miR-193b-5p and miR-374b-5p Are Aberrantly Expressed in Endometriosis and Suppress Endometrial Cell Migration In Vitro
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Population and Sample Collection
2.2. Ethics
2.3. Cell Lines
2.4. Total RNA Extraction
2.5. Small RNA Library Preparation and Sequencing
2.6. mRNA Library Preparation and Sequencing
2.7. Sequencing Data Processing and Bioinformatic Analysis
2.8. Target Gene Prediction and Pathway Analysis
2.9. Transfection of miRNA Mimics
2.10. qRT-PCR
2.11. Wound Healing Assay
2.12. Transwell Cell Migration Assay
2.13. BrdU cell Proliferation Assay
2.14. Statistical Analysis
3. Results
3.1. DE-miRNAs in Eutopic Endometrium from Women with and Without Endometriosis
3.2. Predicted miRNA Target Genes Are Enriched in Biological Pathways Related to Cell Migration and Proliferation
3.3. Selection of DE-miRNAs for Ex Vivo and In Vitro Studies
3.4. Overexpression of miR-193b-5p and miR-374b-5p Does Not Affect the Proliferation of 12Z and HESC Cells
3.5. Overexpression of miR-193b-5p and miR-374-5p Suppresses Migration of 12Z and HESC Cells
3.6. Identification of Target Genes of miR-193b-5p and miR-374b-5p
3.7. Potential Key miRNA Target Genes Relevant to Cell Migration in Endometriosis
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Cohort 1 | Cohort 2 | ||
---|---|---|---|
Endometriosis (n = 15) | Non-Endometriosis Controls (n = 17) | Endometrioma (n = 10) | |
Age (mean, range), years | 33 (24–43) | 33 (25–38) | 36 (27–42) |
BMI (mean, SD 1), kg/m2 | 22 ± 3.4 | 24 ± 4.7 | 21 ± 2.8 |
Smoker | 1 (6.7%) | 3 (17.6%) | 2 (20.0%) |
Menstrual cycle day 2, mean,(range), days | 8.2 (2–12) | 8.8 (1–13) | 9.9 (4–12) |
Infertility rate | 7 (50%) | 0 | 3 (30%) |
Endometriosis stage | |||
- III | 9 (60%) | na | 7 (70%) |
- IV | 6 (40%) | na | 3 (30%) |
miRNA ID | Fold Change | Adjusted p-Value | |
---|---|---|---|
Upregulated miRNAs | hsa-miR-374b-5p | 3.9 | 0.012 |
hsa-miR-1307-5p | 3.7 | 0.003 | |
hsa-miR-561-5p | 3.6 | <0.000 | |
hsa-miR-144-3p | 2.8 | 0.015 | |
hsa-miR-19a-3p | 2.7 | 0.015 | |
hsa-miR-18b-5p | 2.6 | 0.012 | |
hsa-miR-136-5p | 2.3 | 0.018 | |
hsa-miR-96-5p | 2.2 | 0.020 | |
hsa-miR-101-3p | 2.0 | 0.025 | |
Downregulated miRNAs | hsa-miR-193b-5p | −3.7 | <0.000 |
hsa-miR-374c-3p | −3.6 | 0.006 | |
hsa-miR-320b | −2.5 | 0.005 | |
hsa-miR-320a-3p | −2.2 | <0.000 | |
has-miR-4521 | −2.2 | 0.020 |
Gene Set | Description | Enrichment Score | p-Value |
---|---|---|---|
Path:hsa04380 | Osteoclast differentiation | 17.165 | 3.51× 10−8 |
path:hsa04145 | Phagosome | 17.119 | 3.67 × 10−8 |
path:hsa04514 | Cell adhesion molecules | 16.817 | 4.97 × 10−8 |
path:hsa05150 | Staphylococcus aureus infection | 15.827 | 1.34 × 10−7 |
path:hsa05166 | Human T-cell leukemia virus 1 infection | 13.484 | 1.39 × 10−6 |
path:hsa04924 | Renin secretion | 12.517 | 3.66 × 10−6 |
path:hsa04151 | PI3K-Akt signaling pathway | 12.237 | 4.84 × 10−6 |
