Abstract
Rabbit bladder epithelium, grown on collagen gels and exposed to the chemical carcinogen benzo[a]pyrene, produced nontumorigenic altered foci as well as tumorigenic epithelial cell lines during 120-180 d in culture. Immunofluorescence studies revealed extensive keratin filaments in both primary epithelial cells and benzo[a]pyrene-induced altered epithelial foci but showed no detectable vimentin filaments. The absence of vimentin expression in these cells was confirmed by two- dimensional gel electrophoresis. In contrast, immunofluorescence staining of the cloned benzo[a]pyrene-transformed rabbit bladder epithelial cell line, RBC-1, revealed a reduction in filamentous keratin concomitant with the expression of vimentin filaments. The epithelial nature of this cell line was established by the observation that cells injected into nude mice formed well-differentiated adenocarcinomas. Frozen sections of such tumors showed strong staining with antikeratins antibodies, but no detectable staining with antivimentin antibodies. These results demonstrated a differential expression of intermediate filament type in cells at different stages of neoplastic progression and in cells maintained in different growth environments. It is apparent that the expression of intermediate filaments throughout neoplastic progression is best studied by use of an in vivo model system in parallel with culture studies.
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- Bennett G. S., Fellini S. A., Croop J. M., Otto J. J., Bryan J., Holtzer H. Differences among 100-A filamentilament subunits from different cell types. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4364–4368. doi: 10.1073/pnas.75.9.4364. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett G. S., Fellini S. A., Holtzer H. Immunofluorescent visualization of 100 A filaments in different cultured chick embryo cell types. Differentiation. 1978;12(2):71–82. doi: 10.1111/j.1432-0436.1979.tb00992.x. [DOI] [PubMed] [Google Scholar]
- Bignami A., Dahl D. Astrocyte-specific protein and neuroglial differentiation. An immunofluorescence study with antibodies to the glial fibrillary acidic protein. J Comp Neurol. 1974 Jan 1;153(1):27–38. doi: 10.1002/cne.901530104. [DOI] [PubMed] [Google Scholar]
- Blose S. H. Ten-nanometer filaments and mitosis: maintenance of structural continuity in dividing endothelial cells. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3372–3376. doi: 10.1073/pnas.76.7.3372. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cervera M., Dreyfuss G., Penman S. Messenger RNA is translated when associated with the cytoskeletal framework in normal and VSV-infected HeLa cells. Cell. 1981 Jan;23(1):113–120. doi: 10.1016/0092-8674(81)90276-2. [DOI] [PubMed] [Google Scholar]
- Doran T. I., Vidrich A., Sun T. T. Intrinsic and extrinsic regulation of the differentiation of skin, corneal and esophageal epithelial cells. Cell. 1980 Nov;22(1 Pt 1):17–25. doi: 10.1016/0092-8674(80)90150-6. [DOI] [PubMed] [Google Scholar]
- Fellini S. A., Bennett G. S., Holtzer H. Selective binding of antibody against gizzard 10-nm filaments to different cell types in myogenic cultures. Am J Anat. 1978 Nov;153(3):451–457. doi: 10.1002/aja.1001530308. [DOI] [PubMed] [Google Scholar]
- Fellini S. A., Bennett G. S., Toyama Y., Holtzer H. Biochemical and immunological heterogeneity of 100 A filament subunits from different chick cell types. Differentiation. 1978;12(2):59–69. doi: 10.1111/j.1432-0436.1979.tb00991.x. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Appelhans B., Schmid E., Freudenstein C., Osborn M., Weber K. Identification and characterization of epithelial cells in mammalian tissues by immunofluorescence microscopy using antibodies to prekeratin. Differentiation. 1979;15(1):7–25. doi: 10.1111/j.1432-0436.1979.tb01030.x. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Grund C., Osborn M., Weber K. The intermediate-sized filaments in rat kangaroo PtK2 cells. I. Morphology in situ. Cytobiologie. 1978 Aug;17(2):365–391. [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Osborn M., Weber K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5034–5038. doi: 10.1073/pnas.75.10.5034. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
