臺灣博碩士論文加值系統

細胞週期進行是受到cyclins、 cyclin-depedent kinase 與cyclin-depedent kinase inhibitor 所調控, 而端粒酵素被認為可能與細胞週期進行有關。 hTERT催化次單位是端粒酵素的組成之一, 其表現被認為與腫瘤形成有關, 雖然關於調控hTERT表現的機制並不清楚, 但仍有許多研究認為c-myc可以活化hTERT基因並使其表現, 除此之外, c-myc可受到細胞週期上游的調控因子p16與cyclin D1/CDK4所調控, 而影響c-myc在基因轉錄及細胞轉形的作用。 因此本實驗希望了解膀胱癌中細胞週期調控因子對端粒酵素活性調控之關係及與臨床病理分期分級是否具有相關性。
本實驗共收集65個膀胱癌組織, 藉TRAP-ELISA方法分析端粒酵素活性, 利用RT-PCR方法分析hTERT mRNA表現, 以及利用西方墨點法分析cyclin D1、c-Myc以及P16等蛋白質表現。
結果在65個膀胱癌組織測到56個 (86%) 檢體其端粒酵素呈陽性, 59個檢體 (90%)有hTERT表現, 利用McNemar’s exact test 統計方法分析 (p>0.05), 發現端粒酵素活性及hTERT表現具有一致性的趨勢。此外, c-Myc蛋白質過度表現有57% (37/65)、 有37個 檢體(57%) 是P16蛋白質不表現或過度表現, 28個檢體 (43%) cyclin D1蛋白質過度表現。 在65個檢體中, 至少一種細胞週期調控因子不正常表現有60個佔92%。 結果顯示這些蛋白質正常表現與否對端粒酵素活性及hTERT mRNA的表現, 並無顯著相關性。 另外, 除cyclin D1外, P16、 c-Myc及hTERT正常表現與否與細胞分期分級無關, 然而cyclin D1蛋白質過度表現在侵犯性癌有較高發生的傾向 (p=0.058)。 本實驗發現利用分析端粒酵素組成hTERT 的表現對膀胱癌診斷之敏感度比偵測端粒酵素活性高, 除cyclin D1不正常表現與侵犯性癌較有關外, 細胞週期G1期調控因子與hTERT 的不正常表現認為可能在細胞癌化早期發生。 雖我們不能清楚的指出這些調控因子不正常表現與調節hTERT 表現的關係, 但仍認為累積這些不正常表現之調控因子可能是導致端粒酵素活性失控可能的原因, 以及導致膀胱癌癌化可能的重要因素。

Purpose: Telomerase is likely to have a correlation with cell cycle progression, which is controlled by the regulation of cyclins, CDK and CDKI. Expression of the telomearse catalytic sub-unit (hTERT) constitutes a key step in the development of human cancer. Although hTERT regulation is still unclear, several studies suggest that c-myc may activate its expression. Besides, cell cycle regulator p16 and cyclin D1/CDK4 complexes are upstream regulators of c-myc and directly govern c-myc function in transcriptional transactivation and transformation. The aims of this study were to investigate the involvements of these cell cycle regulators in telomerase activation in bladder carcinogenesis and correlate these findings with clinicopathological features.
Materials and Methods: Sixty-five bladder carcinomas were collected. TRAP-ELISA method was used to measure the telomerase activity. Expression of hTERT mRNA was measured by quantitative RT-PCR. Western blotting method was used to assess the protein expression of P16、cyclin D1 and c-Myc.
Results: Telomerase activity was detected in 56 (86%) of 65 bladder tumors, whereas telomerase evaluated by hTERT expression was observed in 59 tumors (90%). Significant concordance between telomerase activity and hTERT expression (McNemar’s exact test, p>0.05) was shown. c-Myc overexpression was seen in 57% (37/65) of tumors. Thirty-seven (57%) tumors lacked or overexpressed of P16 and 28 tumors (43%) overexpressed cyclin D1. Abnormal expression in at least one of these regulatory protein was seen in 60 (92%) of 65 tumors. There was no correlation between P16、 cyclin D1、 c-Myc and hTERT expression. No significant correlation between hTERT, P16、 c-Myc and the clinicopathologic factors was found. However, overexpression of cyclin D1 showed an association with invasive tumors (p = 0.058).
Discussion: The measurement of hTERT expression is more sensitive than telomerase activity assay to detect early bladder tumor. Except cyclin D1, abnormal expression of the G1 cell cycle regulators and hTERT expression may constitute an early event in tumor development. Though, we could not demonstrate a clear relation between each protein alterations and hTERT expression, but suggesting that aberrant accumulation of these proteins was considered as a reason for telomerase deregulation, which may play an essential role in bladder tumor pathogenesis.

