Cytoplasmic microtubules in two different mouse melanoma cell lines: a qualitative and quantitative analysis using confocal laser scanning microscopy and computer-assisted image analysis
The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma ce...
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| Published in | Journal of cutaneous pathology Vol. 24; no. 6; pp. 350 - 355 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford, UK
Blackwell Publishing Ltd
01.07.1997
Blackwell |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0303-6987 1600-0560 |
| DOI | 10.1111/j.1600-0560.1997.tb00803.x |
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| Abstract | The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma cell lines (K1735‐M2: high metastatic clone; K1735‐cl16: low metastatic clone) were determined quantitatively using an indirect immunofluorescence technique, confocal laser scanning microscopy (CLSM)and computer‐assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule‐inhibitor nocodazole were investigated. Quantitative analysis showed a significant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the high metastatic cells. After treatment with nocodazole the parameters TEXTURE and DENSITY were reduced, suggesting a decrease of assembled tubulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer‐assisted image analysis provides a new method to examine quantitative variations of the cytoskeleton possibly related to cell function. |
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| AbstractList | The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma cell lines (K1735‐M2: high metastatic clone; K1735‐cl16: low metastatic clone) were determined quantitatively using an indirect immunofluorescence technique, confocal laser scanning microscopy (CLSM)and computer‐assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule‐inhibitor nocodazole were investigated. Quantitative analysis showed a significant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the high metastatic cells. After treatment with nocodazole the parameters TEXTURE and DENSITY were reduced, suggesting a decrease of assembled tubulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer‐assisted image analysis provides a new method to examine quantitative variations of the cytoskeleton possibly related to cell function. The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma cell lines (K1735-M2: high metastatic clone; K1735-c116: low metastatic clone) were determined quantitatively using an indirect immunofluorescence technique, confocal laser scanning microscopy (CLSM) and computer-assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule-inhibitor nocodazole were investigated. Quantitative analysis showed a significant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the high metastatic cells. After treatment with nocodazole the parameters TEXTURE and DENSITY were reduced, suggesting a decrease of assembled tubulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer-assisted image analysis provides a new method to examine quantitative variations of the cytoskeleton possibly related to cell function.The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma cell lines (K1735-M2: high metastatic clone; K1735-c116: low metastatic clone) were determined quantitatively using an indirect immunofluorescence technique, confocal laser scanning microscopy (CLSM) and computer-assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule-inhibitor nocodazole were investigated. Quantitative analysis showed a significant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the high metastatic cells. After treatment with nocodazole the parameters TEXTURE and DENSITY were reduced, suggesting a decrease of assembled tubulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer-assisted