Fractalkine Regulates HEC-1A/JEG-3 Interaction by Influencing the Expression of Implantation-Related Genes in an In Vitro Co-Culture Model
Abstract
:1. Introduction
2. Results
2.1. Effect of Fractalkine on the Viability of JEG-3 Cells in Mono- and Co-Cultures
2.2. Fractalkine Changes the Activation of ERK1/2, p38, JNK and AKT Signalling Pathways in Mono- and Co-Cultured JEG-3 Cells
2.3. Fractalkine Exerts Different Effects on the mRNA Expression of Proliferation, Differentiation and Invasion Regulating Genes in Mono- and Co-Cultured JEG-3 Cells
2.4. Western Blot Analysis of the Implantation-Related Genes Reveals Alterations Between Fractalkine Treated Mono- and Co-Cultured JEG-3 Cells
2.5. The Presence of HEC-1A Cells Contributes to the Action of Fractalkine on JEG-3 Cells by Changing the Expressions of Activin, Follistatin and BMP2
3. Discussion
4. Materials and Methods
4.1. Cell Cultures and Treatments
4.2. Cell Viability Assay
4.3. Real-Time PCR
4.4. Immunoblotting
4.5. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
AcvR | activin receptor |
AKT | protein kinase B |
BMP2 | bone morphogenetic protein 2 |
BMPR | bone morphogenetic protein receptor |
CNS | central nervous system |
CX3CL1 | fractalkine |
CX3CR1 | fractalkine receptor |
ERK | extracellular signal-regulated protein kinase |
FKN | fractalkine |
JNK | c-Jun N-terminal kinase |
MAPK | mitogen-activated protein kinase |
MMP | matrix metalloproteinase |
NFκB | nuclear factor kappa-light-chain-enhancer of activated B cells |
PI3K | phosphatidylinositol 3 kinase |
PKC | protein kinase C |
PLC | phospholipase C |
PR | progesterone receptor |
SMAD2/3 | homologues of the Drosophila protein, mothers against decapentaplegic (Mad) and the caenorhabtidis elegans protein (Sma) |
SRC-1 | steroid receptor coactivator-1 |
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Primer | Sequence 5′ → 3′ |
---|---|
Progesterone receptor A/B forward | CCAAAGGCCGCAAATTCT |
Progesterone receptor A/B reverse | TGAGGTCAGAAAGGTCATCG |
Fractalkine receptor forward | CCATTAGTCTGGGCGTCTGG |
Fractalkine receptor reverse | GTCACCCAGACACTCGTTGT |
Activin receptor 1B forward | CGTTTGCCGTCTTTCTTATC |
Activin receptor 1B reverse | ACCAGTTTGATTGGTTCTGT |
MMP2 forward | GTCGCCCATCATCAAGTT |
MMP2 reverse | GCATCTTCTTTAGTGTGTCCT |
MMP9 forward | CGGACCAAGGATACAGTTTG |
MMP9 reverse | AAGCGGTACATAGGGTACAT |
SRC-1 forward | AGACCCAACCTTTATTCCCA |
SRC-1 reverse | GGTGTTACTTGAACAGGCAT |
Activin A forward | GAACTTATGGAGCAGACCTC |
Activin A reverse | GGACTTTTAGGAAGAGCCAG |
Follistatin forward | CAAAGCAAAGTCCTGTGAAG |
Follistatin reverse | CCTCTCCCAACCTTGAAATC |
BMP2 forward | TAAGTTCTATCCCCACGGAG |
BMP2 reverse | AGCATCTTGCATCTGTTCTC |
β-actin forward | AGAAAATCTGGCACCACACC |
β-actin reverse | GGGGTGTTGAAGGTCTCAAA |
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Pap, R.; Montskó, G.; Jánosa, G.; Sipos, K.; Kovács, G.L.; Pandur, E. Fractalkine Regulates HEC-1A/JEG-3 Interaction by Influencing the Expression of Implantation-Related Genes in an In Vitro Co-Culture Model. Int. J. Mol. Sci. 2020, 21, 3175. https://doi.org/10.3390/ijms21093175
Pap R, Montskó G, Jánosa G, Sipos K, Kovács GL, Pandur E. Fractalkine Regulates HEC-1A/JEG-3 Interaction by Influencing the Expression of Implantation-Related Genes in an In Vitro Co-Culture Model. International Journal of Molecular Sciences. 2020; 21(9):3175. https://doi.org/10.3390/ijms21093175
Chicago/Turabian StylePap, Ramóna, Gergely Montskó, Gergely Jánosa, Katalin Sipos, Gábor L. Kovács, and Edina Pandur. 2020. "Fractalkine Regulates HEC-1A/JEG-3 Interaction by Influencing the Expression of Implantation-Related Genes in an In Vitro Co-Culture Model" International Journal of Molecular Sciences 21, no. 9: 3175. https://doi.org/10.3390/ijms21093175