Analysis of Reciprocally Dysregulated miRNAs in Eutopic Endometrium Is a Promising Approach for Low Invasive Diagnostics of Adenomyosis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethical Approval and Study Design
2.2. Participants
2.3. RNA Sampling
2.4. miRNA Expression Profiling Using Exiqon microRNA PCR Panel
2.5. Quantitative Real-Time RT-PCR (qPCR)
2.6. Calculation and Statistics
3. Results
3.1. Preliminary miRNA Profiling
3.2. Creation and Validation of ttRT-qPCR Systems for Individual miRNA Analysis
3.3. Analysis of Selected miRNAs by ttRT-qPCR Systems
3.4. Identification of Reciprocally Dysregulated miRNA Pairs and Analysis of Their Diagnostic Potency
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Location (Uteri Wall) | Anterior = 4 | Posterior = 22 | Mixed/Other = 7 | |
---|---|---|---|---|
Differentiation | Focal = 10 | Diffuse = 17 | Adenomyoma = 6 | |
Cystic | Yes = 28 | No = 5 | ||
Uterine layer involvement | Type 1–2 = 19 | Type 2–3 = 8 | Type 1–3 = 6 | |
Extent of adenomyosis | Moderate = 21 | Severe = 12 | ||
Size of lesions | 3–6 cm = 8 | 7–9 cm = 22 | 10 and >=3 |
miRNA | Method Sensitivity Limit for Synthetic miRNA | Results of miRNA Analysis in Biological Samples(Ct Value Range and Median) | |
---|---|---|---|
Plateau of Ct Value | Minimal Concentration (Molecules/Reaction) | ||
-10a | 36.3 | 104 | 29.32–33.41 (31.80) |
-10b | 34.7 | 105 | 28.12–31.97 (30.60) |
-31 | 30.4 | 105 | 26.03–28.78 (27.45) |
-181b | 29.5 | 107 | 27.03–28.03 (27.45) |
-191 | 31.4 | 105 | 21.42–25.44 (23.67) |
-195 | 32.5 | 105 | 25.31–29.12 (27.31) |
-200b | 33.4 | 105 | 24.11–25.25 (26.87) |
-200c | 33.7 | 105 | 25.18–30.22 (27.53) |
-221 | 32.6 | 106 | 29.55–31.88 (30.88) |
miRNA- | -10a | -10b | -31 | -181b | -191 | -195 | -200b | -200c | -221 | |
---|---|---|---|---|---|---|---|---|---|---|
CNT | Mean | 0.23 | 1.72 | 3.49 | 2.38 | 20.04 | 7.56 | 3.48 | 4.57 | 0.43 |
SEM | 0.06 | 0.51 | 0.76 | 0.71 | 6.18 | 2.77 | 0.93 | 1.17 | 0.14 | |
AM | Mean | 0.24 | 0.19 | 3.05 | 3.63 | 41.34 | 8.48 | 5.71 | 1.73 | 0.22 |
SEM | 0.06 | 0.04 | 0.55 | 0.75 | 9.56 | 2.16 | 3.02 | 0.32 | 0.07 | |
Mann–Whitney | NS | 0.0017 | 0.9490 | 0.0680 | 0.0011 | NS | NS | 0.0200 | NS |
miR-191/-10b | miR-191/-31 | miR-181b/-10b | miR-181b/200c | miR-191/-200c | |
---|---|---|---|---|---|
Diagnostic potency (AUC) | 0.75 | 0.76 | 0.77 | 0.75 | 0.74 |
Sensitivity (%) | 70.97 | 73.33 | 61.29 | 74.19 | 64.52 |
Specificity (%) | 79.31 | 79.31 | 72.41 | 75.86 | 86.21 |
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Borisov, E.; Knyazeva, M.; Novak, V.; Zabegina, L.; Prisyazhnaya, T.; Karizkiy, A.; Berlev, I.; Malek, A. Analysis of Reciprocally Dysregulated miRNAs in Eutopic Endometrium Is a Promising Approach for Low Invasive Diagnostics of Adenomyosis. Diagnostics 2020, 10, 782. https://doi.org/10.3390/diagnostics10100782
Borisov E, Knyazeva M, Novak V, Zabegina L, Prisyazhnaya T, Karizkiy A, Berlev I, Malek A. Analysis of Reciprocally Dysregulated miRNAs in Eutopic Endometrium Is a Promising Approach for Low Invasive Diagnostics of Adenomyosis. Diagnostics. 2020; 10(10):782. https://doi.org/10.3390/diagnostics10100782
Chicago/Turabian StyleBorisov, Evgeny, Margarita Knyazeva, Veronika Novak, Lidia Zabegina, Tatyana Prisyazhnaya, Aleksey Karizkiy, Igor Berlev, and Anastasia Malek. 2020. "Analysis of Reciprocally Dysregulated miRNAs in Eutopic Endometrium Is a Promising Approach for Low Invasive Diagnostics of Adenomyosis" Diagnostics 10, no. 10: 782. https://doi.org/10.3390/diagnostics10100782