Abstract
The effect of X‐irradiation on production of MUC1 was studied with human colon carcinoma HT‐29 cells. As evaluated by immunocytochemical staining, the percentages of MUC1‐positive cells in cells at 4 days after 6 Gy irradiation and in unirradiated control cells were 52±3.5% (n=6) and 26±2.8% (n=6), respectively. Flow‐cytometric analysis of living cells showed that MUC1 began to rise from day 1, reaching a plateau by day 4 after 6 Gy irradiation. Western blot analysis with monoclonal antibody MY.1E12 against glycosylated MUC1 (mature form) showed dose‐dependent increases of two bands (500 and 390 kDa) corresponding to two polymorphic MUC1 alleles. Premature forms of MUC1 (350 and 240 kDa) were detectable with monoclonal antibody HMFG‐2 only in irradiated cells, suggesting that new core protein synthesis had been induced. The transcriptional activity of the MUC1 gene was analyzed in terms of transient expression of MUC1‐CAT reporter plasmids containing 5′‐flanking sequences of the MUC1 gene fused to the bacterial chloramphenicol acetyltransferase (CAT) gene. The results of CAT assay indicate that enhanced expression of MUC1 in irradiated HT‐29 cells was due to upregulation of MUC1 transcription, and required the upstream promoter.
Keywords: MUC1, Radiation, Colon carcinoma cell, Expression, Tumor marker
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REFERENCES
- 1. ) Hilkens , J. , Buijs , F. , Hilgers , J. , Hageman , P. , Calafat , J. , Sonnenberg , A. and van der Valk , M.Monoclonal antibodies against human milk‐fat globule membranes detecting differentiation antigens of the mammary gland and its tumors . Int. J. Cancer , 34 , 197 – 206 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Nakamori , S. , Ota , D. M. , Cleary , K. R. , Shirotani , K. and Irimura , T.MUC1 mucin expression as a marker of progression and metastasis of human colorectal carcinoma . Gastroenterology , 106 , 353 – 361 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Irimura , T. , Carlson , D. A. , Price , J. , Yamori , T. , Giavazzi , R. , Ota , D. M. and Cleary , K. R.Differential expression of a sialoglycoprotein with an approximate molecular weight of 900,000 on metastatic human colon carcinoma cells growing in culture and in tumor tissues . Cancer Res. , 48 , 2353 – 2360 ( 1988. ). [PubMed] [Google Scholar]
- 4. ) Girling , A. , Bartkova , J. , Burchell , J. , Gendler , S. , Gillett , C. and Taylor‐Papadimitriou , J.A core protein epitope of the polymorphic epithelial mucin detected by the monoclonal antibody SM‐3 is selectively exposed in a range of primary carcinomas . Int. J. Cancer , 43 , 1072 – 1076 ( 1989. ). [DOI] [PubMed] [Google Scholar]
- 5. ) Burchell , J. , Gendler , S. , Taylor‐Papadimitriou , J. , Girling , A. , Lewis , A. , Millis , R. and Lamport , D.Development and characterization of breast cancer reactive monoclonal antibodies directed to the core protein of the human milk mucin . Cancer Res. , 47 , 5476 – 5482 ( 1987. ). [PubMed] [Google Scholar]
- 6. ) Lloyd , K. O. , Burchell , J. , Kudryashov , V. , Yin , B. W. T. and Taylor‐Papadimitriou , J.Comparison of O‐linked carbohydrate chains in MUC‐1 mucin from normal breast epithelial cell lines and breast carcinoma cell lines. Demonstration of simpler and fewer glycan chains in tumor cells . J. Biol. Chem. , 271 , 33325 – 33334 ( 1996. ). [DOI] [PubMed] [Google Scholar]
- 7. ) Makiguchi , Y. , Hinoda , Y. and Imai , K.Effect of MUC1 mucin, an anti‐adhesion molecule, on tumor cell growth . Jpn. J. Cancer Res. , 87 , 505 – 511 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. ) Wesseling , J. , van der Valk , S. W. and Hilkens , J.A mechanism for inhibition of E‐cadherin‐mediated cell‐cell adhesion by the membrane‐associated mucin episialin/MUC1 . Mol. Biol. Cell , 7 , 565 – 577 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. ) Wesseling , J. , van der Valk , S. W. , Vos , H. L. , Sonnenberg , A. and Hilkens , J.Episialin (MUC1) overexpression inhibits integrin‐mediated cell adhesion to extracellular matrix components . J. Cell Biol. , 129 , 255 – 265 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. ) Suzuki , N.Centrifugal elutriation and characterization of tumor cells from venous blood of tumor‐bearing mice: possible relevance to metastasis . Cancer Res. , 44 , 3505 – 3511 ( 1984. ). [PubMed] [Google Scholar]
- 11. ) Suzuki , N.New method to quantitate clonogenic tumor cells in the blood circulation of mice . Cancer Res. , 43 , 5451 – 5455 ( 1983. ). [PubMed] [Google Scholar]
