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. 2024 Apr;14(4):e1672.
doi: 10.1002/ctm2.1672.

Synergistic inhibition of progesterone receptor-A/B signalling by simvastatin and mifepristone in human uterine leiomyomas

Sadia Afrin  1 , Gregory W Kirschen  1 , Mariko Miyashita-Ishiwata  1 , Malak El Sabeh  2 , Mostafa A Borahay  1
Affiliations

Synergistic inhibition of progesterone receptor-A/B signalling by simvastatin and mifepristone in human uterine leiomyomas

Sadia Afrin et al. Clin Transl Med. 2024 Apr.
No abstract available

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Suppression of progesterone receptor (PR) expression in simvastatin‐treated clinical tissue. (A) Diagram of clinical trial design of simvastatin (SIM) versus placebo for treatment of leiomyomas. Patients with uterine fibroids were recruited for treatment with simvastatin (40 mg daily) or a placebo (starch 1500 encapsulated) for a total of 12 weeks. At the end of the treatment, fibroid samples were collected after the surgery to evaluate the effects of simvastatin. (B) Tissue was fixed with a 10% buffered formalin solution for 24 h and kept in 70% ethanol at 4°C. Tissue was then stained with pPR‐A/B antibody (green fluorescence) and 4'‐6‐Diamidino‐2‐phenylindole (DAPI) (blue fluorescence). Scale bars, 50 µm. (C) The optical density was measured and quantified in 15 images using ImageJ, using 10 histologically similar fields randomly selected from each slide. * p < .05 versus the placebo.
FIGURE 2
FIGURE 2
Simvastatin treatment suppressed progesterone receptor (PR)‐A/B expression in a xenograft mouse model and primary cells. (A) 6‐week old female immunodeficient NOG (NOD/Shi‐scid/IL‐2Rγnull) mice (n = 12 control, 11 simvastatin [SIM]) were implanted with two human‐derived fibroid tumours each and assigned to vehicle control or SIM (20 µg/g body weight/day for 28 consecutive days). We stained fibroid tumours for PR and observed significantly decreased expression in the SIM group (p = .036). The data presented here are previously unpublished PR expressions from our previously reported xenograft leiomyoma mouse model (cartoon adapted)., (B) Immunocytochemistry staining was performed on primary leiomyoma cells to confirm the cellular localization of pPR‐A/B (green fluorescence) and 4'‐6‐Diamidino‐2‐phenylindole (DAPI) (blue fluorescence). (C) total and (D) phosphorated PR‐A/B expression after 48 h SIM treatment (WB). Β‐actin was used as a loading control. Scale bar, 50 µm. Each condition was repeated in triplicate. Each graph represents the experimental data with standard error of the mean (SEM) of three independent experiments with similar results. *, p  <  .05 versus the control; #, p  <  .05 versus P4.
FIGURE 3
FIGURE 3
Inhibition of progesterone downstream signalling and transcription by simvastatin. The expression of progesterone receptor (PR) downstream signalling (A) pERK/ERK, (B) p‐JNK/JNK, (C) pAKT/AKT, (D) pmTOR1/mTOR1 and (E) PGMRC1 were assessed in primary leiomyoma cells after simvastatin (SIM) treatment. Protein expressions were assessed using the Western blot (WB) assay. β‐actin was used as a loading control. Dimethyl sulfoxide (DMSO) was used as vehicle control. (F) Transcriptional activity of progesterone in human immortalized leiomyoma (HuLM) cells after simvastatin treatment. Leiomyoma cells were co‐transfected with plasmids of PRE‐luc (500 ng/well) and PR‐B (50 ng/well) for 8 h. Cells were treated with P4 (100 nM) and SIM (0.001–10 µM) alone and combination for 48 h. Cells were harvested and assayed for luciferase activity. The data is presented as relative light units (RLU) defined as the luciferase output normalized to protein content. The fold changes were calculated by normalization with luciferase value with no P4 treatment. Each condition was repeated in triplicate. Each graph represents the experimental data with SEM of three independent experiments with similar results. *, p  <  .05 versus the control; #, p  <  .05 versus P4.
FIGURE 4
FIGURE 4
Simvastatin and mifepristone synergistically inhibit proliferation and downstream targets. The expressions of (A) PCNA, (B) pERK1/2/ ERK1/2, (C) pJNK1/2/ JNK1/2 and (D) pAKT/ AKT were assessed in primary leiomyoma cells after treatment with P4 (100 nM), simvastatin (SIM) (1 µM), Mifepristone (MIF,1 µM) alone or combination for 48 h. Protein expressions were assessed using WB assay. β‐actin was used as a loading control. Dimethyl sulfoxide (DMSO) was used as vehicle control. Each condition was repeated in triplicate. Each graph represents the experimental data with SEM of three independent experiments with similar results. *, p  <  .05 versus the control; #, p  <  .05 versus P4.
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References

    1. Islam MS, Ciavattini A, Petraglia F, Castellucci M, Ciarmela P. Extracellular matrix in uterine leiomyoma pathogenesis: a potential target for future therapeutics. Hum Reprod Update. 2018;24(1):59‐85. - PubMed
    1. Enazy SAl, Kirschen GW, Vincent K, et al. PEGylated polymeric nanoparticles loaded with 2‐methoxyestradiol for the treatment of uterine leiomyoma in a patient‐derived xenograft mouse model. J Pharm Sci. 2023;112(9):2552‐2560. - PMC - PubMed
    1. Talaulikar VS, Manyonda I. Progesterone and progesterone receptor modulators in the management of symptomatic uterine fibroids. Eur J Obstet Gynecol Reprod Biol. 2012;165(2):135‐140. - PubMed
    1. Afrin S, Islam MdS, Patzkowsky K, et al. Simvastatin ameliorates altered mechanotransduction in uterine leiomyoma cells. Am J Obstet Gynecol. 2020;223(5):733.e1‐733.e14. - PMC - PubMed
    1. Shimomura Y, Matsuo H, Samoto T, Maruo T. Up‐regulation by progesterone of proliferating cell nuclear antigen and epidermal growth factor expression in human uterine leiomyoma. J Clin Endocrinol Metab. 1998;83(6):2192‐2198. - PubMed

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