image analysis provides a new method to examine quantitative variations of the cytoskeleton possibly related to cell function. The microtubular system as one part of the cellular cytoskeleton is not only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastases. In the present study, the amount and distribution of tubulin in two murine melanoma cell lines (K1735-M2: high metastatic clone; K1735-c116: low metastatic clone) were determined quantitatively using an indirect immunofluorescence technique, confocal laser scanning microscopy (CLSM) and computer-assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule-inhibitor nocodazole were investigated. Quantitative analysis showed a significant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the high metastatic cells. After treatment with nocodazole the parameters TEXTURE and DENSITY were reduced, suggesting a decrease of assembled tubulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer-assisted image analysis provides a new method to examine quantitative variations of the cytoskeleton possibly related to cell function. |
| Author | Helige, Christine Hofmann-Wellenhof, Rainer Smolle, Josef Kerl, Helmut Fink-Puches, Regina |
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| Cites_doi | 10.1016/0014-4827(87)90291-6 10.1007/BF00119079 10.1016/S1040-8428(85)80026-3 10.1172/JCI113384 10.1038/321605a0 10.1002/9780470719978.ch6 10.1093/jnci/63.3.541 10.1242/jcs.87.4.535 10.1242/jcs.104.2.339 10.1083/jcb.105.1.41 10.1016/0277-5379(82)90064-5 10.1097/00000372-199002000-00003 10.1073/pnas.72.3.994 10.1111/j.1600-0560.1995.tb00747.x 10.1073/pnas.72.12.4981 10.1007/BF00046338 10.1016/0959-8049(93)90589-8 10.1016/0190-9622(93)70158-P |
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| Keywords | Cell culture Scanning electron microscopy Rodentia Melanoma Exploration Experimental study Pathology Vertebrata Image analysis Mammalia Mouse Animal Laser Tumor Cytoskeleton Qualitative analysis Microtubule Computer aid Quantitative analysis |
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| References | Friedman E, Verderame M, Winawer S, Pollack R. Actin cytoskeletal organization loss in the benign-to-malignant tumor transition in cultured human colonic epithelial cells. Cancer Res 1984: 44: 3040. Tanaka H, Mori Y, Ishii H, Akedo H. Enhancement of metastatic capacity of fibroblast-tumor cell interaction in mice. Cancer Res 1988: 48: 1456. Talmadge JE, Fidler IJ. Enhanced metastatic potential of tumor cells harvested from spontaneous murine tumors. J Natl Cancer Inst 1982: 69: 975. Leung MF, Sokoloski JA, Sartorelli AC. Changes in microtubules, microtubule-associated proteins, and intermediate filaments during the differentiation of HL-60 leukemia cells. Cancer Res 1992: 52: 949. Bernal SD, Stahel RA. Cytoskeleton associated proteins; their role as cellular integrators in the neoplastic process. CRC Grit Rev Oncol Hematol 1985: 3: 191. Horio T, Hotani H. Visualization of the dynamic instability of individual microtubules by dark field microscopy. Nature 1986: 321: 605. Koch GLE, Macer DRJ, Smith MJ. Visualization of the intact endoplasmic reticulum by immunofluorescence with antibodies to the major ER glycoprotein, endoplasmin.. Cell Sci 1987: 87: 535. Bernal SD, Chen LB. Induction of cytoskeleton-associated proteins during differentiation of human myeloid leukemic cell lines. Cancer Res 1982: 42: 5106. Pollack R, Osborn M, Weber K. Patterns of organization of actin and myosin in normal and transformed cultured cells. Proc Nat Acad Sci 1975: 72: 994. Fink-Puches R, Hofmann-Wellenhof R, Smolle J, Kerl H. Confocal laser scanning microscopy - a new optical microscopic technique in pathology and dermatology. J Cutan Pathol 1995: 22: 252. White JG, Amos WB, Fordham M. An evaluation of confocal versus conventional imaging of