英文摘要 1-3
中文摘要 4-5
前言 6-17
膀胱癌的發生機率與可能發生的原因 6-7
膀胱癌的症狀與分類 7-8
膀胱癌的診斷與分子生物學研究 8-10
端粒、端粒酵素與細胞生長週期G1期調控因子 10-17
材料與方法 18-35
(一). 細胞株與組織檢體之收集 18-19
(二). 端粒酵素活性之測定 20-25
(三). Total RNA萃取與RT-PCR(Reverse transcription- PCR)法偵測端粒酵素催化次單位hTERT mRNA之表現情形 25-29
(四). 西方墨點法(Western blotting)測定P16, cyclin D1, c-Myc蛋白質表現 29-35
結果 36-46
端粒酵素活性( telomerase activity ) 36-37
hTERT mRNA 表現 37-38
端粒酵素活性與hTERT mRNA 表現之比較 38-39
cyclin D1蛋白質的表現情形 39-40
c- Myc蛋白質的表現情形 40-41
P16蛋白質的表現情形 41-42
端粒酵素活化、hTERT mRNA表現與cyclin D1、c- Myc 、P16蛋白質的表現關係 42-44
cyclin D1、c-Myc 、P16蛋白質間相互的表現在腫瘤的分級、分期與端粒酵素活化情形的關係 44-46
討論 47-61
參考文獻 62-94
圖次 95-97
圖一: RNA電泳(18S & 28S) 95
圖二: 用RT-PCR分析hTERT mRNA的表現結果 96
圖三: 利用西方墨點法分析cyclin D1、c-Myc與P16蛋白質之表現情形 97
表次 98-108
表一: 膀胱癌腫瘤組織端粒酵素活性與hTERT mRNA表現 98
表二: Telomerase activity 、hTERT mRNA 、cyclin D1、c-MYC與P16蛋白質在膀胱癌的表現 99-100
表三: Telomerase activity 、hTERT mRNA 、cyclin D1、c-MYC與P16蛋白質在膀胱癌的表現 101-102
表四: 端粒酵素活性、hTERT mRNA、cyclin D1、c- Myc、P16在膀胱癌之細胞分期與分級的表現情形 103
表五: 端粒酵素活性在膀胱癌之細胞分期與分級的表現情形 104
表六: cyclin D1、c-Myc及P16蛋白質在膀胱癌腫瘤組織檢體之表現情形 105
表七: 端粒酵素活性與cyclin D1、c- Myc、P16蛋白質表現情形 106
表八: hTERT mRNA 表現、cyclin D1、c - Myc、P16蛋白質之間的表現情形 107
表九: 端粒酵素活性、腫瘤分期及分級與cyclin D1、c - Myc、P16蛋白質表現相互間正常與否在膀胱癌的表現情形 108
附圖次 109-118
附圖一: 染色體末端複製問題 109
附圖二: 端粒長度、端粒酵素活化與細胞生命期之相關性 110
附圖三: 端粒酵素活化之模式 111
附圖四: hTERT promoter 之構造 112
附圖五: c-Myc與影響細胞生長之下游調控因子的相關性 113
附圖六: c-Myc與細胞週期調控因子之相關性 114
附圖七:TRAP — ELISA Assay之原理 115
附圖八: semi- dry transfer 裝置 116
附圖九: ECL Western blotting 原理 117
附圖十:Homologous recombination 方式延長染色體末端端粒長度 118
附表次 119-120
附表一: 行政院衛生署九十年度衛生統計年報之罹患膀胱癌人數附表統計資料 119
附表二: 膀胱癌之細胞形態分類標準 120
附表三: 膀胱癌之細胞形態分類標準 121

Agarwala S, Hemal AK, Seth A, Gupta AK, Bhatnagar V, Mitra DK. (2001) Transitional cell carcinoma of the urinary bladder following exposure to cyclophosphamide in childhood. Eur J Pediatr Surg 11(3): 207-10.
Alcorta DA, Xiong Y, Phelps D, Hannon G, Beach D, Barrett JC. (1996) Involvement of the cyclin-dependent kinase inhibitor p16 (INK4a) in replicative senescence of normal human fibroblasts. Proc Natl Acad Sci U S A 93(24): 13742-7.
Bae DS, Cho SB, Kim YJ, Whang JD, Song SY, Park CS, Kim DS, Lee JH. (2001) Aberrant expression of cyclin D1 is associated with poor prognosis in early stage cervical cancer of the uterus. Gynecol Oncol 81(3): 341-7.
Bartek J, Staskova Z, Draetta G, Lukas J. (1993) Molecular pathology of the cell cycle in human cancer cells. Stem Cells 11 Suppl1: 51-8.
Bialkowska-Hobrzanska H, Bowles L, Bukala B, Joseph MG, Fletcher R, Razvi H. (2000) Comparison of human telomerase reverse transcriptase messenger RNA and telomerase activity as urine markers for diagnosis of bladder carcinoma. Mol Diagn 5(4): 267-77.
Bieche I, Nogues C, Paradis V, Olivi M, Bedossa P, Lidereau R, Vidaud M. (2000) Quantitation of hTERT gene expression in sporadic breast tumors with a real-time reverse transcription-polymerase chain reaction assay. Clin Cancer Res 6(2): 452-9.
Blackburn EH. (1990) Telomeres: structure and synthesis. J Biol Chem 265(11): 5919-21.
Blackburn EH. (1991) Structure and function of telomeres. Nature 350(6319): 569-73.
Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE. (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279(5349): 349-52.
Breitschopf K, Zeiher AM, Dimmeler S. (2001) Pro-atherogenic factors induce telomerase inactivation in endothelial cells through an Akt-dependent mechanism. FEBS Lett 493(1): 21-5.
Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, and James D. Watson. Published by Garland Publishing, a member of the Taylor & Francis Group. (1994) Basic Genetic Mechanisms: DNA Replication. Molecular Biology of the Cell Part II. Molecular Genetics Chapter 6.
Bruce JL, Hurford RK Jr, Classon M, Koh J, Dyson N. (2000) Requirements for cell cycle arrest by p16INK4a. Mol Cell 6(3): 737-42.
Buys CH. (2000) Telomeres, telomerase, and cancer. N Engl J Med 342(17): 1282-3
Cairns P, Tokino K, Eby Y, Sidransky D. (1994) Homozygous deletions of 9p21 in primary human bladder tumors detected by comparative multiplex polymerase chain reaction. Cancer Res 54(6): 1422-4.
Castellano M, Pollock PM, Walters MK, Sparrow LE, Down LM, Gabrielli BG, Parsons PG, Hayward NK.(1997) CDKN2A/p16 is inactivated in most melanoma cell lines. Cancer Res 57(21): 4868-75.
Cerni C. (2000) Telomeres, telomerase, and myc. An update. Mutat Res 462(1): 31-47.
Cerone MA, Londono-Vallejo JA, Bacchetti S. (2001) Telomere maintenance by telomerase and by recombination can coexist in human cells. Hum Mol Genet 10(18): 1945-52.
