Cancer Letters
Volume 194, Issue 2 , Pages 199-208 , 15 May 2003

Human epithelial cell immortalization as a step in carcinogenesis

Received 10 September 2002 ,Revised 29 October 2002 ,Accepted 29 October 2002.

References 

  1. Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PLC, et al. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994;266:2011–2015
  2. Ducrest AL, Szutorisz H, Lingner J, Nabholz M. Regulation of the human telomerase reverse transcriptase gene. Oncogene. 2002;21:541–552
  3. Collins K, Mitchell JR. Telomerase in the human organism. Oncogene. 2002;21:564–579
  4. Stampfer MR, Bartley JC. Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo(a)pyrene. Proc. Natl. Acad. Sci. USA. 1985;82:2394–2398
  5. Wazer DE, Chu Q, Liu X-L, Gao Q, Safaii H, Band V. Loss of p53 during radiation transformation of primary human mammary epithelial cells. Mol. Cell Biol. 1994;14:2468–2478
  6. Nonet G, Stampfer MR, Chin K, Gray JW, Collins CC, Yaswen P. The ZNF217 gene amplified in breast cancers promotes immortalization of human mammary epithelial cells. Cancer Res. 2001;61:1250–1254
  7. Li Z, Meng ZH, Sayeed A, Shalaby R, Ljung B-M, Dairkee SH. Genome wide allelotyping of a new in vitro model system reveals early events in breast cancer progression. Cancer Res. 2002;62:5980–5987
  8. Band V, Zajchowski D, Kulesa V, Sager R. Human papilloma virus DNAs immortalize normal human mammary epithelial cells and reduce their growth factor requirements. Proc. Natl. Acad. Sci. USA. 1990;87:463–467
  9. Shay JW, Wright WE, Brasiskyte D, Van Der Haegen BA. E6 of human papillomavirus type 16 can overcome the M1 stage of immortalization in human mammary epithelial cells but not in human fibroblasts. Oncogene. 1993;8:1407–1413
  10. Wazer DE, Liu X-L, Chu Q, Gao Q, Band V. Immortalization of distinct human mammary epithelial cell types by human papilloma virus 16 E6 or E7. Proc. Natl. Acad. Sci. USA. 1995;92:3687–3691
  11. Sherr CJ, De Pinho RA. Cellular senescence: mitotic clock or culture shock?. Cell. 2000;102:407–410
  12. Serrano M, Blasco MA. Putting the stress on senescence. Cur. Opin. Cell Biol. 2001;13:748–753
  13. Dimri GP, Lee X, Basile G, Roskelley C, Medrano EE, Rubelji I, et al. A novel biomarker identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl. Acad. Sci. USA. 1995;92:9363–9367
  14. Romanov S, Kozakiewicz K, Holst C, Stampfer MR, Haupt LM, Tlsty T. Normal human mammary epithelial cells spontaneously escape senescence and acquire genomic changes. Nature. 2001;409:633–637
  15. Prowse KR, Greider CW. Developmental and tissue-specific regulation of mouse telomerase and telomere length. Proc. Natl. Acad. Sci. USA. 1995;92:4818–4822
  16. Greenberg RA, Allsopp RC, Chin L, Morin GB, De Pinho RA. Expression of mouse telomerase reverse transcriptase during development, differentiation and proliferation. Oncogene. 1998;16:1723–1730
  17. Pierce JH, Arnstein P, DiMarco E, Artrip J, Kraus MH, Lonardo F, et al. Oncogenic potential of erbB-2 in human mammary epithelial cells. Oncogene. 1991;6:1189–1194
  18. Frittitta L, Vigneri R, Stampfer MR, Goldfine ID. Insulin receptor overexpression in 184B5 human mammary epithelial cells induces a ligand-dependent transformed phenotype. J. Cell. Biochem. 1995;57:666–669
  19. Olsen CL, Gardie B, Yaswen P, Stampfer MR. Raf-1-induced growth arrest in human mammary epithelial cells is p16-independent and is overcome in immortal cells during conversion. Oncogene. 2002;21:6328–6339
  20. Wright WE, Pereira-Smith OM, Shay JW. Reversible cellular senescence: implications for immortalization of normal human diploid fibroblasts. Mol. Cell. Biol. 1989;9:3088–3092
  21. Shay JW, Wright WE, Werbin H. Toward a molecular understanding of human breast cancer: a hypothesis. Breast Cancer Res. Treat. 1993;25:83–94
  22. Wright WE, Shay JW. Time, telomeres and tumors: is cellular senescence more than an anticancer mechanism. Trends Cell Biol. 1995;5:293–297
