Over Expression of CD44V8—10 in Human Bladder Cancer Cells Decreases Their Interaction With Hyaluronic Acid and Potentiates Their Malignant Progression

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Over Expression of CD44V8—10 in Human Bladder Cancer Cells Decreases Their Interaction With Hyaluronic Acid and Potentiates Their Malignant Progression MOTOTSUGU MURAMAKI, HIDEAKI MIYAKE, SADAO KAMIDONO, ISAO HARA The Journal of Urology Volume 171, Issue 1, Pages 426-430 (January 2004) DOI: 10.1097/01.ju.0000093446.54115.b6 Copyright © 2004 American Urological Association, Inc. Terms and Conditions

Fig. 1 A, RT-PCR amplification of cDNA of UM-UC-3/P, that is parental cell line of UM-UC-3, UM-UC-3/C, that is vector only transfected cell line, and 2 clones of CD44V8–10 transfectant UM-UC-3/v8–10 No. 1 and No. 2. B, Western blot analysis of cell lysates derived from UM-UC-3/P, UM-UC-3/C, and UM-UC-3/v8–10 No. 1 and No. 2. The Journal of Urology 2004 171, 426-430DOI: (10.1097/01.ju.0000093446.54115.b6) Copyright © 2004 American Urological Association, Inc. Terms and Conditions

Fig. 2 In vitro proliferation of UM-UC-3 sublines after 1 × 103 cells per line were seeded in 6-well plates. Number of cells per cell line was counted daily in triplicate. Bars represent SD. The Journal of Urology 2004 171, 426-430DOI: (10.1097/01.ju.0000093446.54115.b6) Copyright © 2004 American Urological Association, Inc. Terms and Conditions

Fig. 3 Adhesion assay of UM-UC-3 sublines examined for adhesion to various concentrations of extracellular matrix proteins. A, HA. B, 1 μg/ml fibronectin, 25 μg/ml laminin, 1 mg/ml type IV collagen and 25 μg/ml vitronectin. Columns and bars represent mean ± SD. Asterisk indicates Student’s t test p <0.01 vs UM-UC-3/v8–10 No. 1 and No. 2. The Journal of Urology 2004 171, 426-430DOI: (10.1097/01.ju.0000093446.54115.b6) Copyright © 2004 American Urological Association, Inc. Terms and Conditions

Fig. 4 Chemotactic effect of HA on UM-UC-3 sublines. Number of migrated cells per cell line was compared by migration assay using HA as chemoattractant. Columns and bars represent mean ± SD. Asterisk indicates Student’s t test p <0.01 vs UM-UC-3/v8–10 No. 1 and No. 2. The Journal of Urology 2004 171, 426-430DOI: (10.1097/01.ju.0000093446.54115.b6) Copyright © 2004 American Urological Association, Inc. Terms and Conditions

Fig. 5 Tumor growth of UM-UC-3 sublines in athymic nude mice subcutaneously administrated each cell line in right flank on day 0. Tumor volume was measured twice weekly and calculated using formula, length × width × depth × 0.5236. Bars represent SD. Asterisk indicates Student’s t test p <0.05 vs UM-UC-3/P and UM-UC-3/C. The Journal of Urology 2004 171, 426-430DOI: (10.1097/01.ju.0000093446.54115.b6) Copyright © 2004 American Urological Association, Inc. Terms and Conditions