Connell-Crowley L, Harper JW, Goodrich DW. (1997) Cyclin D1/Cdk4 regulates retinoblastoma protein-mediated cell cycle arrest by site-specific phosphorylation. Mol Biol Cell 8(2): 287-301.
Counter CM, Hahn WC, Wei W, Caddle SD, Beijersbergen RL, Lansdorp PM, Sedivy JM, Weinberg RA. (1998) Dissociation among in vitro telomerase activity, telomere maintenance, and cellular immortalization. Proc Natl Acad Sci U S A 95(25): 14723-8.
Dahse R, Fiedler W, Ernst G. (1997) Telomeres and telomerase. Pathologe 18(6): 425-9.
Dahse R, Mey J. (2001) Telomerase in human tumors: molecular diagnosis and clinical significance. Expert Rev Mol Diagn 1 (2): 201-10.
Dang CV. (1999) c-Myc target genes involved in cell growth, apoptosis, and metabolism. Mol Cell Biol 19(1): 1-11
De Braud F, Maffezzini M, Vitale V, Bruzzi P, Gatta G, Hendry WF, Sternberg CN. (2002) Bladder cancer. Crit Rev Oncol Hematol 41(1): 89-106.
De Kok JB, Schalken JA, Aalders TW, Ruers TJ, Willems HL, Swinkels DW. (2000) Quantitative measurement of telomerase reverse transcriptase (hTERT) mRNA in urothelial cell carcinomas. Int J Cancer 87(2): 217-20.
Dolin PJ, Cook-Mozaffari P. (1992) Occupation and bladder cancer: a death-certificate study. Br J Cancer 66(3): 568-78.
Donato F, Boffetta P, Fazioli R, Aulenti V, Gelatti U, Porru S. (1997) Bladder cancer, tobacco smoking, coffee and alcohol drinking in Brescia, northern Italy. Eur J Epidemiol 13(7): 795-800.
Drissi R, Zindy F, Roussel MF, Cleveland JL. (2001) c-Myc-mediated regulation of telomerase activity is disabled in immortalized cells. J Biol Chem 276(32): 29994-30001.
Ducrest AL, Amacker M, Mathieu YD, Cuthbert AP, Trott DA, Newbold RF, Nabholz M, Lingner J.(2001) Regulation of human telomerase activity: repression by normal chromosome 3 abolishes nuclear telomerase reverse transcriptase transcripts but does not affect c-Myc activity. Cancer Res 61(20): 7594-602.
Duncan EL, Wadhwa R, Kaul SC. (2000) Senescence and immortalization of human cells. Biogerontology 1(2): 103-21.
Dyson N. (1998) The regulation of E2F by pRB-family proteins. Genes Dev 12(15): 2245-62.
Edwards J, Duncan P, Going JJ, Grigor KM, Watters AD, Bartlett JM. (2001) Loss of heterozygosity on chromosomes 11 and 17 are markers of recurrence in TCC of the bladder. Br J Cancer 85(12): 1894-9.
Eissa S, Ali-Labib R, Khalifa A. (2000) Deletion of p16 and p15 genes In schistosomiasis-associated bladder cancer (SABC). Clin Chim Acta 300(1-2): 159-69.
Fan Z, Zhang J, Bai J. (2001) Deletion mapping of chromosome 18q and Smad proteins expression analysis in urinary bladder carcinomas. Zhonghua Zhong Liu Za Zhi 23(2): 135-7.
Feng J, Funk WD, Wang SS, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, et al. (1995) The RNA component of human telomerase. Science 269(5228): 1236-41.
Finnon P, Silver AR, Bouffler SD. (2000) Upregulation of telomerase activity by X-irradiation in mouse leukaemia cells is independent of Tert, Terc, Tnks and Myc transcription. Carcinogenesis 21(4): 573-8.
Foster JS, Henley DC, Bukovsky A, Seth P, Wimalasena J. (2001) Multifaceted regulation of cell cycle progression by estrogen: regulation of Cdk inhibitors and Cdc25A independent of cyclin D1-Cdk4 functionMol Cell Biol 21(3): 794-810.
Gazzaniga P, Gradilone A, Frati L, Agliano AM. (2001) Epidermal growth factor receptor mRNA expression in peripheral blood of bladder cancer patients: a potential marker to detect treatment failure. Clin Cancer Res7 (12): 4288-9.
Giroldi LA, Bringuier PP, Shimazui T, Jansen K, Schalken JA. (1999) Changes in cadherin-catenin complexes in the progression of human bladder carcinoma. Int J Cancer 82(1): 70-6.
Gonzalez-Quevedo R, Iniesta P, Moran A, de Juan C, Sanchez-Pernaute A, Fernandez C, Torres A, Diaz-Rubio E, Balibrea JL, Benito M. (2002) Cooperative role of telomerase activity and p16 expression in the prognosis of non-small-cell lung cancer. J Clin Oncol 20(1): 254-62.
Granger MP, Wright WE, Shay JW. (2002) Telomerase in cancer and aging. Crit Rev Oncol Hematol 41(1): 29-40.
Greenberg RA, O'Hagan RC, Deng H, Xiao Q, Hann SR, Adams RR, Lichtsteiner S, Chin L, Morin GB, DePinho RA. (1999) Telomerase reverse transcriptase gene is a direct target of c-Myc but is not functionally equivalent in cellular transformation. Oncogene 18(5): 1219-26.
Greider CW, Blackburn EH. (1989) A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature 337(6205): 331-7.
Greider CW. (1996) Telomere length regulation. Annu Rev Biochem 65: 337-65.
Haas K, Staller P, Geisen C, Bartek J, Eilers M, Moroy T. (1997) Mutual requirement of CDK4 and Myc in malignant transformation: evidence for cyclin D1/CDK4 and p16INK4A as upstream regulators of Myc. Oncogene 15(2): 179-92.
