Development of screening assay for cancer invasion inhibitory factors using human cell lines and its application to tea catechins
1998
Yamamoto, M.M. (National Research Inst. of Vegetables, Ornamental Plants and Tea, Ano, Mie (Japan)) | Kawahara, H. | Osada, K. | Yamaguchi, Y. | Tsuji, K.
In medical treatments of cancer, it is important to inhibit cancer cell metastasis. In the process of cancer cell metastasis, interactions between metastatic cancer cells and vein endothelial cells, such as cancer cell adhesion and invasion to endothelial cells, are important. Recently, an in vitro assay system to screen the inhibitory factors on cancer metastasis has been developed using animal cultured cells. In the present study, we selected a human cancer cell line for in vitro invasion assay in the combined human cells culture system as follows: Human umbilical vein endothelial cells (HUV-EC-C) were cultured on a gelatin pre-coated insert membrane with 8 mu-m pores. Five florescence labeled human cancer cell lines (MKN-45(gastric cancer), MCF-7(breast cancer), A549(lung cancer), T-24(bladder cancer) and HT-1080(fibrosarcoma)) were added to HUV-EC-C monolayers. After incubation for 5 hours invasive cancer cells on each lower surface of insert membranes were collected and the cell number was determined by measurement of fluorescent intensity. Among five human cancer cell lines, only human fibrosarcoma HT-1080 showed invasiveness as well as adherence to HUV-EC-C/basement membrane. The application of this in vitro assay system to screening for cancer invasion inhibitory factors showed that tea ester-type catechins with gallate-group had an invasion inhibitory activity in in vivo assay as reported previously. Especially, (-)-epicatechin gallate showed an strong invasion inhibitory activity under the concentration showed cell toxicity
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