Casey Rachael C, Koch Kimberly A, Oegema Theodore R, Skubitz Keith M, Pambuccian Stefan E, Grindle Suzanne M, Skubitz Amy P N
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Clin Exp Metastasis. 2003;20(4):343-56. doi: 10.1023/a:1024009131191.
Ovarian carcinoma is the leading cause of gynecological cancer deaths in the United States. Secondary tumor growths form by tumor cell invasion through the mesothelial lining of the peritoneal cavity and peritoneal organs. To study this interaction, we developed a dye-based in vitro model system in which mesothelial cells were grown as confluent monolayers, permeabilized, and then co-cultured with ovarian carcinoma cells for up to seven days. The mesothelial cells were then stained with trypan blue dye, which enabled the visualization of ovarian carcinoma cell invasion through the monolayers of mesothelial cells. Ovarian carcinoma cell invasion was inhibited for up to 7 days by the addition of GRGDSP peptides, a blocking monoclonal antibody against the beta1 integrin subunit, or blocking monoclonal antibodies against matrix metalloproteinases 2 and 9. Cell invasion was also inhibited by hyaluronan and GM6001, a chemical inhibitor of matrix metalloproteinases. Differential gene expression of matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases, and disintegrins were observed in primary ovarian carcinoma tumors and secondary metastases, compared to normal ovaries. Taken together, these results suggest that complex interactions between integrins, disintegrins, matrix metalloproteinases, and tissue inhibitors of matrix metalloproteinases may mediate ovarian carcinoma cell invasion, and that the dye-based assay described herein is a suitable model system for its study.
卵巢癌是美国妇科癌症死亡的主要原因。继发性肿瘤生长是由肿瘤细胞通过腹膜腔和腹膜器官的间皮衬里侵入而形成的。为了研究这种相互作用，我们开发了一种基于染料的体外模型系统，其中间皮细胞生长为汇合的单层，使其通透化，然后与卵巢癌细胞共培养长达7天。然后用台盼蓝染料对间皮细胞进行染色，这使得能够观察到卵巢癌细胞穿过间皮细胞单层的侵袭情况。通过添加GRGDSP肽、抗β1整合素亚基的阻断单克隆抗体或抗基质金属蛋白酶2和9的阻断单克隆抗体，卵巢癌细胞的侵袭被抑制长达7天。透明质酸和GM6001（一种基质金属蛋白酶的化学抑制剂）也抑制了细胞侵袭。与正常卵巢相比，在原发性卵巢癌肿瘤和继发性转移灶中观察到基质金属蛋白酶、基质金属蛋白酶组织抑制剂和去整合素的差异基因表达。综上所述，这些结果表明整合素、去整合素、基质金属蛋白酶和基质金属蛋白酶组织抑制剂之间的复杂相互作用可能介导卵巢癌细胞的侵袭，并且本文所述的基于染料的检测方法是研究其的合适模型系统。