Hara E, Smith R, Parry D, Tahara H, Stone S, Peters G. (1996) Regulation of p16CDKN2 expression and its implications for cell immortalization and senescence. Mol Cell Biol 16(3): 859-67
Hara T, Noma T, Yamashiro Y, Naito K, Nakazawa A. (2001) Quantitative analysis of telomerase activity and telomerase reverse transcriptase expression in renal cell carcinoma. Urol Res 29(1): 1-6.
Harada K, Kurisu K, Tahara H, Tahara E, Ide T, Tahara E. (2000) Telomerase activity in primary and secondary glioblastomas multiforme as a novel molecular tumor marker J Neurosurg 93(4):618-25.
Harrington L, McPhail T, Mar V, Zhou W, Oulton R, Bass MB, Arruda I, Robinson MO. (1997) A mammalian telomerase-associated protein. Science 275(5302): 973-7.
Hashimoto K, Nakagawa Y, Morikawa H, Niki M, Egashira Y, Hirata I, Katsu K, Akao Y. (2001) Co-overexpression of DEAD box protein rck/p54 and c-myc protein in human colorectal adenomas and the relevance of their expression in cultured cell lines. Carcinogenesis; 22(12): 1965-70
Henson JD, Neumann AA, Yeager TR, Reddel RR. (2002) Alternative lengthening of telomeres in mammalian cells. Oncogene 21(4): 598-610.
Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ, O'Connell BC, Mateyak MK, Tam W, Kohlhuber F, Dang CV, Sedivy JM, Eick D, Vogelstein B, Kinzler KW. (2000) Identification of CDK4 as a target of c-MYC. Proc Natl Acad Sci U S A 97(5): 2229-34.
Himber J. (1993) Horizontal semi-dry electroblotting for the detection of the low density lipoprotein receptor in solubilized liver membranes. Electrophoresis 14(8): 794-7.
Hinds PW, Dowdy SF, Eaton EN, Arnold A, Weinberg RA. (1994) Function of a human cyclin gene as an oncogene. Proc. Nat. Acad. Science 91: 709-713.
Hiyama E, Hiyama K, Yokoyama T, Matsuura Y, Piatyszek MA, Shay JW. (1995) Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med 1(3): 249-55.
Hohaus S, Voso MT, Orta-La Barbera E, Cavallo S, Bellacosa A, Rutella S, Rumi C, Genuardi M, Neri G, Leone G. (1997) Telomerase activity in human hematopoietic progenitor cells. Haematologica 82(3): 262-8.
Hoshi S, Shintaku I, Suzuki K, Takahashi T, Kaihou Y, Ishidoya S, Namima T, Ohyama C, Orikasa S. (2000) Bladder preservation by internal iliac arterial infusion chemotherapy and irradiation in T3 bladder carcinoma patients over the age of 70 years. Tohoku J Exp Med192 (4): 249-58.
Howe GR, Burch JD, Miller AB, Cook GM, Esteve J, Morrison B, Gordon P, Chambers LW, Fodor G, Winsor GM. (1980) Tobacco use, occupation, coffee, various nutrients, and bladder cancer. J Natl Cancer Inst 64(4): 701-13.
Jares P, Nadal A, Fernandez PL, Pinyol M, Hernandez L, Cazorla M, Hernandez S, Bea S, Cardesa A, Campo E. (1999) Disregulation of p16MTS1/CDK4I protein and mRNA expression is associated with gene alterations in squamous-cell carcinoma of the larynx. Int J Cancer 81(5): 705-11.
Jin Y, Cerletti N. Western blotting of transforming growth factor beta 2. (1992) Optimization of the electrophoretic transfer. Appl Theor Electrophor 3(2): 85-90.
Jong HS, Park YI, Kim S, Sohn JH, Kang SH, Song SH, Bang YJ, Kim NK. (1999) Up-regulation of human telomerase catalytic subunit during gastric carcinogenesis. Cancer 86(4): 559-65.
Joo M, Kang YK, Kim MR, Lee HK, Jang JJ. (2001) Cyclin D1 overexpression in hepatocellular carcinoma. Liver 21(2): 89-95.
Jung I, Messing E. (2000) Molecular mechanisms and pathways in bladder cancer development and progression. Cancer Control 7(4): 325-34.
Kamb A, Shattuck-Eidens D, Eeles R, Liu Q, Gruis NA, Ding W, Hussey C, Tran T, Miki Y, Weaver-Feldhaus J, et al. (1994) Analysis of the p16 gene (CDKN2) as a candidate for the chromosome 9p melanoma susceptibility locus. Nature Genet. 8: 22-26.
Kamb, A.; Gruis, N. A.; Weaver-Feldhaus, J.; Liu, Q.; Harshman, K.; Tavtigian, S. V.; Stockert, E.; Day, R. S., III; Johnson, B. E.; Skolnick, M. H. (1994) A cell cycle regulator potentially involved in genesis of many tumor types. Science 264: 436-440.
Kass-Eisler A, Greider CW. (2000) Recombination in telomere-length maintenance. Trends Biochem Sci 25 (4): 200-4.
Kato JY, Sherr CJ. (1993) Inhibition of granulocyte differentiation by G1 cyclins D2 and D3 but not D1. Proc Natl Acad Sci U S A 90(24): 11513-7.
Kim H, You S, Farris J, Kong BW, Christman SA, Foster LK, Foster DN. (2002) Expression profiles of p53-, p16(INK4a)-, and telomere-regulating genes in replicative senescent primary human, mouse, and chicken fibroblast cells. Exp Cell Res 272 (2): 199-208.
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL, Shay JW. (1994) Specific association of human telomerase activity with immortal cells and cancer. Science 266(5193): 2011-5.
Kimura F, Florl AR, Seifert HH, Louhelainen J, Maas S, Knowles MA, Schulz WA. (2001) Destabilization of chromosome 9 in transitional cell carcinoma of the urinary bladder. Br J Cancer 85(12): 1887-93.
