The role of microRNAs in the pathogenesis of recurrent uterine leiomyomas

• Romella A. Karamian
• Irina M. Ordiyants
• Yulia А. Veryaskina
• Dmitry G. Aryutin
• Konstantin V. Rizhenkov

Abstract

The aim of the study was to determine the epigenetic determinants of recurrent uterine leiomyoma at reproductive age patients.

Material and methods. The study included 66 patients who were divided into two groups: I – 42 patients who underwent myomectomy, of which 18 (42.9%) had a history of relapse; II – 24 who underwent hysterectomy, of which 4 (16, 7%) with a history of relapse. The patients underwent treatment and observation in the gynecology department at City Clinical Hospital # 29 named after N.E. Bauman. A section of tissue measuring 5×5 cm from leiomyoma nodes and myometrium was cut out from the surgical material in case of hysterectomy. The cut tissue was frozen at -25 °C. The expression of 21 microRNAs was studied: let7, 20a-3p, 23b, 29b, 146b-5p, 92a-1-5, 93-3p, 100-3p, 10b, 223-3p, 182-5p, 146b-5p, 21-5p, 200a-3p, 30a-3p, 156a-5p, 122-5p, 10a-5p, 15a, 16, 26a. MicroRNA expression levels were measured by real-time PCR using a CFX96 amplifier (Bio-Rad Laboratories, USA). Analysis of the obtained real-time PCR threshold cycle data was carried out using the 2(–∆Ct) method (Livak and Schmittgen, 2001).

Statistical processing of the obtained data was carried out on the basis of the nonparametric Mann–Whitney coefficient using STATISTICA 12.0 software (StatSoft Inc., USA). If p<0.05, the difference was considered statistically significant

Design. The prospective non-randomized longitudinal study was performed.

Results. The groups were comparable in age at menarche and onset of sexual activity, frequency of previous and concomitant extragenital and gynecological diseases. Myomectomy was performed in younger patients 39.6 years old (37.9–41.2) compared to the group of patients with hysterectomy – 46.8 years old (46.0–47.5) with a statistically significant difference p=0.001. When studying the parity of patients in groups I and II, statistically significant differences were also revealed (p=0.021). In reproductive age, patients with infertility underwent myomectomy significantly more often compared to patients with a history of spontaneous childbirth (p=0.032). When comparing the duration of the disease depending on the volume of surgical treatment, no statistically significant differences were found. At the same time, myomectomy was performed on every fourth patient with a uterine size up to 10 weeks of pregnancy (38.1%), every second from 11 to 15 weeks of pregnancy (50.0%) and 11.9% – more than 15 weeks of pregnancy, and in the group of patients operated on in the scope of hysterectomy in 4.2%; 66.7% and 29.2% respectively, p=0,006. When comparing the size of the uterus depending on the volume of surgical treatment, statistically significant differences were obtained (p=0.005).

Every second patient who underwent myomectomy [n=18 (42.9%)] has a history of reccurence of the disease occurred within 1–3 years, and every sixth person who underwent hysterectomy had a history of relapse of the disease [n=4 (16.7%)]. When analyzing the parity of patients with recurrent UL and without relapse, statistically significant differences were obtained (p=0.012). It was found that patients without a history of childbirth have a higher probability of recurrent UL compared to patients with a history of spontaneous childbirth (p=0.023). There was a relatively strong connection between the compared characteristics (Cramer’s V = 0.360). Statistically significant differences were also established in the duration of the disease in the compared groups (with and without relapse, p=0.023).

When comparing the level of microRNA expression in myomatous nodes depending on the volume of surgical treatment (myomectomy or hysterectomy), no statistically significant differences were revealed. However, when comparing the expression of microRNAs in myomatous nodes and myometrium in the material after hysterectomy, statistically significant differences were obtained for the following 8 microRNAs out of 20: let7, 20a, 26a, 29b, 100, 10b, 21-5p, 10a-5p. The expression of Mir Let7, Mir 20a, Mir 100, Mir 10a-5p was one and a half times lower in the myomatous node compared with similar indicators in the myometrium, and Mir 26a, Mir 29b, Mir 10b – two times lower. As for Mir 21-5p, the expression of this microRNA was 1.6 times higher in the myomatous node compared to that in the myometrium.