- Garrels J. I. Two dimensional gel electrophoresis and computer analysis of proteins synthesized by clonal cell lines. J Biol Chem. 1979 Aug 25;254(16):7961–7977. [PubMed] [Google Scholar]
- Gordon W. E., 3rd, Bushnell A., Burridge K. Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. Cell. 1978 Feb;13(2):249–261. doi: 10.1016/0092-8674(78)90194-0. [DOI] [PubMed] [Google Scholar]
- Hynes R. O., Destree A. T. 10 nm filaments in normal and transformed cells. Cell. 1978 Jan;13(1):151–163. doi: 10.1016/0092-8674(78)90146-0. [DOI] [PubMed] [Google Scholar]
- Knowles M. A., Franks L. M. Stages in neoplastic transformation of adult epithelial cells by 7,12-dimethylbenz(a)anthracene in vitro. Cancer Res. 1977 Nov;37(11):3917–3924. [PubMed] [Google Scholar]
- Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
- Lazarides E. The distribution of desmin (100 A) filaments in primary cultures of embryonic chick cardiac cells. Exp Cell Res. 1978 Mar 15;112(2):265–273. doi: 10.1016/0014-4827(78)90209-4. [DOI] [PubMed] [Google Scholar]
- Liem R. K., Yen S. H., Salomon G. D., Shelanski M. L. Intermediate filaments in nervous tissues. J Cell Biol. 1978 Dec;79(3):637–645. doi: 10.1083/jcb.79.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marshall C. J., Franks L. M., Carbonell A. W. Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas. J Natl Cancer Inst. 1977 Jun;58(6):1743–1751. doi: 10.1093/jnci/58.6.1743. [DOI] [PubMed] [Google Scholar]
- Michalopoulos G., Pitot H. C. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp Cell Res. 1975 Aug;94(1):70–78. doi: 10.1016/0014-4827(75)90532-7. [DOI] [PubMed] [Google Scholar]
- Osborn M., Weber K. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane. Proc Natl Acad Sci U S A. 1976 Mar;73(3):867–871. doi: 10.1073/pnas.73.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlaepfer W. W., Lynch R. G. Immunofluorescence studies of neurofilaments in the rat and human peripheral and central nervous system. J Cell Biol. 1977 Jul;74(1):241–250. doi: 10.1083/jcb.74.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmid E., Tapscott S., Bennett G. S., Croop J., Fellini S. A., Holtzer H., Franke W. W. Differential location of different types of intermediate-sized filaments in various tissues of the chicken embryo. Differentiation. 1979;15(1):27–40. doi: 10.1111/j.1432-0436.1979.tb01031.x. [DOI] [PubMed] [Google Scholar]
- Starger J. M., Brown W. E., Goldman A. E., Goldman R. D. Biochemical and immunological analysis of rapidly purified 10-nm filaments from baby hamster kidney (BHK-21) cells. J Cell Biol. 1978 Jul;78(1):93–109. doi: 10.1083/jcb.78.1.93. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Summerhayes I. C., Franks L. M. Effects of donor age on neoplastic transformation of adult mouse bladder epithelium in vitro. J Natl Cancer Inst. 1979 Apr;62(4):1017–1023. [PubMed] [Google Scholar]
- Sun T. T., Doran T. I., Vidrich A. The use of antikeratin antibodies for the identification of cultured epithelial cells. Birth Defects Orig Artic Ser. 1980;16(2):183–196. [PubMed] [Google Scholar]
- Sun T. T., Green H. Immunofluorescent staining of keratin fibers in cultured cells. Cell. 1978 Jul;14(3):469–476. doi: 10.1016/0092-8674(78)90233-7. [DOI] [PubMed] [Google Scholar]
- Sun T. T., Green H. Keratin filaments of cultured human epidermal cells. Formation of intermolecular disulfide bonds during terminal differentiation. J Biol Chem. 1978 Mar 25;253(6):2053–2060. [PubMed] [Google Scholar]
- Sun T. T., Shih C., Green H. Keratin cytoskeletons in epithelial cells of internal organs. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2813–2817. doi: 10.1073/pnas.76.6.2813. [DOI] [PMC free article] [PubMed] [Google Scholar]