Klobutcher LA, Swanton MT, Donini P, Prescott DM. ( 1981 ) All gene-sized DNA molecules in four species of hypotrichs have the same terminal sequence and an unusual 3' terminus. Proc Natl Acad Sci U S A 78(5): 3015-9
Kolquist, K. A.; Ellisen, L. W.; Counter, C. M.; Meyerson, M.; Tan, L. K.; Weinberg, R. A.; Haber, D. A.; Gerald, W. L.(1998) Expression of TERT in early premalignant lesions and a subset of cells in normal tissues. Nature Genet. 19: 182-186.
Krams M, Claviez A, Heidorn K, Krupp G, Parwaresch R, Harms D, Rudolph P. (2001) Regulation of telomerase activity by alternate splicing of human telomerase reverse transcriptase mRNA in a subset of neuroblastomas Am J Pathol 159(5): 1925-32.
Krupp G, Bonatz G, Parwaresch R. (2000) Telomerase, immortality and cancer. Biotechnol Annu Rev 6: 103-40.
Kyo S, Takakura M, Taira T, Kanaya T, Itoh H, Yutsudo M, Ariga H, Inoue M. (2000) Sp1 cooperates with c-Myc to activate transcription of the human telomerase reverse transcriptase gene (hTERT). Nucleic Acids Res 28(3): 669-77.
Kyo S, Takakura M, Tanaka M, Kanaya T, Sagawa T, Kohama T, Ishikawa H, Nakano T, Shimoya K, Inoue M. (1997) Expression of telomerase activity in human chorion. Biochem Biophys Res Commun 241(2): 498-503.
Lai S, El-Naggar AK. (1999) Differential expression of key cell cycle genes (p16/cyclin D1/pRb) in head and neck squamous carcinomas. : Lab Invest 79(3): 255-60.
Langford LA, Piatyszek MA, Xu R, Schold SC Jr, Wright WE, Shay JW. (1997) Telomerase activity in ordinary meningiomas predicts poor outcome. Hum Pathol 28(4): 416-20.
Latil A, Vidaud D, Valeri A, Fournier G, Vidaud M, Lidereau R, Cussenot O, Biache I. (2000) htert expression correlates with MYC over-expression in human prostate cancer. Int J Cancer 89(2): 172-6.
Leber B, Bacchetti S. (1996) Telomeres and telomerase in normal and malignant haematologic cells. Leuk Lymphoma 24(1-2):1-9.
Lee CT, Seol JY, Park KH, Yoo CG, Kim YW, Ahn C, Song YW, Han SK, Han JS, Kim S, Lee JS, Shim YS. (2001) Differential effects of adenovirus-p16 on bladder cancer cell lines can be overcome by the addition of butyrate. Clin Cancer Res 7(1): 210-4.
Liao J, Xu X, Wargovich MJ. (2000) Direct reprobing with anti-beta-actin antibody as an internal control for western blotting analysis. Biotechniques 28(2): 216-8.
Lipponen PK. (1995) Expression of c-myc protein is related to cell proliferation and expression of growth factor receptors in transitional cell bladder cancerJ Pathol 175(2): 203-10.
Liu BC, Loughlin KR. (2000) Telomerase in human bladder cancer. Urol Clin North Am 27(1): 115-23.
Liu JP. (1999) Studies of the molecular mechanisms in the regulation of telomerase activity. FASEB J 13(15): 2091-104.
Loughran O, Malliri A, Owens D, Gallimore PH, Stanley MA, Ozanne B, Frame MC, Parkinson EK. (1996) Association of CDKN2A/p16INK4A with human head and neck keratinocyte replicative senescence: relationship of dysfunction to immortality and neoplasia Oncogene 13(3): 561-8.
Lukas J, Aagaard L, Strauss M, Bartek J. (1995) Oncogenic aberrations of p16INK4/CDKN2 and cyclin D1 cooperate to deregulate G1 control. Cancer Res 55(21): 4818-23.
Lukas J, Pagano M, Staskova Z, Draetta G, Bartek J. (1994) Cyclin D1 protein oscillates and is essential for cell cycle progression in human tumour cell lines. Oncogene 9(3):707-18.
MacVicar AD. (2000) Bladder cancer staging. BJU Int 86 Suppl 1:111-22.
Mano Y, Shimizu T, Tanuma S, Takeda K. (2000) Synergistic down-regulation of telomerase activity and hTERT mRNA expression by combination of retinoic acid and GM-CSF in human myeloblastic leukemia ML-1 cells. Anticancer Res20 (3A): 1649-52.
Mayfield MP, Shah T, Flannigan GM, Hamilton Stewart PA, Bibby MC. (1998) Telomerase activity in malignant and benign bladder conditions. Int J Mol Med 1(5): 835-40.
McClintock B. (1938) The production of homozygous deficient tissues with mutant characteristics by means of the aberrant behavior of ring-shaped chromosomes. Genetics 23: 215-376.
Meyerson, M.; Counter, C. M.; Eaton, E. N.; Ellisen, L. W.; Steiner, P.; Caddle, S. D.; Ziaugra, L.; Beijersbergen, R. L.; Davidoff, M. J.; Liu, Q.; Bacchetti, S.; Haber, D. A.; Weinberg, R. A. (1997) hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 90: 785-795.
Milde-Langosch K, Bamberger AM, Rieck G, Kelp B, Loning T. (2001) Overexpression of the p16 cell cycle inhibitor in breast cancer is associated with a more malignant phenotype. Breast Cancer Res Treat 67(1): 61-70.
Mirvish SS.(1995) Role of N-nitroso compounds (NOC) and N-nitrosation in etiology of gastric, esophageal, nasopharyngeal and bladder cancer and contribution to cancer of known exposures to NOC. Cancer Lett 93 (1): 17-48.
Miyao N, Tsai YC, Lerner SP, Olumi AF, Spruck CH 3rd, Gonzalez-Zulueta M, Nichols PW, Skinner DG, Jones PA. (1993) Role of chromosome 9 in human bladder cancer. Cancer Res 53(17): 4066-70.
