Di Cicco L M, Mansbridge J N, Morhenn V B
Papanicolaou Comprehensive Cancer Center, University of Miami.
In Vitro Cell Dev Biol. 1987 Dec;23(12):805-14. doi: 10.1007/BF02620958.
A murine monoclonal antibody, VM-1, which binds to basal cells of normal human epidermis, reduces the ability of human squamous cell carcinoma cells (SCL-1) derived from the skin to attach and spread on collagen by about 50% and causes cell rounding. Similar effects have been previously shown using normal human keratinocytes. The attachment of cell lines derived from human lung squamous cell carcinomas (SW1271 and SW900), melanoma A375, glioblastoma 126, and fibrosarcoma HT1080 is also inhibited by this antibody. VM-1 antibody does not bind to normal human fibroblasts, benign nevus cells, or the human B-cell-derived line 8866. VM-1 antibody inhibits the growth of SCL-1 cells in vitro as measured by cell numbers and [3H]thymidine ([3H]TdR) incorporation. It is not cytolytic in the presence of complement as measured by 51Cr release. Repeated treatment of SCL-1 cells with VM-1 antibody significantly reduces the proportion of SCL-1 cells that attach to collagen. In addition, after treatment of SCL-1 cells with VM-1 antibody, several proteins can no longer be demonstrated by gel electrophoresis of the cell-free supernatant. The VM-1 antibody effect on attachment and spreading is partially reversed by pretreatment of the collagen surface with laminin and fibronectin, but not with the carbohydrates chondroitin-6-sulfate or hyaluronic acid or with the protein lysozyme. By fluorescence staining, the antigen recognized by VM-1 antibody is membrane-bound and Triton X-100 extractable. The VM-1 antigen is excluded from Bio-Sil TSK-400 and sediments at about 10.5 S. It has a covalent molecular weight on the order of 10(6). Proteinase K digestion produces VM-1 antibody reactive fragments, assumed to be polysaccharides, with a polydisperse molecular weight distribution in the range 5000 to 30,000. The VM-1 antigen is partially lost from solution on boiling and is no longer detectable in the aqueous or organic phase after chloroform-methanol extraction. The properties of the VM-1 antigen are consistent with those of a proteoglycan involved in attachment and spreading of keratinocytes and certain tumor cells on collagen.
一种鼠单克隆抗体VM - 1，可与正常人表皮的基底细胞结合，使源自皮肤的人鳞状细胞癌细胞（SCL - 1）在胶原蛋白上附着和扩散的能力降低约50%，并导致细胞变圆。先前使用正常人角质形成细胞也显示出类似的效果。源自人肺鳞状细胞癌（SW1271和SW900）、黑色素瘤A375、胶质母细胞瘤126和纤维肉瘤HT1080的细胞系的附着也受到该抗体的抑制。VM - 1抗体不与正常人成纤维细胞、良性痣细胞或人B细胞衍生系8866结合。VM - 1抗体在体外抑制SCL - 1细胞的生长，通过细胞数量和[³H]胸腺嘧啶核苷（[³H]TdR）掺入来衡量。通过⁵¹Cr释放测量，在补体存在下它没有细胞毒性。用VM - 1抗体反复处理SCL - 1细胞可显著降低附着于胶原蛋白的SCL - 1细胞的比例。此外，用VM - 1抗体处理SCL - 1细胞后，通过对无细胞上清液进行凝胶电泳，几种蛋白质不再显示。VM - 1抗体对附着和扩散的作用可通过用层粘连蛋白和纤连蛋白预处理胶原蛋白表面而部分逆转，但用硫酸软骨素 - 6 - 硫酸盐或透明质酸等碳水化合物或溶菌酶等蛋白质预处理则不能。通过荧光染色，VM - 1抗体识别的抗原是膜结合的且可被Triton X - 100提取。VM - 1抗原被Bio - Sil TSK - 400排除，并在约10.5 S处沉降。其共价分子量约为10⁶。蛋白酶K消化产生VM - 1抗体反应性片段，推测为多糖，分子量分布多分散，范围在5000至30000之间。VM - 1抗原在煮沸时部分从溶液中丢失，在氯仿 - 甲醇提取后在水相或有机相中不再可检测到。VM - 1抗原的特性与参与角质形成细胞和某些肿瘤细胞在胶原蛋白上附着和扩散的蛋白聚糖的特性一致。
