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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

The proliferation, cell cycle, and apoptosis of HSPA8 siRNA-transfected RL-95-2 and HEC-1B cells.Notes: After transfection for 48 hours, cell proliferation was detected by MTT assay (A); cell apoptosis was determined using the Annexin V-FITC/PI flow cytometry (B), and proportion of apoptosis cells was measured (C); cell cycle distribution in each group was measured by flow cytometry (D) and the cell cycle phase is shown in a bar graph (E) form with the G0/G1, S, and G2/M phases. *P<0.05, **P<0.01, and HSPA8 siRNA vs NC.Abbreviations: PI, propidium iodide; NC, negative control; h, hours; FITC, fluorescein isothiocyanate.
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f5-ott-9-2169: The proliferation, cell cycle, and apoptosis of HSPA8 siRNA-transfected RL-95-2 and HEC-1B cells.Notes: After transfection for 48 hours, cell proliferation was detected by MTT assay (A); cell apoptosis was determined using the Annexin V-FITC/PI flow cytometry (B), and proportion of apoptosis cells was measured (C); cell cycle distribution in each group was measured by flow cytometry (D) and the cell cycle phase is shown in a bar graph (E) form with the G0/G1, S, and G2/M phases. *P<0.05, **P<0.01, and HSPA8 siRNA vs NC.Abbreviations: PI, propidium iodide; NC, negative control; h, hours; FITC, fluorescein isothiocyanate.

Mentions: The knockdown of HSPA8 significantly reduced the cell proliferation at 48 hours (Figure 5A). Cell apoptosis including both early apoptosis and later apoptosis was promoted by HSPA8 siRNA (Figure 5B and C). In addition, the knockdown of HSPA8 significantly reduced the cells in S phases (Figure 5D and E), suggesting that HSPA8 plays an important role in the development of endometrial carcinoma. HSPA8 is a candidate for early diagnosis and therapy biomarker for endometrial carcinoma.


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 proliferation, cell cycle, and apoptosis of HSPA8 siRNA-transfected RL-95-2 and HEC-1B cells.Notes: After transfection for 48 hours, cell proliferation was detected by MTT assay (A); cell apoptosis was determined using the Annexin V-FITC/PI flow cytometry (B), and proportion of apoptosis cells was measured (C); cell cycle distribution in each group was measured by flow cytometry (D) and the cell cycle phase is shown in a bar graph (E) form with the G0/G1, S, and G2/M phases. *P<0.05, **P<0.01, and HSPA8 siRNA vs NC.Abbreviations: PI, propidium iodide; NC, negative control; h, hours; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

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

f5-ott-9-2169: The proliferation, cell cycle, and apoptosis of HSPA8 siRNA-transfected RL-95-2 and HEC-1B cells.Notes: After transfection for 48 hours, cell proliferation was detected by MTT assay (A); cell apoptosis was determined using the Annexin V-FITC/PI flow cytometry (B), and proportion of apoptosis cells was measured (C); cell cycle distribution in each group was measured by flow cytometry (D) and the cell cycle phase is shown in a bar graph (E) form with the G0/G1, S, and G2/M phases. *P<0.05, **P<0.01, and HSPA8 siRNA vs NC.Abbreviations: PI, propidium iodide; NC, negative control; h, hours; FITC, fluorescein isothiocyanate.
Mentions: The knockdown of HSPA8 significantly reduced the cell proliferation at 48 hours (Figure 5A). Cell apoptosis including both early apoptosis and later apoptosis was promoted by HSPA8 siRNA (Figure 5B and C). In addition, the knockdown of HSPA8 significantly reduced the cells in S phases (Figure 5D and E), suggesting that HSPA8 plays an important role in the development of endometrial carcinoma. HSPA8 is a candidate for early diagnosis and therapy biomarker for endometrial carcinoma.

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