Morin, GB. (1989) The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell 59: 521-529.
Muller H, Lukas J, Schneider A, Warthoe P, Bartek j, Eilers M,
Strauss M. (1994) Cyclin D1 expression is regulated by the retinoblastoma protein. Proc.Nat.Acad. Science 91: 2945-2949.
Muller HJ. (1938) The remaking of chromosomes. Collecting Net 8:182-195.
Muller M, Heicappell R, Krause H, Sachsinger J, Porsche C, Miller K. (1999) Telomerase activity in malignant and benign renal tumors. Eur Urol 35(3): 249-55.
Muller M. (2002) Telomerase: its clinical relevance in the diagnosis of bladder cancer. Oncogene 21(4): 650-5.
Nakano K, Watney E, McDougall JK. (1998) Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer. Am J Pathol 153(3): 857-64.
Navon JD, Soliman H, Khonsari F, Ahlering T. (1997) Screening cystoscopy and survival of spinal cord injured patients with squamous cell cancer of the bladder. J Urol 157(6): 2109-11.
Neugut AI, Ahsan H, Robinson E, Ennis RD. (1997) Bladder carcinoma and other second malignancies after radiotherapy for prostate carcinoma. Cancer 79(8): 1600-4.
Nicol Keith W, Jeffry Evans TR, Glasspool RM. (2001) Telomerase and cancer: time to move from a promising target to a clinical reality. J Pathol 195(4): 404-14
Nishikawa R, Furnari FB, Lin H, Arap W, Berger MS, Cavenee WK, Su Huang HJ. (1995) Loss of P16INK4 expression is frequent in high grade gliomas. Cancer Res 55(9): 1941-5
Odom LD, Barrett JM, Pantazis CG, Stoddard LD, McDonough PG. (1989) Immunocytochemical study of ras and myc proto-oncogene polypeptide expression in the human menstrual cycle. Am J Obstet Gynecol 161(6 Pt 1): 1663-8.
Okamoto A, Demetrick DJ, Spillare EA, Hagiwara K, Hussain SP, Bennett WP, Forrester K, Gerwin B, Serrano M, Beach DH, et al. (1994) Mutations and altered expression of p16INK4 in human cancer. Proc Natl Acad Sci U S A 91(23):11045-9
Opitz OG, Suliman Y, Hahn WC, Harada H, Blum HE, Rustgi AK. (2001) Cyclin D1 overexpression and p53 inactivation immortalize primary oral keratinocytes by a telomerase-independent mechanism. J Clin Invest 108(5): 725-32.
Orlando C, Gelmini S, Selli C, Pazzagli M. (2001) Telomerase in urological malignancy. J Urol 166(2): 666-73.
Orlow I, LaRue H, Osman I, Lacombe L, Moore L, Rabbani F, Meyer F, Fradet Y, Cordon-Cardo C. (1999) Deletions of the INK4A gene in superficial bladder tumors. Association with recurrence. Am J Pathol 155(1): 105-13.
Orntoft TF, Wolf H. (1998) Molecular alterations in bladder cancer. Urol Res 26(4): 223-33.
Osman I, Scher H, Zhang ZF, Soos TJ, Hamza R, Eissa S, Khaled H, Koff A, Cordon-Cardo C. (1997) Expression of cyclin D1, but not cyclins E and A, is related to progression in bilharzial bladder cancer. Clin Cancer Res 3(12 Pt 1): 2247-51
Presti JC Jr, Reuter VE, Galan T, Fair WR, Cordon-Cardo C. (1991) Molecular genetic alterations in superficial and locally advanced human bladder cancer. Cancer Res 51(19): 5405-9.
Pusch O, Soucek T, Wawra E, Hengstschlager-Ottnad E, Bernaschek G, Hengstschlager M. (1996) Specific transformation abolishes cyclin D1 fluctuation throughout the cell cycle. FEBS Lett 385(3): 143-8.
Rabbani F, Cordon-Cardo C. (2000) Mutation of cell cycle regulators and their impact on superficial bladder cancer. Urol Clin North Am 27(1): 83-102.
Raymond S. Lance, Wadek. Aldons, Jason Blaser, J.Brantey Thrasher. (1998) Telomerase activity in solid transitional cell carcinoma, bladder washings, and voided urine. Urologic Oncology 4: 43-49.
Reddel RR. (1998) Genes involved in the control of cellular proliferative potential. Ann N Y Acad Sci 854: 8-19.
Reynisdottir I, Polyak K, Iavarone A, Massague J. (1995) Kip/Cip and Ink4 Cdk inhibitors cooperate to induce cell cycle arrest in response to TGF-beta. Genes Dev 9(15): 1831-45.
Reznikoff CA, Belair CD, Yeager TR, Savelieva E, Blelloch RH, Puthenveettil JA, Cuthill S. (1996) A molecular genetic model of human bladder cancer pathogenesis. Semin Oncol 23(5): 571-84.
Reznikoff CA, Sarkar S, Julicher KP, Burger MS, Puthenveettil JA, Jarrard DF, Newton MA. (2000) Genetic alterations and biological pathways in human bladder cancer pathogenesis. 1078-14395(5): 191-203
Rieger-Christ KM, Cain JW, Braasch JW, Dugan JM, Silverman ML, Bouyounes B, Libertino JA, Summerhayes IC. (2001) Expression of classic cadherins type I in urothelial neoplastic progression. Hum Pathol 32(1): 18-23.
Ripert C. (2000) Urinary schistosomiasis. Presse Med 29 (28):1573-6.
Rischmann P. (2002) Diagnosis of bladder tumors Rev Prat 52 (1):32-5.
Rohde V, Sattler HP, Bund T, Bonkhoff H, Fixemer T, Bachmann C, Lensch R, Unteregger G, Stoeckle M, Wullich B. (2000) Expression of the human telomerase reverse transcriptase is not related to telomerase activity in normal and malignant renal tissue. Clin Cancer Res 6(12): 4803-9.
