Kufaishi Hala, Alarab May, Drutz Harold, Lye Stephen, Shynlova Oksana
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Mount Sinai Hospital, Toronto, Ontario, Canada Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada.
Reprod Sci. 2016 Jul;23(7):931-43. doi: 10.1177/1933719115625840. Epub 2016 Jan 13.
This study tested a hypothesis that primary human vaginal cells derived from tissue of premenopausal women with severe pelvic organ prolapse (POP-HVCs) would display differential functional characteristics as compared to vaginal cells derived from asymptomatic women with normal pelvic floor support (control-HVCs).
Vaginal tissue biopsies were collected from premenopausal patients with POP (n = 8) and asymptomatic controls (n = 7) during vaginal hysterectomy or repair. Primary vaginal cells were isolated by enzymatic digestion and characterized by immunocytochemistry. Cell attachment and proliferation on different matrices (collagen I, collagen II, collagen IV, fibronectin, laminin, tenascin, and vitronectin) were compared between POP-HVCs and control-HVCs. RNA was extracted, and the expression of 84 genes was screened using Human Extracellular Matrix and Adhesion Molecules RT(2) Profiler PCR array. The expression of selected genes was verified by quantitative reverse transcription-polymerase chain reaction.
(1) Control-HVCs attached to collagen IV more efficiently than POP-HVCs; (2) control-HVCs and POP-HVCs show a similar proliferation rate when plated on proNectin and collagen I; (3) when seeded on collagen I, resting POP-HVCs expressed significantly (P < .05) increased transcript levels of collagen VII, multiple matrix metalloproteinases (MMP3, MMP7, MMP10, MMP12, MMP13, and MMP14), integrins (ITGA1, ITGA4, ITGA6, ITGA8, ITGB1, ITGB2, and ITGB3), and cell adhesion molecules as compared to control-HVCs. Collagen XV and tissue inhibitors of MMPs (TIMP1 and TIMP2) as well as genes involved in the biogenesis and maturation of collagen and elastin fibers (LOX, LOXL1-LOXL3, BMP1, and ADAMTS2) were significantly downregulated in POP-HVCs versus control-HVCs (P < .05).
Resting primary POP-HVCs in vitro show altered cellular characteristics as compared to control-HVCs, which may influence their dynamic responses to external mechanical or hormonal stimuli.
本研究检验了一个假设，即与来自盆底支持正常的无症状女性的阴道细胞（对照-人阴道细胞）相比，源自严重盆腔器官脱垂绝经前女性组织的原代人阴道细胞（盆腔器官脱垂-人阴道细胞）会表现出不同的功能特征。
在阴道子宫切除术或修复过程中，从患有盆腔器官脱垂的绝经前患者（n = 8）和无症状对照者（n = 7）采集阴道组织活检样本。通过酶消化分离原代阴道细胞，并采用免疫细胞化学法进行鉴定。比较盆腔器官脱垂-人阴道细胞和对照-人阴道细胞在不同基质（I型胶原、II型胶原、IV型胶原、纤连蛋白、层粘连蛋白、腱生蛋白和玻连蛋白）上的细胞黏附及增殖情况。提取RNA，使用人细胞外基质和黏附分子RT(2) Profiler PCR芯片筛选84个基因的表达。通过定量逆转录-聚合酶链反应验证所选基因的表达。
（1）对照-人阴道细胞比盆腔器官脱垂-人阴道细胞更有效地黏附于IV型胶原；（2）当接种于亲黏蛋白和I型胶原上时，对照-人阴道细胞和盆腔器官脱垂-人阴道细胞显示出相似的增殖率；（3）当接种于I型胶原上时，静息状态的盆腔器官脱垂-人阴道细胞与对照-人阴道细胞相比，VII型胶原、多种基质金属蛋白酶（MMP3、MMP7、MMP10、MMP12、MMP13和MMP14）、整合素（ITGA1、ITGA4、ITGA6、ITGA8ITGB1、ITGB2和ITGB3）以及细胞黏附分子的转录水平显著升高（P < 0.05）。与对照-人阴道细胞相比，盆腔器官脱垂-人阴道细胞中XV型胶原和基质金属蛋白酶组织抑制剂（TIMP1和TIMP2）以及参与胶原和弹性纤维生物合成与成熟的基因（LOX、LOXL1-LOXL3、BMP1和ADAMTS2）显著下调（P < 0.05）。
与对照-人阴道细胞相比，体外静息状态的原代盆腔器官脱垂-人阴道细胞显示出细胞特征改变，这可能影响其对外部机械或激素刺激的动态反应。
