Department of Reproductive Endocrinology, Obstetrics & Gynecology Hospital, Fudan University, Shanghai 200011, China.
Hum Reprod. 2009 Nov;24(11):2879-89. doi: 10.1093/humrep/dep250. Epub 2009 Jul 24.
Galectin-3 (gal-3) is a beta-galactoside-binding protein which can be detected in endometrium. The study was designed to investigate synergism of gal-3 and integrinbeta3 in endometrial cell proliferation and adhesion in an in vitro model of endometrial receptivity.
The RL95-2 cell line was employed as an in vitro model for receptive endometrium. Cells transfected with gal-3 siRNA or treated with exogenous gal-3 were incubated with or without function-blocking integrinbeta1/3 antibody for evaluating synergism of gal-3 and integrins on cell proliferation and adhesion. Proliferation was measured by BrdU incorporation, and adhesion to fibronectin (FN) was determined by an adhesion assay. Integrin expression was analyzed by Flow Cytometry and western blots. Bewo spheroids were co-cultured with the RL95-2 monolayer to mimic the blastocyst-endometrial interaction, and colocalization of gal-3, integrinbeta3 and FN at the interface was observed by confocal microscopy.
The knock-down of gal-3 inhibited RL95-2 cell proliferation and adhesion. However, a reduction of proliferation and adhesion was also observed in presence of exogenous gal-3, and this was further reduced by a functional block to integrinbeta3. Moreover, gal-3 knock-down significantly increased integrinbeta3 expression, however, the colocalization of integrinbeta3 and FN was not increased. As expected, the colocalization of integrinbeta3 was decreased with the knock-down of gal-3.
This study has provided an in vitro model for the complex interactions between gal-3 and integrinbeta3 in the regulation of endometrial cell proliferation and adhesion.
半乳糖凝集素-3（gal-3）是一种可以在内膜中检测到的β-半乳糖苷结合蛋白。本研究旨在探讨 gal-3 与整合素β3 在子宫内膜容受性体外模型中的协同作用，以研究其对子宫内膜细胞增殖和黏附的影响。
采用 RL95-2 细胞系作为体外模型进行研究。用 gal-3 siRNA 转染细胞或用外源性 gal-3 处理细胞，然后与或不与功能阻断的整合素β1/3 抗体孵育，以评估 gal-3 和整合素在细胞增殖和黏附方面的协同作用。通过 BrdU 掺入法测量增殖，通过黏附试验测定黏附到纤维连接蛋白（FN）的能力。通过流式细胞术和 Western blot 分析整合素表达。将 Bewo 球体与 RL95-2 单层共培养，以模拟胚泡-子宫内膜相互作用，并通过共聚焦显微镜观察 gal-3、整合素β3 和 FN 在界面处的共定位。
gal-3 的敲低抑制了 RL95-2 细胞的增殖和黏附。然而，在外源性 gal-3 存在的情况下，增殖和黏附也减少了，而整合素β3 的功能阻断进一步减少了这种作用。此外，gal-3 的敲低显著增加了整合素β3 的表达，但整合素β3 和 FN 的共定位没有增加。正如预期的那样，gal-3 的敲低减少了整合素β3 的共定位。
本研究提供了一个体外模型，用于研究 gal-3 和整合素β3 在调节子宫内膜细胞增殖和黏附方面的复杂相互作用。
