effect of basigin (cd147) on heparin-binding egf-like growth factor and matrix metalloproteinase...
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Effect of basigin (CD147) on heparin-binding EGF-like growth factor and matrix metalloproteinase expression in human cytotrophoblast cells
AuthorsInstitutions
Background - HB-EGF, MMPs
● Trophoblast cell survival is dependent on matrix metaloproteinase (MMP) mediated shedding HB-EGF
● Basigin is an Ig superfamily cell surface glycoprotein with multiple functions, including induction of MMPs
Mol Reprod Dev 2009, 76(12): 1116-1127
Background - What can we learn from Basigin null mice ?● Severely impaired implantation
– Normal embryos, but <8% achieved implantation
● Females are capable of supporting Basigin wildtype embryos, but with severely reduced efficiency
● Males experience spermatogenesis arrest at M1 metaphase
● Impaired neutrophil migration into renal tissue after ischemia/reperfusion (renal injury)
● Deficient MCT1, MCT3, MCT4 in retina
Figure 1. Basigin expression in implantation sites of the mouse uterus on d 5 (A), d 6 (B), d 7 (C), and d 8 (D) of pregnancy
Endocrinology 2009, 150: 966-976
Background - Basigin in Pregnancy
Figure 1. Expression profiles of MMP-2, MMP-3, MMP-9, and TIMP-3 mRNA during in vitro decidualization.
Figure 2. Treatment of uterine stromal cells with rBSG increases MMP-3 and MMP-9 mRNA levels without affecting MMP-2 or TIMP-3 expressionEndocrinology 2009; 150: 966-976
Mouse Uterus rBSG
ExpressionProfile
Objective: To determine the effect of basigin on HB-EGF and MMP expression in human first trimester cytotrophoblast cells.
BSG?
?
Mol Reprod Dev 2009, 76(12): 1116-1127
Methods - rBSG Treatment, ELISA, and Gene Expression Profiling
rBSG
ELISA: HB-EGF, MMPsprotein quantification using cell medium and lysates
Gene Expression Profiling
http://www.allelebiotech.cn/mgr/eWebEditor/UploadFile/20071018225116615.jpg
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rBSG (_g/ml)
MMP-2 protein
concentration (ng/ml)
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rBSG (_g/ml)
HB-EGF protein concentration (ng/ml)
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rBSG (_g/ml)
MMP-9 protein
concentration (ng/ml)
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Figure 1. rBSG does not stimulate (A) HB-EGF, (B) MMP-2 and (C) MMP-9 protein expression from HTR cells. Cells were treated with rBSG at 0 (control), 0.1, 1, and 5 µg/ml for 24 h as described, and the cell lysates and MMP-2 were collected. Data expressed as mean from separate experiments.
Results - ELISA
HB-EGF
MMP-2
MMP-9
< - 4 > 41FC
Con
trol
BS
G 0
.5
gB
SG
1.0
g
BS
G 5
.0
g
Results - Gene Expression Profile
Figure 2. Heatmap and hierarchical clustering of 2242 features altered by 2-fold.
MMP-19
● HB-EGF, MMP-2 and MMP-9 protein levels were not increased in HTR cells following 24 hour treatment with rBSG (Figure 1)– Non-statistically significant increases (1.1 to 1.7 fold) in
MMP-9● Global gene expression and higherarchial clustering
revealed 2,242 features to be altered more than 2 fold (Figure 2)– MMP-19 was found to be altered by more than 2 fold
● Protein analysis revealed 60 genes to have more than 10 protein-protein interactions
Results - Summary
● Effect of Basigin on human trophoblast cells– HB-EGF expression - No Effect– Protein and gene expression - Increased MMP-19
● Further research is required to better understand the role of MMP-19 in HTR cells
Conclusions
Thank you for your attention, I am now ready for questions!
This work was supported by a VARI-GRMERC Medical Student Summer Research Internship (DJH) and National Institutes of Health Grant U54 HD40093 (REL).
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