Wegner C C, Cherington V, Clemens J W, Jacobs A L, Julian J, Surveyor G A, Bell E C, Carson D D
Department of Biochemistry and Molecular Biology, M.D. Anderson Cancer Center, Houston, Texas 77030, USA.
Endocrinology. 1996 Jan;137(1):175-84. doi: 10.1210/endo.137.1.8536610.
Uterine epithelial cells (UEC) isolated from 6-week-old CF-1 mice were immortalized using retroviral-mediated transfection of SV40 large T-antigen. One line, WEG-1, retained epithelial morphology and reacted with antibodies to cytokeratins 18, 19, laminin, fibronectin, and beta-catenin. In addition, WEG-1 cells displayed strong nuclear immunoreactivity to SV40 large T-antigen, confirming integration of the retrovirus vector and expression of this gene. WEG-1 cells were negative for nonepithelial markers such as desmin and factor 8. WEG-1 cells did not proliferate in serum-free medium; however, addition of 0.5% FBS supported proliferation to the same extent as 10% FBS. Addition of 50 ng/ml epidermal growth factor to medium containing 0.5% charcoal-stripped FBS restored proliferation comparable with 0.5% whole FBS. Epidermal growth factor or transforming growth factor-alpha (50 ng/ml), but not transforming growth factor-beta, leukemia-inhibiting factor, or fibroblast growth factor, induced the secretion of three proteins (M(r) approximately to 158K, 148K, and 36K). Comparison of protein secretions of WEG-1 cells and UEC showed shared as well as distinct bands. Like UEC, WEG-1 cells secreted PGF2a and PGE2 and expressed PG GH synthase-2. Unlike UEC, WEG-1 cells showed no apical/basal preference for either uptake of methionine or secretion of proteins. The absence of immunoreactive E-cadherin or zona occludens-1 was consistent with the absence of cell polarity in WEG-1 cells. Primary UEC, which polarize in vitro, do not support blastocyst attachment. WEG-1 cells, although not polarized in vitro, also exhibited delayed blastocyst attachment compared with nonuterine cell lines, suggesting that WEG-1 cells partially retained some aspects of UEC function relevant to embryo attachment. WEG-1 cells expressed messenger RNA for Muc-1, an UEC mucin suggested to have antiadhesive properties. Furthermore, WEG-1 cells did not display high affinity heparin binding sites, an activity associated with embryo attachment. WEG-1 cells may provide a model for studying various aspects of UEC function and murine embryo attachment.
从6周龄CF - 1小鼠分离出的子宫上皮细胞（UEC），通过逆转录病毒介导的SV40大T抗原转染实现永生化。其中一个细胞系WEG - 1保留了上皮形态，并与细胞角蛋白18、19、层粘连蛋白、纤连蛋白和β - 连环蛋白的抗体发生反应。此外，WEG - 1细胞对SV40大T抗原显示出强烈的核免疫反应性，证实了逆转录病毒载体的整合以及该基因的表达。WEG - 1细胞对诸如结蛋白和因子8等非上皮标记物呈阴性。WEG - 1细胞在无血清培养基中不增殖；然而，添加0.5%胎牛血清（FBS）支持其增殖，增殖程度与10% FBS相同。向含有0.5%活性炭处理的FBS的培养基中添加50 ng/ml表皮生长因子可恢复增殖，增殖程度与0.5%全FBS相当。表皮生长因子或转化生长因子 - α（50 ng/ml），而非转化生长因子 - β、白血病抑制因子或成纤维细胞生长因子，可诱导三种蛋白质（分子量约为158K、148K和36K）的分泌。WEG - 1细胞和UEC的蛋白质分泌比较显示有共同以及不同的条带。与UEC一样，WEG - 1细胞分泌前列腺素F2α（PGF2α）和前列腺素E2（PGE2）并表达前列腺素GH合酶 - 2。与UEC不同的是，WEG - 1细胞在蛋氨酸摄取或蛋白质分泌方面没有顶端/基底偏好。免疫反应性E - 钙黏蛋白或紧密连接蛋白 - 1的缺失与WEG - 1细胞中细胞极性的缺失一致。在体外极化的原代UEC不支持胚泡附着。WEG - 1细胞虽然在体外未极化，但与非子宫细胞系相比，胚泡附着也延迟，这表明WEG - 1细胞部分保留了与胚胎附着相关的UEC功能的某些方面。WEG - 1细胞表达黏蛋白1（Muc - 1）的信使核糖核酸，一种被认为具有抗黏附特性的UEC黏蛋白。此外，WEG - 1细胞未显示出高亲和力肝素结合位点，这是一种与胚胎附着相关的活性。WEG - 1细胞可能为研究UEC功能和小鼠胚胎附着的各个方面提供一个模型。