  23. von Zglinicki T. Oxidative stress shortens telomeres. Trends Biochem. Sci. 2002;27:339–344
  24. Loughran O, Malliri A, Owens D, Gallimore PH, Stanley MA, Ozanne B, et al. Association of CDKN2A/p16INK4A with human head and neck keratinocyte replicative senescence: relationship of dysfunction to immortality and neoplasia. Oncogene. 1996;13:561–568
  25. Brenner AJ, Stampfer MR, Aldaz CM. Increased p16INK4a expression with onset of senescence of human mammary epithelial cells and extended growth capacity with inactivation. Oncogene. 1998;17:199–205
  26. Jarrard DF, Sarkar S, Shi Y, Yeager TR, Magrane G, Kinoshita H, et al. p16/pRb pathway alterations are required for bypassing senescence in human prostate epithelial cells. Cancer Res. 1999;59:2957–2964
  27. Sandhu C, Peehl DM, Slingerland J. p16INK4A mediates cyclin dependent kinase 4 and 6 inhibition in senescent prostatic epithelial cells. Cancer Res. 2000;60:2616–2622
  28. Dickson MA, Hahn WC, Ino Y, Ronfard V, Wu JY, Weinberg RA, et al. Human keratinocytes that express hTERT and also bypass a p16INK4a-enforced mechanism that limits lifespan become immortal yet retain normal growth and differentiation characteristics. Mol. Cell Biol. 2000;20:1436–1447
  29. Rheinwald JG, Hahn WC, Ramsey MR, Wu JY, Guo Z, Tsao H, et al. A two-stage p16INK4a-and p53-dependent keratinocyte senescence mechanism that limits replicative potential independent of telomere status. Mol. Cell. Biol. 2002;22:5157–5172
  30. Bringold F, Serrano M. Tumor suppressors and oncogenes in cellular senescence. Exp. Gerontol. 2000;35:317–329
  31. Sandhu C, Donovan J, Bhattacharya N, Stampfer M, Worland P, Slingerland J. Reduction of Cdc25A contributes to cyclin E1-Cdk2 inhibition at senescence in human mammary epithelial cells. Oncogene. 2000;19:5314–5323
  32. Brown JP, Wei W, Sedivy JM. Bypass of senescence after disruption of p21CIP1/WAF1 gene in normal diploid human fibroblasts. Science. 1997;277:831–834
  33. Stampfer MR, Yaswen P. Immortal transformation and telomerase reactivation of human mammary epithelial cells in culture. In:  Mattson M,  Pandita T editor. Telomerase, Aging and Disease. Vol. 8:Amsterdam: Elsevier; 2001;p. 103–130
  34. Ramirez RD, Morales CP, Herbert BS, Rohde JM, Passons C, Shay JW, et al. Putative telomere-independent mechanisms of replicative aging reflect inadequate growth conditions. Genes Dev. 2001;15:398–403
  35. Stampfer M, Garbe J, Levine G, Lichsteiner S, Vasserot A, Yaswen P. Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor β growth inhibition in p16INK4 (−) human mammary epithelial cells. Proc. Natl. Acad. Sci. USA. 2001;98:4498–4503
  36. T'Ang A, Varley JM, Chakraborty S, Murphree AL, Fung Y-KT. Structural rearrangement of the retinoblastoma gene in human breast carcinoma. Science. 1988;242:263–266
  37. Landberg G, Nielson NH, Nilsson P, Emdin SO, Cajander J, Roos G. Telomerase activity is associated with cell cycle deregulation in human breast cancer. Cancer Res. 1997;57:549–554
  38. Band V. The role of retinoblastoma and p53 tumor suppressor pathways in human mammary epithelial cell immortalization. Int. J. Oncol. 1998;12:499–507
  39. Geradts J, Wilson PA. High frequency of aberrant p16INK4A expression in human breast cancer. Am. J. Pathol. 1996;149:15–20
  40. Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP. Alterations in DNA methylation: a fundamental aspect of neoplasia. Adv. Cancer Res. 1998;72:141–196
  41. Huschtscha LI, Noble JR, Neumann AA, Moy EL, Barry P, Melki JR, et al. Loss of p16INK4 expression by methylation is associated with lifespan extension of human mammary epithelial cells. Cancer Res. 1998;58:3508–3512
  42. Foster SA, Wong DJ, Barrett MT, Galloway DA. Inactivation of p16 in human mammary epithelial cells by CpG island methylation. Mol. Cell. Biol. 1998;18:1793–1801
  43. Bartkova J, Lukas J, Muller H, Lutzhoft D, Strauss M, Bartek J. Cyclin D1 protein expression and function in human breast cancer. Int. J. Cancer. 1994;57:353–361