path:hsa04640 | Hematopoietic cell lineage | 12.216 | 4.95 × 10−6 |
path:hsa04015 | Rap1 signaling pathway | 10.953 | 1.75 × 10−5 |
path:hsa04662 | B cell receptor signaling pathway | 10.922 | 1.81 × 10−5 |
path:hsa04512 | ECM-receptor interaction | 10.900 | 1.83 × 10−5 |
path:hsa05323 | Rheumatoid arthritis | 10.279 | 3.43 × 10−5 |
path:hsa04713 | Circadian entrainment | 10.020 | 4.42 × 10−5 |
path:hsa04510 | Focal adhesion | 9.901 | 5.01 × 10−5 |
path:hsa05200 | Pathways in cancer | 9.833 | 5.36 × 10−5 |
Gene Symbol | Gene Name | Expression Direction |
---|---|---|
DKK1 | Dickkopf WNT Signaling Pathway Inhibitor 1 | Downregulated in miR-193b-5p transfected 12Z Downregulated in miR-374b-5p transfected 12Z Downregulated in miR-193b-5p transfected HESC Downregulated in miR-374b-5p transfected HESC Downregulated in endometrioma tissue |
MIEN1 | Migration And Invasion Enhancer 1 | Downregulated in endometrioma tissue Downregulated in miR-193b-5p transfected 12Z |
GRB7 | Growth Factor Receptor-Bound Protein 7 | Downregulated in miR-193b-5p transfected 12Z Downregulated in endometrioma tissue |
PIK3R1 | Phosphoinositide-3-Kinase Regulatory Subunit 1 | Downregulated in miR-193b-5p transfected HESC Downregulated in miR-374b-5p transfected HESC Downregulated in endometrioma tissue |
KRT19 | Keratin 19 | Downregulated in miR-193b-5p transfected 12Z Downregulated in miR-374b-5p transfected HESC Downregulated in endometrioma tissue |
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Frisendahl, C.; Tang, Y.; Boggavarapu, N.R.; Peters, M.; Lalitkumar, P.G.; Piltonen, T.T.; Arffman, R.K.; Salumets, A.; Götte, M.; Korsching, E.; et al. miR-193b-5p and miR-374b-5p Are Aberrantly Expressed in Endometriosis and Suppress Endometrial Cell Migration In Vitro. Biomolecules 2024, 14, 1400. https://doi.org/10.3390/biom14111400
Frisendahl C, Tang Y, Boggavarapu NR, Peters M, Lalitkumar PG, Piltonen TT, Arffman RK, Salumets A, Götte M, Korsching E, et al. miR-193b-5p and miR-374b-5p Are Aberrantly Expressed in Endometriosis and Suppress Endometrial Cell Migration In Vitro. Biomolecules. 2024; 14(11):1400. https://doi.org/10.3390/biom14111400
Chicago/Turabian StyleFrisendahl, Caroline, Yiqun Tang, Nageswara Rao Boggavarapu, Maire Peters, Parameswaran Grace Lalitkumar, Terhi T. Piltonen, Riikka K. Arffman, Andres Salumets, Martin Götte, Eberhard Korsching, and et al. 2024. "miR-193b-5p and miR-374b-5p Are Aberrantly Expressed in Endometriosis and Suppress Endometrial Cell Migration In Vitro" Biomolecules 14, no. 11: 1400. https://doi.org/10.3390/biom14111400
APA StyleFrisendahl, C., Tang, Y., Boggavarapu, N. R., Peters, M., Lalitkumar, P. G., Piltonen, T. T., Arffman, R. K., Salumets, A., Götte, M., Korsching, E., & Gemzell-Danielsson, K. (2024). miR-193b-5p and miR-374b-5p Are Aberrantly Expressed in Endometriosis and Suppress Endometrial Cell Migration In Vitro. Biomolecules, 14(11), 1400. https://doi.org/10.3390/biom14111400