Pavone Mary Ellen, Malpani Saurabh S, Dyson Matthew, Kim J Julie, Bulun Serdar E
Department of Obstetrics and Gynecology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
Department of Obstetrics and Gynecology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
Reprod Sci. 2016 Sep;23(9):1139-47. doi: 10.1177/1933719116632920. Epub 2016 Feb 25.
Fenretinide is a synthetic retinoid analogue that promotes apoptosis but has decreased toxicity when compared to other retinoids. We have previously shown that retinoic acid (RA) production in endometriotic tissue is decreased, resulting in reduced estrogen metabolism and apoptotic resistance. We hypothesize fenretinide may induce apoptosis in endometriotic cells and tissues, thereby reducing disease burden.
Primary endometriotic stromal cells were collected, isolated, cultured, and treated with fenretinide in doses from 0 to 20 µmol/L. Cell count, viability, and immunoblots were performed to examine apoptosis. Quantitative reverse transcription-polymerase chain reaction from endometriotic cells treated with fenretinide was used to examine expression of genes involved in RA signaling including stimulated by RA 6 (STRA6), cellular RA binding protein 2 (CRABP2), and fatty acid binding protein 5 (FABP5). Endometriotic tissue was xenografted subcutaneously into the flanks of mice which were treated with fenretinide for 2 weeks, after which the mice were killed and lesion volumes calculated. Statistical analysis was performed using t test and analysis of variance.
Treatment with fenretinide significantly decreased total cell count (doses 5-20 µL) and viability (doses 10-20 µmol/L). Fenretinide increased protein levels of the apoptotic marker poly (ADP ribose) polymerase (starting at 10 µmol/L) and decreased proliferation marker proliferating cell nuclear antigen (10 µmol/L, starting at 8-day treatment). Examination of genes involved in retinoid uptake and action showed that treatment induced STRA6 expression while expression of CRABP2 and FABP5 remained unchanged. Fenretinide also significantly decreased the endometriotic lesion xenograft volume.
Fenretinide increases STRA6 expression thereby potentially reversing the pathological loss of retinoid availability. Treatment with this compound induces apoptosis. In vivo treatments decrease lesion volume. Targeting the RA signaling pathway may be a promising novel treatment for women with endometriosis.
芬维A胺是一种合成类视黄醇类似物，可促进细胞凋亡，但与其他类视黄醇相比毒性降低。我们之前已经表明，子宫内膜异位症组织中视黄酸（RA）的产生减少，导致雌激素代谢降低和凋亡抵抗。我们假设芬维A胺可能诱导子宫内膜异位症细胞和组织凋亡，从而减轻疾病负担。
收集、分离、培养原发性子宫内膜异位症基质细胞，并用0至20 μmol/L剂量的芬维A胺处理。进行细胞计数、活力检测和免疫印迹以检测细胞凋亡。对用芬维A胺处理的子宫内膜异位症细胞进行定量逆转录-聚合酶链反应，以检测参与RA信号通路的基因表达，包括受RA刺激6（STRA6）、细胞RA结合蛋白2（CRABP2）和脂肪酸结合蛋白5（FABP5）。将子宫内膜异位症组织皮下移植到小鼠侧腹，用芬维A胺处理2周，之后处死小鼠并计算病变体积。使用t检验和方差分析进行统计分析。
芬维A胺处理显著降低了总细胞计数（5-20 μL剂量）和活力（10-20 μmol/L剂量）。芬维A胺增加了凋亡标志物聚（ADP核糖）聚合酶的蛋白水平（从10 μmol/L开始），并降低了增殖标志物增殖细胞核抗原（10 μmol/L，从第8天处理开始）。对视黄醇摄取和作用相关基因的检测表明，处理诱导了STRA6表达，而CRABP2和FABP5的表达保持不变。芬维A胺还显著降低了子宫内膜异位症病变移植瘤体积。
芬维A胺增加STRA6表达，从而可能逆转类视黄醇可用性的病理损失。用该化合物处理可诱导细胞凋亡。体内处理可减小病变体积。靶向RA信号通路可能是子宫内膜异位症女性一种有前景的新治疗方法。