  44. Weinstat-Saslow D, Merino M, Manrow R, Lawrence J, Bluth R, Wittenbel K, et al. Overexpression of cyclin D mRNA distinguishes invasive and in situ breast carcinomas from non-malignant lesions. Nat. Med. 1995;1:1257–1260
  45. Loden M, Stighall M, Nielson NH, Roos G, Emdin SO, Ostlund H, et al. The cyclin D1 high and cyclin E high subgroups of breast cancer: separate pathways in tumorigenesis based on patterns of genetic alterations and inactivation of the pRb node. Oncogene. 2002;21:4680–4690
  46. Wolfel T, Hauer M, Schneider J, Serrano M, Wolfel C, Klehmann-Hieb E, et al. A p16INK4a-insensitive CDK4 mutant targeted by cytolytic T lymphocytes in a human melanoma. Science. 1995;269:1281–1284
  47. An HX, Beckmann MW, Reifenberger G, Bender HG, Niederacher D. Gene amplification and overexpression of CDK4 in sporadic breast carcinomas is associated with high tumor cell proliferation. Am. J. Pathol. 1999;154:113–118
  48. Jacobs JJ, Kieboom K, Marino S, De Pinho RA, van Lohuizen M. The oncogene and Polycomb-group gene bmi-1 regulates cell proliferation and senescence through the ink4a locus. Nature. 1999;397:164–168
  49. Zebedee Z, Hara E. Id proteins in cell cycle control and cellular senescence. Oncogene. 2001;20:8317–8325
  50. Drayton S, Peters G. Immortalisation and transformation revisited. Curr. Opin. Genet. Dev. 2002;12:98–104
  51. Wright WE, Shay JW. Historical claims and current interpretations of replicative aging. Nat. Biotechnol. 2002;20:682–688
  52. Shay JW, Van Der Haegen BA, Ying Y, Wright WE. The frequency of immortalization of human fibroblasts and mammary epithelial cells transfected with SV40 Large T-antigen. Exp. Cell Res. 1993;209:45–52
  53. Tlsty TD, Romanov SR, Kozakiewicz BK, Holst CR, Haupt LM, Crawford YG. Loss of chromosomal integrity in human mammary epithelial cells subsequent to escape from senescence. J. Mammary Gland Biol. Neoplasia. 2001;6:235–243
  54. Foster SA, Galloway DA. Human papillomavirus type 16 E7 alleviates a proliferative block in early passage human mammary epithelial cells. Oncogene. 1996;12:1773–1779
  55. Shay JW, Tomlinson G, Piatyszek MA, Gollahon LS. Spontaneous in vitro immortalization of breast epithelial cells from a patient with Li-Fraumeni syndrome. Mol. Cell. Biol. 1995;15:425–432
  56. Gollahon LS, Shay JW. Immortalization of human mammary epithelial cells transfected with mutant p53 (273his). Oncogene. 1996;12:715–725
  57. Gao Q, Hauser SH, Liu X-L, Wazer DE, Madoc-Jones H, Band V. Mutant p53-induced immortalization of primary human mammary epithelial cells. Cancer Res. 1996;56:3129–3133
  58. Van Der Haegen BA, Shay JW. Immortalization of human mammary epithelial cells by SV40 large T-antigen involves a two step mechanism. In Vitro Cell. Dev. Biol. 1993;29A:180–182
  59. Bartek J, Bartkova J, Kyprianou N, Lalani E-N, Staskova Z, Shearer M, et al. Efficient immortalization of luminal epithelial cells from human mammary gland by introduction of simian virus 40 large tumor antigen with a recombinant retrovirus. Proc. Natl. Acad. Sci. USA. 1991;88:3520–3524
  60. Counter CM, Avilion AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB, et al. Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J. 1992;11:1921–1929
  61. Wei S, Wei W, Sedivy JM. Expression of catalytically active telomerase does not prevent premature senescence caused by overexpression of oncogenic Ha-Ras in normal human fibroblasts. Cancer Res. 1999;59:1539–1543
  62. Klingelhutz AJ, Foster SA, McDougall JK. Telomerase activation by the E6 gene product of human papillomavirus type 16. Nature. 1996;380:79–82
  63. Kiyono T, Foster SA, Koop JJ, McDougall JK, Galloway DA, Klingelhutz AJ. Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cell. Nature. 1998;396:84–88
