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Quantitative proteomic analysis of serum from pregnant women carrying a fetus with conotruncal heart defect using isobaric tags for relative and absolute quantitation (iTRAQ) labeling.

Zhang Y, Kang Y, Zhou Q, Zhou J, Wang H, Jin H, Liu X, Ma D, Li X - PLoS ONE (2014)

Bottom Line: A total of 105 unique proteins present in the three groups were identified, and relative expression data were obtained for 92 of them with high confidence by employing the iTRAQ-based experiments.The downregulation of gelsolin in maternal serum of fetus with CTD was further verified by Western blot.The identification of differentially expressed protein gelsolin in the serum of the pregnant women carrying a CTD fetus by using proteomic technology may be able to serve as a foundation to further explore the biomarker for detection of CTD fetus from the maternal serum.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Hospital, Fudan University, Shanghai, China.

ABSTRACT

Objective: To identify differentially expressed proteins from serum of pregnant women carrying a conotruncal heart defects (CTD) fetus, using proteomic analysis.

Methods: The study was conducted using a nested case-control design. The 5473 maternal serum samples were collected at 14-18 weeks of gestation. The serum from 9 pregnant women carrying a CTD fetus, 10 with another CHD (ACHD) fetus, and 11 with a normal fetus were selected from the above samples, and analyzed by using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry(2D LC-MS/MS). The differentially expressed proteins identified by iTRAQ were further validated with Western blot.

Results: A total of 105 unique proteins present in the three groups were identified, and relative expression data were obtained for 92 of them with high confidence by employing the iTRAQ-based experiments. The downregulation of gelsolin in maternal serum of fetus with CTD was further verified by Western blot.

Conclusions: The identification of differentially expressed protein gelsolin in the serum of the pregnant women carrying a CTD fetus by using proteomic technology may be able to serve as a foundation to further explore the biomarker for detection of CTD fetus from the maternal serum.

No MeSH data available.


Related in: MedlinePlus

Western blot and IHC analyses confirmed the relative expression of GSN in fetal heart tissues with CTD.(A) Western blot analysis examined the relative expression of GSN, and confirmed that the GSN protein was downregulated in heart tissues of CTD fetuses compared with normal controls. n = 4, respectively. *p = 0.004. (B) IHC demonstrated the dark brown immunostaining was weaker in the CTD heart tissues than the normal controls (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks). n = 4, respectively. Magnification was set at 400×.
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pone-0111645-g003: Western blot and IHC analyses confirmed the relative expression of GSN in fetal heart tissues with CTD.(A) Western blot analysis examined the relative expression of GSN, and confirmed that the GSN protein was downregulated in heart tissues of CTD fetuses compared with normal controls. n = 4, respectively. *p = 0.004. (B) IHC demonstrated the dark brown immunostaining was weaker in the CTD heart tissues than the normal controls (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks). n = 4, respectively. Magnification was set at 400×.

Mentions: Having validated that GSN protein was downregulated in maternal serum with a CTD fetus, we also examined the expression of GSN in 4 gestation week-paired CTD fetal hearts (23–25 gestational weeks), and 4 normal controls by Western blot and IHC. Figure 3A demonstrates that the GSN expression in heart tissues of fetuses with CTD in comparison with normal tissue, detected by Western blot. The expression of GSN was downregulated in CTD fetuses in comparison with normal controls. Figure 3B shows the expression level and localization of GSN in heart tissues of the representative cases by IHC. GSN protein is expressed in the cytoplasm of myocardiocytes. The dark brown immunostaining is weaker in the CTD heart tissues than the normal controls, further verifying the downregulation of GSN in CTD fetus' heart (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks).


Quantitative proteomic analysis of serum from pregnant women carrying a fetus with conotruncal heart defect using isobaric tags for relative and absolute quantitation (iTRAQ) labeling.

Zhang Y, Kang Y, Zhou Q, Zhou J, Wang H, Jin H, Liu X, Ma D, Li X - PLoS ONE (2014)

Western blot and IHC analyses confirmed the relative expression of GSN in fetal heart tissues with CTD.(A) Western blot analysis examined the relative expression of GSN, and confirmed that the GSN protein was downregulated in heart tissues of CTD fetuses compared with normal controls. n = 4, respectively. *p = 0.004. (B) IHC demonstrated the dark brown immunostaining was weaker in the CTD heart tissues than the normal controls (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks). n = 4, respectively. Magnification was set at 400×.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230941&req=5

pone-0111645-g003: Western blot and IHC analyses confirmed the relative expression of GSN in fetal heart tissues with CTD.(A) Western blot analysis examined the relative expression of GSN, and confirmed that the GSN protein was downregulated in heart tissues of CTD fetuses compared with normal controls. n = 4, respectively. *p = 0.004. (B) IHC demonstrated the dark brown immunostaining was weaker in the CTD heart tissues than the normal controls (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks). n = 4, respectively. Magnification was set at 400×.
Mentions: Having validated that GSN protein was downregulated in maternal serum with a CTD fetus, we also examined the expression of GSN in 4 gestation week-paired CTD fetal hearts (23–25 gestational weeks), and 4 normal controls by Western blot and IHC. Figure 3A demonstrates that the GSN expression in heart tissues of fetuses with CTD in comparison with normal tissue, detected by Western blot. The expression of GSN was downregulated in CTD fetuses in comparison with normal controls. Figure 3B shows the expression level and localization of GSN in heart tissues of the representative cases by IHC. GSN protein is expressed in the cytoplasm of myocardiocytes. The dark brown immunostaining is weaker in the CTD heart tissues than the normal controls, further verifying the downregulation of GSN in CTD fetus' heart (i, normal control, 23 gestational weeks, ii, CTD 23 gestational weeks; iii normal control, 25 gestational weeks, iv, CTD 25 gestational weeks).

Bottom Line: A total of 105 unique proteins present in the three groups were identified, and relative expression data were obtained for 92 of them with high confidence by employing the iTRAQ-based experiments.The downregulation of gelsolin in maternal serum of fetus with CTD was further verified by Western blot.The identification of differentially expressed protein gelsolin in the serum of the pregnant women carrying a CTD fetus by using proteomic technology may be able to serve as a foundation to further explore the biomarker for detection of CTD fetus from the maternal serum.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Hospital, Fudan University, Shanghai, China.

ABSTRACT

Objective: To identify differentially expressed proteins from serum of pregnant women carrying a conotruncal heart defects (CTD) fetus, using proteomic analysis.

Methods: The study was conducted using a nested case-control design. The 5473 maternal serum samples were collected at 14-18 weeks of gestation. The serum from 9 pregnant women carrying a CTD fetus, 10 with another CHD (ACHD) fetus, and 11 with a normal fetus were selected from the above samples, and analyzed by using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry(2D LC-MS/MS). The differentially expressed proteins identified by iTRAQ were further validated with Western blot.

Results: A total of 105 unique proteins present in the three groups were identified, and relative expression data were obtained for 92 of them with high confidence by employing the iTRAQ-based experiments. The downregulation of gelsolin in maternal serum of fetus with CTD was further verified by Western blot.

Conclusions: The identification of differentially expressed protein gelsolin in the serum of the pregnant women carrying a CTD fetus by using proteomic technology may be able to serve as a foundation to further explore the biomarker for detection of CTD fetus from the maternal serum.

No MeSH data available.


Related in: MedlinePlus