Rosenberg, C. L.; Wong, E.; Petty, E. M.; Bale, A. E.; Tsujimoto, Y.; Harris, N. L.; Arnold, A. (1991) PRAD1, a candidate BCL1 oncogene: mapping and expression in centrocytic lymphoma. Proc. Nat. Acad. Sci. 88: 9638-9642.
Ross JS, Cohen MB. (2001) Detecting recurrent bladder cancer: new methods and biomarkers. Expert Rev Mol Diagn 1(1): 39-52.
Saad A, Hanbury DC, McNicholas TA, Boustead GB, Morgan S, Woodman AC. (2002) A study comparing various noninvasive methods of detecting bladder cancer in urine. BJU Int 89(4): 369-73.
Sagawa Y, Nishi H, Isaka K, Fujito A, Takayama M. (2001) The correlation of TERT expression with c-myc expression in cervical cancer. : Cancer Lett 168(1): 45-50.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning : A Laboratory Manual , second edition.
Saran KK, Gould D, Godec CJ, Verma RS. (1996) Genetics of bladder cancer. J Mol Med 74(8): 441-5.
Sauter G, Carroll P, Moch H, Kallioniemi A, Kerschmann R, Narayan P, Mihatsch MJ, Waldman FM. (1995) c-myc copy number gains in bladder cancer detected by fluorescence in situ hybridization. Am J Pathol 146(5): 1131-9.
Schmitz-Drager BJ, Schulz WA, Jurgens B, Gerharz CD, van Roeyen CR, Bultel H, Ebert T, Ackermann R. (1997) c-myc in bladder cancer. Clinical findings and analysis of mechanism. Urol Res 25 Suppl 1: S45-9.
Serrano M, Hannon GJ, Beach D. (1993) A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature 366(6456): 704-7.
Shay JW, Bacchetti S. (1997): A survey of telomerase activity in human cancer. Eur J Cancer 33(5): 787-91.
Shay JW, Wright WE. (1996) Telomerase activity in human cancer. Curr Opin Oncol 8(1): 66-71.
Sherr CJ. (1995) D-type cyclins. Trends Biochem Sci 20 (5):187-90.
Shin KY, Kong G, Kim WS, Lee TY, Woo YN, Lee JD. (1997) Overexpression of cyclin D1 correlates with early recurrence in superficial bladder cancers. Br J Cancer 75(12): 1788-92.
Simon M, Park T W, Koster G, Mahlberg R, Hackenbroch M, Bostrom J, Loning T, Schramm J. (2001) Alterations of INK4a (p16-p14ARF)/INK4b (p15) expression and telomerase activation in meningioma progression. J Neurooncol 55(3): 149-58.
Simon M, Park TW, Leuenroth S, Hans VH, Loning T, Schramm J. (2000) Telomerase activity and expression of the telomerase catalytic subunit, hTERT, in meningioma progression. J Neurosurg 92(5): 832- 40.
Steele EC Jr, Wang JH, Lo WK, Saperstein DA, Li X, Church RL. (2000) Lim2 (To3) transgenic mice establish a causative relationship between the mutation identified in the lim2 gene and cataractogenesis in the To3 mouse mutant. Mol Vis 6:85-94.
Stemmermann G, Heffelfinger SC, Noffsinger A, Hui YZ, Miller MA, Fenoglio-Preiser CM. (1994) The molecular biology of esophageal and gastric cancer and their precursors: oncogenes, tumor suppressor genes, and growth factors. Hum Pathol 25(10): 968-81.
Suenaga M, Soda H, Oka M, Yamaguchi A, Nakatomi K, Shiozawa K, Kawabata S, Kasai T, Yamada Y, Kamihira S, Tei C, Kohno S. (2002) Histone deacetylase inhibitors suppress telomerase reverse transcriptase mRNA expression in prostate cancer cells. Int J Cancer 97(5): 621-5.
Sugita Y, Nakashima A, Kato S, Sakata K, Morimatsu M, Shigemori M. (2000) Telomerase activity in gliomas with the use of non-radioisotopic and semi-quantitative procedure for terminal repeat amplification protocol. Oncol Rep7(5):1087-92.
Suwa Y, Takano Y, Iki M, Takeda M, Asakura T, Noguchi S, Masuda M. (1998) Cyclin D1 protein overexpression is related to tumor differentiation, but not to tumor progression or proliferative activity, in transitional cell carcinoma of the bladder. J Urol 160(3 Pt 1): 897-900.
Suzuki T, Suzuki Y, Fujioka T. (1999) Expression of the catalytic subunit associated with telomerase gene in human urinary bladder cancer. J Urol 162(6): 2217-20.
Sviderskaya EV, Hill SP, Evans-Whipp TJ, Chin L, Orlow SJ, Easty DJ, Cheong SC, Beach D, DePinho RA, Bennett DC. (2002) p16 (Ink4a) in melanocyte senescence and differentiation. J Natl Cancer Inst 94(6): 446-54.
Syrigos KN, Harrington K, Waxman J, Krausz T, Pignatelli M. (1998) Altered gamma-catenin expression correlates with poor survival in patients with bladder cancer. J Urol 160(5): 1889-93.
Takakura M, Kyo S, Kanaya T, Hirano H, Takeda J, Yutsudo M, Inoue M. (1999) Cloning of human telomerase catalytic subunit (hTERT) gene promoter and identification of proximal core promoter sequences essential for transcriptional activation in immortalized and cancer cells. Cancer Res 59(3): 551-7.
Takakura M, Kyo S, Sowa Y, Wang Z, Yatabe N, Maida Y, Tanaka M, Inoue M. (2001) Telomerase activation by histone deacetylase inhibitor in normal cells. Nucleic Acids Res 29(14): 3006-11.
Terasawa K, Sagae S, Takeda T, Ishioka S, Kobayashi K, Kudo R. (1999) Telomerase activity in malignant ovarian tumors with deregulation of cell cycle regulatory proteins. Cancer Lett 142(2): 207-17.