Conclusion. The identified differences in the expression of 8 microRNAs out of 20 studied: let7, 20a, 26a, 29b, 100, 10b, 21-5p, 10a-5p prove that these nucleotide sequences play a key role in the formation of UL. This provides incentive for further in-depth research in this area. We have not identified a relationship between the level of microRNA expression in myomatous nodes depending on the volume of surgical treatment (myomectomy or hysterectomy) and in the comparison groups depending on the history of relapse. On the contrary, a decrease in the expression levels of Mir Let7, Mir 20a, Mir 100, Mir 10a-5p was detected by one and a half times in the myomatous node, compared with similar indicators in the myometrium, and Mir 26a, Mir 29b, Mir 10b – by a factor of two. As for Mir 21-5p, the expression of this microRNA was 1.6 times higher in the myomatous node compared to that in the myometrium.

Keywords:leiomyoma, recurrent uterine fibroids, microRNA

References

1. Lu T., Rothenberg M. MicroRNA. J Allergy Clin Immunol. 2018; 141 (4): 1202–7. DOI: https://doi.org/10.1016/j.jaci.2017.08.034

2. Camargo A., Remoli B., Portela L., et al. Transcriptomic landscape of male and female reproductive cancers: Similar pathways and molecular signatures predicting response to endocrine therapy. Mol Cell Endocrinol. 2021; 535: 111393. DOI: https://doi.org/10.1016/j.mce.2021.111393

3. Lazzarini R., Caffarini M., Delli Carpini G., et al. From 2646 to 15: differentially regulated microRNNAs between progenitors from normal myometrium and leiomyoma. Am J Obstet Gynecol 2020; 222: 596.e1–9.

4. De Tomi E., Campagnari R., Orlandi E., et al. Upregulation of microRNA-34a-5p, microRNA-20a-3p and microRNA-29a-3p by onconase in A375 melanoma cells correlates with the downregulation of specific onco-proteins. Int J Mol Sci. 2022; 23 (3): 1647. DOI: https://doi.org/10.3390/ijms23031647 Epub 2022 Jan 31.

5. Cao K., Li B., Zhang Y., et al. microRNA-29b restrains cholangiocarcinoma progression by relieving DNMT3B-mediated repression of CDKN2B expression. Aging (Albany NY). 2021; 13 (4): 6055–65. DOI: https://doi.org/10.18632/aging.202549

6. Zhang H., Wang R., Deng Q. microRNA-29b regulates lung cancer progression by downregulating FEM1B and inhibiting the FOX01/AKT pathway. Comput Math Methods Med. 2022; 2022: 3110330. DOI: https://doi.org/10.1155/2022/3110330 Epub 2022 Aug 14.

7. Liu B., Cui Y., Chen W., et al. microRNA-100-3p controls the proliferation, DNA synthesis, and apoptosis of human Sertoli cells by binding to SGK3. Front Cell Dev Biol. 2021; 9: 642916. DOI: https://doi.org/10.3389/fcell.2021.642916 Epub 2021 May 11.

8. Bahena-Ocampo I., Espinosa M., Ceballos-Cancino G., et al. microRNA-10b expression in breast cancer stem cells supports self-renewal through negative PTEN regulation and sustained AKT activation. EMBO Rep. 2016; 17 (5): 648–58. DOI: https://doi.org/10.15252/embr.201540678

9. Liu M., Mo F., Song X., et al. Exosomal microRNA-21-5p is a biomarker for breast cancer diagnosis. PeerJ. 2021; 9: e12147. DOI: https://doi.org/10.7717/peerj.12147 Epub 2021 Sep 17.

10. Liu L., Sun X., Yang C., et al. microRNA -10a-5p restrains the aggressive phenotypes of ovarian cancer cells by inhibiting HOXA1. Kaohsiung J Med Sci. 2021; 37 (4): 276–85. DOI: https://doi.org/10.1002/kjm2.12335

11. Gu Y., Feng X., Jin Y., et al. Upregulation of microRNA-10a-5p promotes tumor progression in cervical cancer by suppressing UBE2I signaling. J Obstet Gynaecol. 2023; 43 (1): 2171283. DOI: https://doi.org/10.1080/01443615.2023.2171283

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