biological structure by fluorescence light microscopy. J Cell Biol 1987: 105: 41. Helige C, Smolle J, Zellnig G, Fink-Puches R, Kerl H, Tritthart HA. Effect of dequalinium on K1735-M2 melanoma cell growth, directional migration and invasion in vitro. Eur J Cancer 1993: 29A, 1: 121. Smolle J, Stolz W, Balmier FA et al. Analytic morphology in clinical and experimental dermatology. J Am Acad Dermatol 1993: 29: 86. Humphries MJ, Yamada KM, Olden K. Investigation of the biological effects of anti-cell adhesive synthetic peptides that inhibit experimental metastasis of B16-F10 murine melanoma cells. J Clin Invest 1988: 81: 782. Brinkley BR, Fuller GM, Highfield DP. Cytoplasmic microtubules in normal and transformed cells in culture: Analysis by tubulin antibody immunofluorescence. Proc Nat Acad Sci 1975: 72: 4981. Grimslad IA. Direct evidence that cancer cell locomotion contributes importantly to invasion. Exp Cell Res 1987: 173: 515. Rungger-Brändle E, Gabbiani G. The role of cyloskeletal and cytocontractile elements in pathologic processes. Am J Pathol 1983: 110: 361. Carr I, Levy M, Watson P. The invasive edge: invasion in colorectal cancer. Clin Expl Metast 1986: 4: 129. Brown A, Li Y, Slaughter T, Black MM. Composite micro-tubules of the axon: quantitative analysis of tyrosinated and acetylated tubulin along individual axonal microtubules. J Cell Sci 1993: 104: 339. Hart IR, Raz A, Fidler IJ. Effect of cytoskeleton-disrupting agents on the metastatic behavior of melanoma cells. J Natl Cancer Inst 1980: 64: 891. Mareel M, De Brabander M. Effect of microtubule inhibitors on malignant invasion in vitro. J Nat Cancer Inst 1978: 61: 787. Kripke ML. Speculations on the role of the ultraviolet radiation in the development of malignant melanoma. J Natl Cancer Inst 1979: 63: 541. Mareel MM, Storme GA, DeBruyne GK, Van Cauwenberge RM. Vinblastine vincristine and vindesine: anti-invasive effect on MO4 mouse fibrosarcoma cells in vitro. Eur J Cancer Clin Oncol 1982: 18: 199. Starkey JR. Cell-matrix-interaction during tumor invasion. Cancer Metast Rev 1990: 9: 113. Smolle J, Soyer HP, Kerl H. Tubulin expression in melanocytic skin tumors - an immunohistochemical study. Am J Dermatopathol 1990: 12: 17. 1990; 12 1993; 29 1982; 18 1983; 110 1980; 64 1987; 105 1985; 3 1984; 44 1973 1975; 72 1993; 104 1992; 52 1982; 69 1987; 173 1987; 87 1993; 29A 1986; 321 1986; 4 1982; 42 1988; 48 1995; 22 1978; 61 1979; 63 1990; 9 1988; 81 Bernal SD (e_1_2_1_7_2) 1982; 42 Allison AC (e_1_2_1_27_2) 1973 e_1_2_1_22_2 e_1_2_1_23_2 e_1_2_1_21_2 Talmadge JE (e_1_2_1_10_2) 1982; 69 Friedman E (e_1_2_1_16_2) 1984; 44 Leung MF (e_1_2_1_24_2) 1992; 52 Brown A (e_1_2_1_25_2) 1993; 104 e_1_2_1_6_2 Koch GLE (e_1_2_1_20_2) 1987; 87 Mareel M (e_1_2_1_26_2) 1978; 61 Hart IR (e_1_2_1_17_2) 1980; 64 e_1_2_1_4_2 e_1_2_1_5_2 e_1_2_1_11_2 e_1_2_1_3_2 e_1_2_1_12_2 Tanaka H (e_1_2_1_13_2) 1988; 48 Helige C (e_1_2_1_14_2) 1993; 29 e_1_2_1_15_2 Rungger‐Brändle E (e_1_2_1_2_2) 1983; 110 e_1_2_1_19_2 e_1_2_1_8_2 e_1_2_1_9_2 e_1_2_1_18_2 |
| References_xml | – reference: Carr I, Levy M, Watson P. The invasive edge: invasion in colorectal cancer. Clin Expl Metast 1986: 4: 129. – reference: Leung MF, Sokoloski JA, Sartorelli AC. Changes in microtubules, microtubule-associated proteins, and intermediate filaments during the differentiation of HL-60 leukemia cells. Cancer Res 1992: 52: 949. – reference: Rungger-Brändle E, Gabbiani G. The role of cyloskeletal and cytocontractile elements in pathologic processes. Am J Pathol 1983: 110: 361. – reference: Kripke ML. Speculations on the role of the ultraviolet radiation in the development of malignant melanoma. J Natl Cancer Inst 1979: 63: 541. – reference: Hart IR, Raz A, Fidler IJ. Effect of cytoskeleton-disrupting agents on the metastatic behavior of melanoma cells. J Natl Cancer Inst 1980: 