Thiery JP. (2001) Role of growth factor signaling in epithelial cell plasticity during development and in carcinogenesis. Bull Acad Natl Med 185(7): 1279-92; discussion 1293-4.
Torben F., Orntoft.Hans Wolf. (1998) Molecular alteration in bladder cancer. Urol Res 26: 223-233.
Tsujimoto, Y.; Yunis, J.; Onorato-Showe, L.; Erikson, J.; Nowell, P. C.; Croce, C. M. (1984) Molecular cloning of the chromosomal breakpoint of B-cell lymphomas and leukemias with the t(11;14) chromosome translocation. Science 224: 1403-1406.
Ueda M, Ouhtit A, Bito T, Nakazawa K, Lubbe J, Ichihashi M, Yamasaki H, Nakazawa H. (1997) Evidence for UV-associated activation of telomerase in human skin. Cancer Res 57(3): 370-4.
Ulaner GA, Hu JF, Vu TH, Giudice LC, Hoffman AR. (1998) Telomerase activity in human development is regulated by human telomerase reverse transcriptase (hTERT) transcription and by alternate splicing of hTERT transcripts. Cancer Res 58(18): 4168-72.
Ulaner GA, Hu JF, Vu TH, Giudice LC, Hoffman AR. (2001) Tissue-specific alternate splicing of human telomerase reverse transcriptase (hTERT) influences telomere lengths during human development. Int J Cancer 91(5): 644-9.
Vaziri H, Benchimol S. (1999) Alternative pathways for the extension of cellular life span: inactivation of p53/pRb and expression of telomerase. Oncogene 18(53): 7676-80.
Volm T, Pfaff P, Gnann R, Kreienberg R. (2001) Bladder carcinoma associated with cyclophosphamide therapy for ovarian cancer occurring with a latency of 20 years. Gynecol Oncol 82(1): 197-9.
Wainwright LJ, Lasorella A, Iavarone A. (2001) Distinct mechanisms of cell cycle arrest control the decision between differentiation and senescence in human neuroblastoma cells. Proc Natl Acad Sci U S A 98(16): 9396-400.
Wang J, Xie LY, Allan S, Beach D, Hannon GJ. (1998) Myc activates telomerase. Genes Dev 12(12): 1769-74.
Wang W, Wu J, Zhang Z, Tong T. (2001) Characterization of regulatory elements on the promoter region of p16(INK4a) that contribute to overexpression of p16 in senescent fibroblasts. J Biol Chem 276(52): 48655-61.
Warren JW. (1997) Catheter-associated urinary tract infections. : Infect Dis Clin North Am 11(3): 609-22.
Watson JD.(1972) Origin of concatemeric T7 DNA. Nat New Biol 239(94): 197-201.
Wilkie, A. O.; Lamb, J.; Harris, P. C.; Finney, R. D.; Higgs, D. R. (1990) A truncated human chromosome 16 associated with alpha thalassaemia is stabilized by addition of telomeric repeat (TTAGGG)n. Nature 346: 868-871.
Williams SG, Buscarini M, Stein JP. (2001) Molecular markers for diagnosis, staging, and prognosis of bladder cancer. Oncology (Huntingt) 15(11): 1461-70, 1473-4, 1476; discussion 1476-84.
Wine RN, Chapin RE. (1999) Adhesion and signaling proteins spatiotemporally associated with spermiation in the rat. J Androl 20(2): 198-213.
Wisman GB, Knol AJ, Helder MN, Krans M, de Vries EG, Hollema H, de Jong S, van der Zee AG. (2001): Telomerase in relation to clinicopathologic prognostic factors and survival in cervical cancer. Int J Cancer 91(5): 658-64.
Wright WE, Pereira-Smith OM, Shay JW. (1989) Reversible cellular senescence: implications for immortalization of normal human diploid fibroblasts. Mol Cell Biol 9(7): 3088-92.
Wu KJ, Grandori C, Amacker M, Simon-Vermot N, Polack A, Lingner J. (1999) Direct activation of TERT transcription by c-Myc. Nat Genet 21:220-224.
Wu WJ, Liu LT, Huang CN, Huang CH, Chang LL. (2000) The clinical implications of telomerase activity in upper tract urothelial cancer and washings. BJU Int 86(3): 213-9.
Wu WJ, Liu LT, Huang CN, Huang CH, Chang SF, Chang LL. (2001) Telomerase activity in human bladder tumors and bladder washing specimes. Kaohsing J Med Sci 17: 602-609.
Xu D, Popov N, Hou M, Wang Q, Bjorkholm M, Gruber A, Menkel AR, Henriksson M. (2001) Switch from Myc/Max to Mad1/Max binding and decrease in histone acetylation at the telomerase reverse transcriptase promoter during differentiation of HL60 cells. Proc Natl Acad Sci U S A 98(7): 3826-31.
Yang MH, Chen KK, Yen CC, Wang WS, Chang YH, Huang WJ, Fan FS, Chiou TJ, Liu JH, Chen PM. (2002) Unusually high incidence of upper urinary tract urothelial carcinoma in Taiwan. Urology 59(5): 681-7.
Yeager TR, Neumann AA, Englezou A, Huschtscha LI, Noble JR, Reddel RR. (1999) Telomerase-negative immortalized human cells contain a novel type of promyelocytic leukemia (PML) body. Cancer Res 59(17): 4175-9.
Yokota K, Kanda K, Inoue Y, Kanayama H, Kagawa S. (1998) Semi-quantitative analysis of telomerase activity in exfoliated human urothelial cells and bladder transitional cell carcinoma. Br J Urol 82(5): 727-32.
Yoo J, Park SY, Kang SJ, Shim SI, Kim BK. (2002) Altered expression of G1 regulatory proteins in human soft tissue sarcomas. Arch Pathol Lab Med 126(5): 567-73.