Breast Cancer Resistance Protein (BCRP/ABCG2) Inhibits Extra Villous Trophoblast Migration: The Impact of Bacterial and Viral Infection
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethical Approval
2.2. Human Placental Explant Culture
2.3. Immunohistochemistry
2.4. Cell Line and Culture
2.5. Quantitative Real Time PCR (qPCR)
2.6. Immunoblotting
2.7. Enzyme-Linked Immunosorbent Assay (ELISA)
2.8. Transient Transfection with siRNA Oligonucleotides
2.9. Cell Migration Assay
2.10. Cell Proliferation Assay
2.11. Statistical Analysis
3. Results
3.1. Expression of BCRP Is Enriched in Column Cytotrophoblasts (CCT) and Extravillous Trophoblasts (EVTs) in Placental Explants
3.2. ABCG2 Knockdown or Functional Inhibition of BCRP Stimulates EVT Cell Migration (HTR8/SVneo Cells)
3.3. Bacterial and Viral Infection Mimics Activate a Pro-Inflammatory Response in EVTs and Impair ABCG2 and BCRP Expression (HTR8/SVneo Cells)
3.4. LPS and ssRNA Exposure Increases EVT Migration (HTR8/SVneo Cells)
3.5. ABCG2 Knockdown Stimulates EVT Cell Migration in Placental Explants
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Primer Name | Sequence | Reference |
---|---|---|
ABCG2 | Forward: 5′-TGGAATCCAGAACAGAGCTGGGGT-3′ | Lye et al., 2018 |
Reverse: 5′-AGAGTTCCACGGCTGAAACACTGC-3′ | ||
IL-6 | Forward: 5′-TGCAGAAAAAGGCAAAGAAT-3′ | Potter et al., 2015 |
Reverse: 5′-CTGACCAGAAGAAGGAATGC-3′ | ||
IL-8 | Forward: 5′-TGGGAACAAGAGGGCATCTG-3′ | Lye et al., 2015 |
Reverse: 5′-CCACCACTGCATCAAATTCATG-3′ | ||
CCL2 | Forward: 5′-TTCATTCCCCAAGGGCTCGCTCA-3′ | Lye et al., 2015 |
Reverse: 5′-AGCACAGATCTCCTTGGCCACAA-3′ | ||
TLR-4 | Forward: 5′-ATTTGTCTCCACAGCCACCA-3′ | Lye et al., 2015 |
Reverse: 5′-ACAGGAAACCCCATCCAGAG-3′ | ||
HPRT | Forward: 5′-TGA CAC TGG CAA AAC AATGCA-3′ | Drewlo et al., 2012 |
Reverse: 5′-GGT CCT TTTCAC CAG CAA GCT-3′ | ||
TBP | Forward: 5′-TGC ACA GGA GCC AAG AGT GAA-3′ | Drewlo et al., 2012 |
Reverse: 5′-CAC ATC ACA GCT CCC CAC CA-3′ |
Number | Sequence | |
---|---|---|
1 | Antisense | AUAACCAGCUGAUUCAAAGUAUCCC |
Sense | GGGAUACUUUGAAUCAGC UGGUUAU | |
2 | Antisense | UAAUGAUGUCCAAGAAGAAGUCUGC |
Sense | CAGACUUCUUCUUGGACAUCAUUA | |
3 | Antisense | GGAGGCAAAUCUUCGUUAUTT |
Sense | AUAACGAAGAUUUGCCUCCTT | |
Negative Control | Antisense | GCGACGAUCUGCCUAAGAUdTdT |
Sense | AUCUUAGGCAGAUCGUCGCdTdT |
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Share and Cite
Lye, P.; Bloise, E.; Nadeem, L.; Peng, C.; Gibb, W.; Ortiga-Carvalho, T.M.; Lye, S.J.; Matthews, S.G. Breast Cancer Resistance Protein (BCRP/ABCG2) Inhibits Extra Villous Trophoblast Migration: The Impact of Bacterial and Viral Infection. Cells 2019, 8, 1150. https://doi.org/10.3390/cells8101150
Lye P, Bloise E, Nadeem L, Peng C, Gibb W, Ortiga-Carvalho TM, Lye SJ, Matthews SG. Breast Cancer Resistance Protein (BCRP/ABCG2) Inhibits Extra Villous Trophoblast Migration: The Impact of Bacterial and Viral Infection. Cells. 2019; 8(10):1150. https://doi.org/10.3390/cells8101150
Chicago/Turabian StyleLye, Phetcharawan, Enrrico Bloise, Lubna Nadeem, Chun Peng, William Gibb, Tania M. Ortiga-Carvalho, Stephen J. Lye, and Stephen G. Matthews. 2019. "Breast Cancer Resistance Protein (BCRP/ABCG2) Inhibits Extra Villous Trophoblast Migration: The Impact of Bacterial and Viral Infection" Cells 8, no. 10: 1150. https://doi.org/10.3390/cells8101150