64: 891. – reference: White JG, Amos WB, Fordham M. An evaluation of confocal versus conventional imaging of biological structure by fluorescence light microscopy. J Cell Biol 1987: 105: 41. – reference: Helige C, Smolle J, Zellnig G, Fink-Puches R, Kerl H, Tritthart HA. Effect of dequalinium on K1735-M2 melanoma cell growth, directional migration and invasion in vitro. Eur J Cancer 1993: 29A, 1: 121. – reference: Smolle J, Soyer HP, Kerl H. Tubulin expression in melanocytic skin tumors - an immunohistochemical study. Am J Dermatopathol 1990: 12: 17. – reference: Bernal SD, Chen LB. Induction of cytoskeleton-associated proteins during differentiation of human myeloid leukemic cell lines. Cancer Res 1982: 42: 5106. – reference: Bernal SD, Stahel RA. Cytoskeleton associated proteins; their role as cellular integrators in the neoplastic process. CRC Grit Rev Oncol Hematol 1985: 3: 191. – reference: Mareel MM, Storme GA, DeBruyne GK, Van Cauwenberge RM. Vinblastine vincristine and vindesine: anti-invasive effect on MO4 mouse fibrosarcoma cells in vitro. Eur J Cancer Clin Oncol 1982: 18: 199. – reference: Mareel M, De Brabander M. Effect of microtubule inhibitors on malignant invasion in vitro. J Nat Cancer Inst 1978: 61: 787. – reference: Brown A, Li Y, Slaughter T, Black MM. Composite micro-tubules of the axon: quantitative analysis of tyrosinated and acetylated tubulin along individual axonal microtubules. J Cell Sci 1993: 104: 339. – reference: Koch GLE, Macer DRJ, Smith MJ. Visualization of the intact endoplasmic reticulum by immunofluorescence with antibodies to the major ER glycoprotein, endoplasmin.. Cell Sci 1987: 87: 535. – reference: Grimslad IA. Direct evidence that cancer cell locomotion contributes importantly to invasion. Exp Cell Res 1987: 173: 515. – reference: Friedman E, Verderame M, Winawer S, Pollack R. Actin cytoskeletal organization loss in the benign-to-malignant tumor transition in cultured human colonic epithelial cells. Cancer Res 1984: 44: 3040. – reference: Humphries MJ, Yamada KM, Olden K. Investigation of the biological effects of anti-cell adhesive synthetic peptides that inhibit experimental metastasis of B16-F10 murine melanoma cells. J Clin Invest 1988: 81: 782. – reference: Talmadge JE, Fidler IJ. Enhanced metastatic potential of tumor cells harvested from spontaneous murine tumors. J Natl Cancer Inst 1982: 69: 975. – reference: Pollack R, Osborn M, Weber K. Patterns of organization of actin and myosin in normal and transformed cultured cells. Proc Nat Acad Sci 1975: 72: 994. – reference: Brinkley BR, Fuller GM, Highfield DP. Cytoplasmic microtubules in normal and transformed cells in culture: Analysis by tubulin antibody immunofluorescence. Proc Nat Acad Sci 1975: 72: 4981. – reference: Horio T, Hotani H. Visualization of the dynamic instability of individual microtubules by dark field microscopy. Nature 1986: 321: 605. – reference: Starkey JR. Cell-matrix-interaction during tumor invasion. 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| SubjectTerms | Animals Biological and medical sciences Female Fluorescent Antibody Technique, Indirect Image Processing, Computer-Assisted Investigative techniques, diagnostic techniques (general aspects) Medical sciences Melanoma - metabolism Melanoma - ultrastructure Mice Mice, Inbred C3H Microscopy, Confocal Microtubules - drug effects Microtubules - metabolism Microtubules - ultrastructure Miscellaneous. Technology Neoplasm Metastasis - ultrastructure Nocodazole - pharmacology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Tumor Cells, Cultured |
| Title | Cytoplasmic microtubules in two different mouse melanoma cell lines: a qualitative and quantitative analysis using confocal laser scanning microscopy and computer-assisted image analysis |
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