNA (100 µM), the expression of HSPA8 in both RL-95-2 and HEC-1B cells was significantly downexpressed (Figure 4B). Then, 100 µM of HSPA8-768 siRNA-transfected RL-95-2 and HEC-1B cells were used in the following experiments.

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