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Inhibition of tumor cell proliferation in human uterine leiomyomas by vitamin D via Wnt/β-catenin pathway

      Objective

      To assess the effect of vitamin D (VitD) on human uterine leiomyomas through Wnt/β-catenin pathway inhibition, apoptosis induction, and cell growth arrest.

      Design

      A prospective study comparing leiomyoma vs. myometrium tissues. Paired design study comparing human uterine leiomyoma primary (HULP) cells treated with or without VitD.

      Setting

      University hospital.

      Patient(s)

      Human uterine leiomyoma and myometrium were collected from women (aged 35−52 years) without hormonal treatment.

      Intervention(s)

      Samples were collected from women undergoing surgery due to symptomatic uterine leiomyoma pathology.

      Main Outcome Measure(s)

      Uterine leiomyoma and myometrium tissues were analyzed by western blot (WB) to determine proliferation, Wnt/β-catenin, and apoptosis pathways. HULP cells were used to study VitD effect in cell proliferation (WB), cell cycle (flow cytometry), Wnt/β-catenin and apoptosis genes (polymerase chain reaction arrays), Wnt-related proteins (protein array), and apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling [TUNEL] assay).

      Results

      Human leiomyoma tissues compared with matched myometrium showed higher proliferation (fold change = 8.16; P=.0006) and altered Wnt/β-catenin pathway (fold change = 5.5; P<.0001), whereas no differences in apoptosis were observed. VitD induced cell growth arrest and decreased proliferation in HULP cells (fold change = 0.74; P=.007). Moreover, VitD decreased Wnt-pathway expression in HULP cells at gene (activity score = −0.775; P<.001) and protein levels. However, VitD did not induce apoptosis expression.

      Conclusion

      Increased proliferation and Wnt/β-catenin pathway deregulation play a role in the development and growth of leiomyomas, whereas apoptosis appears not to contribute. VitD exerts an antiproliferative action on HULP cells through cell growth arrest and Wnt/β-catenin pathway inhibition, but not through apoptosis regulation, suggesting VitD as an effective therapy to stabilize leiomyoma size and prevent its growth.
      Inhibición de la proliferación de las células tumorales en miomas uterinos humanos mediante vitamina D a través de la ruta WNT/B-catenina

      Objetivo

      Evaluar el efecto de la vitamina D (Vit D) en miomas uterinos humanos mediante la inhibición de la ruta Wnt/b-catenina, inducción de la apoptosis y la detención del crecimiento celular.

      Diseño

      Estudio prospectivo para comparar miomas vs. tejidos del miometrio. Estudio de diseño pareado comparando células primarias de mioma uterino humano (HULP) tratadas con o sin VitD.

      Lugar de realización

      Hospital Universitario-Politécnico La Fe, Fundación IVI/INCLIVA, Universidad de Valencia, Valencia, España.

      Pacientes

      Miomas uterinos y tejido del miometrio fueron obtenidos de mujeres (edad 32-52 años) sin tratamiento hormonal.

      Intervención

      Las muestras fueron obtenidas de mujeres sometidas a cirugía debido a la patología de mioma uterino sintomático.

      Principales Variables de estudio

      Los tejidos del miometrio y mioma uterino fueron analizados por western blot (WB) para determinar la proliferación y las rutas de la Wnt/b-catenina y la apoptosis. Las células HULP fueron utilizadas para evaluar el efecto de la VitD sobre la proliferación celular (WB), el ciclo celular (citometría de flujo), genes de la Wnt/b-catenina y la apoptosis (arrays reacción en cadena de la polimerasa), las proteínas relacionadas con la Wnt (array proteínas) y la apoptosis (ensayo TUNEL).

      Resultados

      El tejido de mioma uterino, comparado con muestras iguales de miometrio, mostró una mayor proliferación (tasa de cambio 8.16;p=.0006) y la ruta Wnt/b-catenina alterada (tasa de cambio=5.5;p=<.0001), mientras que no se observó diferencias en la apoptosis. La VitD indujo la detención del crecimiento celular y disminuyó la proliferación en las células HULP (tasa de cambio=0.74; p=.007). Además, la VitD disminuyó la expresión de la ruta de la Wnt/b-catenina en células HULP tanto a nivel de los genes (Actividad_-0.775;P<.001) como de las proteínas. Sin embargo, la vitD no indujo la expresión de apoptosis.

      Conclusión

      El aumento de la proliferación y la desregulación de la ruta Wt/B-catenina tienen un papel importante en el desarrollo y crecimiento de los miomas; mientras que la apoptosis parece no contribuir. La VitD ejerce una acción antiproliferativa sobre las células HULP al detener el crecimiento celular e inhibir la ruta de la Wnt/b-catenina, pero no a través de la regulación de la apoptosis, sugiriendo que la VitD puede ser una terapia efectiva para estabilizar el tamaño del mioma y prevenir su crecimiento.

      Key Words

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