  64. Härle-Bachor C, Boukamp P. Telomerase activity in the regenerative basal layer of the epidermis in human skin and in immortal and carcinoma-derived skin keratinocytes. Proc. Natl. Acad. Sci. USA. 1996;93:6476–6481
  65. Alani RM, Hasskarl J, Grace M, Hernandez M-C, Israel M, Munger K. Immortalization of primary human keratinocytes by the helix-loop-helix protein, Id-1. Proc. Natl. Acad. Sci. USA. 1999;96:9637–9641
  66. Dellambra E, Golisano O, Bondanza S, Siviero E, Lacal P, Molinari M, et al. Downregulation of 14-3-3σ prevents clonal evolution and leads to immortalization of primary human keratinocytes. J. Cell Biol. 2000;149:1117–1129
  67. Tsao J, Yanle Z, Lukas J, Yang X, Shah A, Press M, et al. Telomerase activity in normal and neoplastic breast. Clin. Cancer Res. 1997;3:627–631
  68. Mokbell K, Parris CN, Ghilchik M, Newbold RF. Telomerase activity in the human breast. Breast. 1999;8:208–211
  69. Garbe J, Wong M, Wigington D, Yaswen P, Stampfer MR. Viral oncogenes accelerate conversion to immortality of cultured human mammary epithelial cells. Oncogene. 1999;18:2169–2180
  70. Wang J, Hannon GJ, Beach DH. Risky immortalization by telomerase. Nature. 2000;405:755–756
  71. M.R. Stampfer, J. Garbe, D. Wigington, T. Nijjar, K. Swisshelm, P. Yaswen, Rapid conversion to telomerase(+) full immortality of indefinite lifespan human mammary epithelial cell lines lacking functional p53. submitted.
  72. Lehman T, Modali R, Boukamp P, Stanek J, Bennett W, Welsh J, et al. p53 mutations in human immortalized epithelial cell lines. Carcinogenesis. 1993;14:833–839
  73. Stampfer MR, Bodnar A, Garbe J, Wong M, Pan A, Villeponteau B, et al. Gradual phenotypic conversion associated with immortalization of cultured human mammary epithelial cells. Mol. Biol. Cell. 1997;8:2391–2405
  74. Nijjar T, Wigington D, Garbe JC, Waha A, Stampfer MR, Yaswen P. p57/KIP2 loss of heterozygosity and expression during immortal conversion of human mammary epithelial cells. Cancer Res. 1999;59:5112–5118
  75. Walen K, Stampfer MR. Chromosome analyses of human mammary epithelial cells at stages of chemically-induced transformation progression to immortality. Cancer Genet. Cytogenet. 1989;37:249–261
  76. Li H, Cao Y, Berndt MC, Funder JW, Liu J-P. Molecular interactions between telomerase and the tumor suppressor protein p53 in vitro. Oncogene. 1999;18:6785–6794
  77. Xu D, Wang Q, Gruber A, Bjorkholm M, Chen Z, Zaid A, et al. Downregulation of telomerase reverse transcriptase mRNA expression by wild-type p53 in human tumor cells. Oncogene. 2000;19:5123–5133
  78. Kanaya T, Kyo S, Hamada K, Takakura M, Kitagawa Y, Harada H, et al. Adenoviral expression of p53 represses telomerase activity through down-regulation of human telomerase reverse transcriptase transcription. Clin Cancer Res. 2000;6:1239–1247
  79. Nielsen GP, Stemmer-Rachamimov AO, Shaw J, Roy JE, Koh J, Louis DN. Immunohistochemical survey of p16INK4A expression in normal human adult and infant tissues. Lab. Invest. 1999;79:1137–1143
  80. Brash DE, Rudolph JA, Simon JA, Lin A, McKenna GJ, Baden HP, et al. for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. Proc. Natl. Acad. Sci. USA. 1991;88:10124–10128
  81. Sjogren S, Inganas M, Norberg T, Lindgren A, Nordgren H, Holmberg L, et al. The p53 gene in breast cancer: prognostic value of complementary DNA sequencing versus immunohistochemistry.. J. Natl. Cancer Inst. 1996;88:173–182
  82. Roos G, Nilsson P, Cajander S, Nielsen N-H, Arnerlöv C, Landberg G. Telomerase activity in relation to p53 status and clinico-pathological parameters in breast cancer. Int. J. Cancer. 1998;79:343–348
  83. Iacopetta B, Grieu F, Powell B, Soong R, McCaul K, Seshadri R. Analysis of p53 gene mutation by polymerase chain reaction-single strand conformation polymorphism provides independent prognostic information in node-negative breast cancer. Clin. Cancer Res. 1998;4:1597–1602

PII: S0304-3835(02)00707-3

doi: 10.1016/S0304-3835(02)00707-3

Cancer Letters
Volume 194, Issue 2 , Pages 199-208 , 15 May 2003