Intracellular concentration and structural states of actin in human bladder carcinomas: New indices for malignancy.

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In order to determine whether the analysis of actin, a cytoskeletal protein, can be useful to predict the growth pattern and invasiveness of bladder carcinoma, intracellular concentration and structural states of actin were examined in five established cell lines derived from human bladder carcinomas of different pathological grades (KU7: grade 1, KU1: grade 2, T24: grade 3, MGH-UI, NBT-2: grade 4), in surgically obtained cancer cells from patients with bladder carcinoma, and in cells from normal transitional epithelium of the bladder.<br>Distribution of actin in the cytoplasm of above mentioned cells was successfully observed by immunofluorescent technique utilizing anti-actin monoclonal antibody. Concentration of actin in the cells was determined by DNAase I inhibition assay and by a densitometrical measurement after electrophoretic separation of the cell component. Distribution of fibronectin (FN) and cell surface receptor of concanavarin A (con A) were also assessed by immunofluorescent technique.<br>The results revealed that aggregated fluorescent bundles, i.e. stress fibers (SFs), were clearly seen in KU7 and T24 cells, and surgically obtained cells from patients with low grade carcinoma, whereas only diffusely distributed fluorescence was observed in NBT-2 cells and surgical specimens from high grade carcinoma.<br>Concentrations of actin in cultured cell lines, KU7, KU1, T24, MGH-UI and NBT-2, were 38.1±4.0, 37.0±1.0, 34.5±1.0, 32.1±0.5, 25.2±0.7 (mean±SD μg/mg protein), respectively, while those of surgical specimens of normal bladder epithelial cells, non invasive bladder carcinoma cells, and invasive bladder carcinoma cells were 65.0±3.4, 52.8±5.2, 35.4±4.4 (mean±SD μg/mg protein), respectively. Thus, high grade bladder carcinoma cells showed lower concentrations of actin compared with those of low grade bladder carcinoma cells. Distribution of cell surface FN and rate of capping of con A receptors were well correlated with the intracellular concentration of actin and the configuration of SFs.<br>These results indicate that determination of intracellular concentration of actin provides a new modality to characterize the biological behaviors, i.e. the growth pattern and invasivness, of bladder carcinoma.

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