Department of Gynecology and Obstetrics, Division of Reproductive Endocrinology and Infertility, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Department of Gynecology and Obstetrics, Division of Reproductive Sciences and Women's Health Research, Johns Hopkins Medicine, Baltimore, Maryland.
F S Sci. 2021 Aug;2(3):303-314. doi: 10.1016/j.xfss.2021.04.002. Epub 2021 Apr 13.
To determine if A-kinase anchoring protein 13 (AKAP13) interacts with the vitamin D receptor (VDR) to alter vitamin D-dependent signaling in fibroid cells. Uterine leiomyomas (fibroids) are characterized by a fibrotic extracellular matrix and are associated with vitamin D deficiency. Treatment with vitamin D (1,25-dihydroxyvitamin D) reduces fibroid growth and extracellular matrix gene expression. A-kinase anchoring protein 13 is overexpressed in fibroids and interacts with nuclear hormone receptors, but it is not known whether AKAP13 may interact with the VDR to affect vitamin D signaling in fibroids.
Laboratory studies.
Translational science laboratory.
INTERVENTION(S): Human immortalized fibroid or myometrial cells were treated with 1,25-hydroxyvitamin D (1,25(OH)D) and transfected using expression constructs for AKAP13 or AKAP13 mutants, RhoQL, C3 transferase, or small interfering ribonucleic acids (RNAs).
MAIN OUTCOME MEASURE(S): Messenger ribonucleic acid (mRNA) levels of AKAP13, fibromodulin, and versican as measured by quantitative real-time polymerase chain reaction. Glutathione S-transferase-binding assays. Vitamin D-dependent gene activation as measured by luciferase assays.
RESULT(S): 1,25(OH)D resulted in a significant reduction in mRNA levels encoding AKAP13, versican, and fibromodulin. Small interfering RNA silencing of AKAP13 decreased both fibromodulin and versican mRNA levels. Glutathione S-transferase-binding assays revealed that AKAP13 bound to the VDR through its nuclear receptor interacting region. Cotransfection of AKAP13 and VDR significantly reduced vitamin D-dependent gene activation. RhoA pathway inhibition partially relieved repression of vitamin D-dependent gene activation by AKAP13.
CONCLUSION(S): These data suggest that AKAP13 inhibited the vitamin D receptor activation by a mechanism that required, at least in part, RhoA activation.
确定激酶锚定蛋白 13（AKAP13）是否与维生素 D 受体（VDR）相互作用，从而改变纤维瘤细胞中维生素 D 依赖性信号转导。子宫平滑肌瘤（纤维瘤）的特征是细胞外基质纤维化，并与维生素 D 缺乏有关。维生素 D（1,25-二羟维生素 D）治疗可减少纤维瘤生长和细胞外基质基因表达。AKAP13 在纤维瘤中过度表达，并与核激素受体相互作用，但尚不清楚 AKAP13 是否可能与 VDR 相互作用，从而影响纤维瘤中的维生素 D 信号转导。
实验室研究。
转化科学实验室。
用 1,25-羟基维生素 D（1,25（OH）D）处理人永生化纤维瘤或子宫平滑肌细胞，并使用 AKAP13 或 AKAP13 突变体、RhoQL、C3 转移酶或小干扰核糖核酸（RNA）的表达构建体转染。
定量实时聚合酶链反应测定信使核糖核酸（mRNA）水平的 AKAP13、纤维调节素和 versican。谷胱甘肽 S-转移酶结合测定。通过荧光素酶测定维生素 D 依赖性基因激活。
1,25（OH）D 可显著降低编码 AKAP13、versican 和纤维调节素的 mRNA 水平。AKAP13 的小干扰 RNA 沉默降低了纤维调节素和 versican 的 mRNA 水平。谷胱甘肽 S-转移酶结合测定表明 AKAP13 通过其核受体相互作用区与 VDR 结合。AKAP13 和 VDR 的共转染显著降低了维生素 D 依赖性基因激活。RhoA 途径抑制部分缓解了 AKAP13 对维生素 D 依赖性基因激活的抑制作用。
这些数据表明，AKAP13 通过至少部分需要 RhoA 激活的机制抑制维生素 D 受体的激活。