- 12. ) Morimatsu , A. , Suzuki , N. , Hirano , K. , Matsumoto , Y. and Sakai , K.Identification and characterization of a protein found after X‐irradiation in human T cell leukemia . J. Radiat. Res. (Tokyo) , 37 , 1 – 11 ( 1996. ). [DOI] [PubMed] [Google Scholar]
- 13. ) Irimura , T. , McIssac , A. M. , Carlson , D. A. , Yagita , M. , Grimm , E. A. , Menter , D. G. , Ota , D. M. and Cleary , K. R.Soluble factor in normal tissues that stimulates high‐molecular‐weight sialoglycoprotein production by human colon carcinoma cells . Cancer Res. , 50 , 3331 – 3338 ( 1990. ). [PubMed] [Google Scholar]
- 14. ) Dohi , D. F. , Sutton , R. C. , Frazier , M. L. , Nakamori , S. , McIsaac , A. M. and Irimura , T.Regulation of sialomucin production in colon carcinoma cells . J. Biol. Chem. , 268 , 10133 – 10138 ( 1993. ). [PubMed] [Google Scholar]
- 15. ) Shirotani , K. , Taylor‐Papadimitriou , J. , Gendler , S. J. and Irimura , T.Transcriptional regulation of the MUC1 mucin gene in colon carcinoma cells by a soluble factor . J. Biol. Chem. , 269 , 15030 – 15035 ( 1994. ). [PubMed] [Google Scholar]
- 16. ) Shirotani , K. and Irimura , T.Purification of nuclear proteins that potentially regulate transcription of the MUC1 mucin gene induced by a soluble factor . J. Biochem. , 124 , 585 – 590 ( 1998. ). [DOI] [PubMed] [Google Scholar]
- 17. ) Lesuffleur , T. , Porchet , N. , Aubert , J. P. , Swallow , D. , Gum , J. R. , Kim , Y. S. , Real , F. X. and Zweibaum , A.Differential expression of the human mucin genes MUC1 to MUC5 in relation to growth and differentiation of different mucus‐secreting HT‐29 cell subpopulations . J. Cell Sci. , 106 , 771 – 783 ( 1993. ). [DOI] [PubMed] [Google Scholar]
- 18. ) Yamamoto , M. , Bhavanandan , V. P. , Nakamori , S. and Irimura , T.A novel monoclonal antibody specific for sialylated MUC1 mucin . Jpn. J. Cancer Res. , 87 , 488 – 496 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19. ) Ho , J. J. , Cheng , S. and Kim , Y. S.Access to peptide regions of a surface mucin (MUC1) is reduced by sialic acids . Biochem. Biophys. Res. Commun. , 210 , 866 – 873 ( 1995. ). [DOI] [PubMed] [Google Scholar]
- 20. ) Lancaster , C. A. , Peat , N. , Duhig , T. , Wilson , D. , Taylor‐Papadimitriou , J. and Gendler , S. J.Structure and expression of the human polymorphic epithelial mucin gene: an expressed VNTR unit . Biochem. Biophys. Res. Commun. , 173 , 1019 – 1029 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 21. ) Kovarik , A. , Peat , N. , Wilson , D. , Gendler , S. J. and Taylor‐Papadimitriou , J.Analysis of the tissue‐specific promoter of the MUC1 gene . J. Biol. Chem. , 268 , 9917 – 9926 ( 1993. ). [PubMed] [Google Scholar]
- 22. ) Hilkens , J. and Buijs , F.Biosynthesis of MAM‐6, an epithelial sialomucin . J. Biol. Chem. , 263 , 4215 – 4222 ( 1988. ). [PubMed] [Google Scholar]
- 23. ) Litvinov , S. V. and Hilkens , J.The epithelial sialomucin, episialin, is sialylated during recycling . J. Biol. Chem. , 268 , 21364 – 21371 ( 1993. ). [PubMed] [Google Scholar]
- 24. ) Sawaya , R. , Tofilon , P. J. , Mohanam , S. , Ali‐Osman , F. , Liotta , L. A. , Stetler‐Stevenson , W. G. and Rao , J. S.Induction of tissue‐type plasminogen activator and 72‐kDa type‐IV collagenase by ionizing radiation in rat astrocytes . Int. J. Cancer , 56 , 214 – 218 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 25. ) Boothman , D. A. , Wang , M. and Lee , S. W.Induction of tissue‐type plasminogen activator by ionizing radiation in human malignant melanoma cells . Cancer Res. , 51 , 5587 – 5595 ( 1991. ). [PubMed] [Google Scholar]
- 26. ) Stein , B. , Rahmsdorf , H. J. , Steffen , A. , Litfin , M. and Herrlich , P.UV‐induced DNA damage is an intermediate step in UV‐induced expression of human immunodeficiency virus type 1, collagenase, c‐fos, and metallothionein . Mol. Cell. Biol. , 9 , 5169 – 5181 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27. ) Lan , M. S. , Batra , S. K. , Qi , W. N. , Metzgar , R. S. and Hollingsworth , M. A.Cloning and sequencing of a human pancreatic tumor mucin cDNA . J. Biol. Chem. , 265 , 15294 – 15299 ( 1990. ). [PubMed] [Google Scholar]
- 28. ) Ligtenberg , M. J. , Vos , H. L. , Gennissen , A. M. and Hilkens , J.Episialin, a carcinoma‐associated mucin, is generated by a polymorphic gene encoding splice variants with alternative amino termini . J. Biol. Chem. , 265 , 5573 – 5578 ( 1990. ). [PubMed] [Google Scholar]
- 29. ) Sahijdak , W. M. , Yang , C. R. , Zuckerman , J. S. , Meyers , M. and Boothman , D. A.Alterations in transcription factor binding in radioresistant human melanoma cells after ionizing radiation . Radiat. Res. , 138 , S47 – S51 ( 1994. ). [PubMed] [Google Scholar]