Yin Ping, Ono Masanori, Moravek Molly B, Coon John S, Navarro Antonia, Monsivais Diana, Dyson Matthew T, Druschitz Stacy A, Malpani Saurabh S, Serna Vanida A, Qiang Wenan, Chakravarti Debabrata, Kim J Julie, Bulun Serdar E
Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology (P.Y., M.O., M.B.M., J.S.C.V., A.N., D.M., M.T.D., S.A.D., S.S.M., W.Q., D.C., J.J.K., S.E.B.), Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611; and Department of Molecular and Cellular Biochemistry (V.A.S.), The Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.
J Clin Endocrinol Metab. 2015 Apr;100(4):E601-6. doi: 10.1210/jc.2014-2134. Epub 2015 Feb 6.
Uterine leiomyoma is the most common benign tumor in reproductive-age women. Using a dye-exclusion technique, we previously identified a side population of leiomyoma cells exhibiting stem cell characteristics. However, unless mixed with mature myometrial cells, these leiomyoma side population cells did not survive or grow well in vitro or in vivo.
The objective of this study was to identify cell surface markers to isolate leiomyoma stem/progenitor cells.
Real-time PCR screening was used to identify cell surface markers preferentially expressed in leiomyoma side population cells. In vitro colony-formation assay and in vivo tumor-regeneration assay were used to demonstrate functions of leiomyoma stem/progenitor cells.
We found significantly elevated CD49b and CD34 gene expression in side population cells compared with main population cells. Leiomyoma cells were sorted into three populations based on the expression of CD34 and CD49b: CD34(+)/CD49b(+), CD34(+)/CD49b(-), and CD34(-)/CD49b(-) cells, with the majority of the side population cells residing in the CD34(+)/CD49b(+) fraction. Of these populations, CD34(+)/CD49b(+) cells expressed the lowest levels of estrogen receptor-α, progesterone receptor, and α-smooth muscle actin, but the highest levels of KLF4, NANOG, SOX2, and OCT4, confirming their more undifferentiated status. The stemness of CD34(+)/CD49b(+) cells was also demonstrated by their strongest in vitro colony-formation capacity and in vivo tumor-regeneration ability.
CD34 and CD49b are cell surface markers that can be used to enrich a subpopulation of leiomyoma cells possessing stem/progenitor cell properties; this technique will accelerate efforts to develop new therapies for uterine leiomyoma.
子宫平滑肌瘤是育龄期女性最常见的良性肿瘤。我们之前使用染料排除技术鉴定出了具有干细胞特征的平滑肌瘤细胞侧群。然而，除非与成熟的子宫肌层细胞混合，这些平滑肌瘤侧群细胞在体外或体内均不能存活或良好生长。
本研究旨在鉴定细胞表面标志物以分离平滑肌瘤干/祖细胞。
采用实时PCR筛选来鉴定在平滑肌瘤侧群细胞中优先表达的细胞表面标志物。体外集落形成试验和体内肿瘤再生试验用于证明平滑肌瘤干/祖细胞的功能。
我们发现与主要群体细胞相比，侧群细胞中CD49b和CD34基因表达显著升高。根据CD34和CD49b的表达将平滑肌瘤细胞分为三个群体：CD34(+)/CD49b(+)、CD34(+)/CD49b(-)和CD34(-)/CD49b(-)细胞，大多数侧群细胞存在于CD34(+)/CD49b(+)部分。在这些群体中，CD34(+)/CD49b(+)细胞表达的雌激素受体-α、孕激素受体和α-平滑肌肌动蛋白水平最低，但KLF4、NANOG、SOX2和OCT4水平最高，证实了它们更未分化的状态。CD34(+)/CD49b(+)细胞的干性还通过其最强的体外集落形成能力和体内肿瘤再生能力得到证明。
CD34和CD49b是细胞表面标志物，可用于富集具有干/祖细胞特性的平滑肌瘤细胞亚群；该技术将加速开发子宫平滑肌瘤新疗法的努力。
