Cloning of the 5’Upstream Region of the Rat p16 Gene and Its Role in Silencing

Masanobu Abe; Eriko Okochi; Takashi Kuramoto; Atsushi Kaneda; Tsuyoshi Takato; Takashi Sugimura; Toshikazu Ushijima

doi:10.1111/j.1349-7006.2002.tb01211.x

. 2002 Oct;93(10):1100–1106. doi: 10.1111/j.1349-7006.2002.tb01211.x

Cloning of the 5’Upstream Region of the Rat p16 Gene and Its Role in Silencing

Masanobu Abe ^1,², Eriko Okochi ¹, Takashi Kuramoto ¹, Atsushi Kaneda ¹, Tsuyoshi Takato ², Takashi Sugimura ¹, Toshikazu Ushijima ^1,^✉

PMCID: PMC5926886 PMID: 12417039

Abstract

Hypermethylation of the 5’upstream region (5’region) of the human p16^CDKN2A (p16) gene is known to cause silencing, which is involved in a wide range of human cancers. For the rat p16 gene, its 5’region has not been cloned, and it is uncertain whether surrogate use of exon la is adequate for analysis of p16 silencing. In this study, we observed that methylation analysis of exon la gave false positive results in three samples of normal rat mammary epitheliums and in two of six primary mammary carcinomas. Therefore, we determined the nucleotide sequence of the 5’region of the rat p16 gene. To confirm that methylation status of the 5’region is correlated with p16 expression, the methylation status was analyzed by bisulfite sequencing and methylation‐specific PCR in three samples of normal mammary glands, six samples of mammary carcinomas and four cell lines. The 5’region was demethylated in all of the three normal and six carcinoma samples that fully expressed p16. On the other hand, the 5’region was highly methylated in the 3Y1 cell line, which lacked p16 expression, but without deletion. These results showed that the methylation status of the 5’region was more closely correlated with p16 expression than that of the exon 1α and analysis of the methylation status is useful in examining p16 silencing in various rat tumors.

Keywords: p16, Methylation, Rat, Silencing, CpG islands

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REFERENCES

1. ) Rocco , J. W. and Sidransky , D.p16 (MTS‐1/CDKN2/INK4a) in cancer progression . Exp. Cell Res. , 264 , 42 – 55 ( 2001. ). [DOI] [PubMed] [Google Scholar]
2. ) Liggett , W. H. , Jr. and Sidransky , D.Role of the p16 tumor suppressor gene in cancer . J. Clin. Oncol. , 16 , 1197 – 1206 ( 1998. ). [DOI] [PubMed] [Google Scholar]
3. ) Gonzalez‐Zulueta , M. , Bender , C. M. , Yang , A. S. , Nguyen , T. , Beart , R. W. , Van Tornout , J. M. and Jones , P. A.Methylation of the 5′ CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing . Cancer Res. , 55 , 4531 – 4535 ( 1995. ). [PubMed] [Google Scholar]
4. ) Merlo , A. , Herman , J. G. , Mao , L. , Lee , D. J. , Gabrielson , E. , Burger , P. C. , Baylin , S. B. and Sidransky , D.5′ CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers . Nat. Med. , 1 , 686 – 692 ( 1995. ). [DOI] [PubMed] [Google Scholar]
5. ) Gonzalgo , M. L. , Hayashida , T. , Bender , C. M. , Pao , M. M. , Tsai , Y. C. , Gonzales , F. A. , Nguyen , H. D. , Nguyen , T. T. and Jones , P. A.The role of DNA methylation in expression of the p19/p16 locus in human bladder cancer cell lines . Cancer Res. , 58 , 1245 – 1252 ( 1998. ). [PubMed] [Google Scholar]
6. ) Baylin , S. B. and Herman , J. G.DNA hypermethylation in tumorigenesis: epigenetics joins genetics . Trends Genet. , 16 , 168 – 174 ( 2000. ). [DOI] [PubMed] [Google Scholar]
7. ) Chan , M. F. , Liang , G. and Jones , P. A.Relationship between transcription and DNA methylation . Curr. Top. Microbiol. Immunol. , 249 , 75 – 86 ( 2000. ). [DOI] [PubMed] [Google Scholar]
8. ) Pilot , H. C. , Hikita , H. , Dragan , Y. , Sargent , L. and Haas , M.Review article: the stages of gastrointestinal carcinogenesis—application of rodent models to human disease . Aliment. Pharmacol. Ther. , 14 ( Suppl. 1 ), 153 – 160 ( 2000. ). [DOI] [PubMed] [Google Scholar]
9. ) Shirai , T. , Takahashi , S. , Cui , L. , Futakuchi , M. , Kato , K. , Tamano , S. and Imaida , K.Experimental prostate carcinogenesis—rodent models . Mutat. Res. , 462 , 219 – 226 ( 2000. ). [DOI] [PubMed] [Google Scholar]
10. ) Cohen , S. M.Alternative models for carcinogenicity testing: weight of evidence evaluations across models . Toxicol. Pathol. , 29 ( Suppl. ), 183 – 190 ( 2001. ). [DOI] [PubMed] [Google Scholar]
11. ) Stott , F. J. , Bates , S. , James , M. C. , McConnell , B. B. , Starborg , M. , Brookes , S. , Palmero , I. , Ryan , K. , Kara , E. , Vousden , K. H. and Peters , G.The alternative product from the human CDKN2A locus, p14^ARF, participates in a regulatory feedback loop with p53 and MDM2 . EMBO J. , 17 , 5001 – 5014 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
12. ) Swafford , D. S. , Middleton , S. K. , Palmisano , W. A. , Nikula , K. J. , Tesfaigzi , J. , Baylin , S. B. , Herman , J. G. and Belinsky , S. A.Frequent aberrant methylation of p16^INK4a in primary rat lung tumors . Mol. Cell. Biol. , 17 , 1366 – 1374 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
13. ) Belinsky , S. A. , Nikula , K. J. , Palmisano , W. A. , Michels , R. , Saccomanno , G. , Gabrielson , E. , Baylin , S. B. and Herman , J. G.Aberrant methylation of p16^INK4ais an early event in lung cancer and a potential biomarker for early diagnosis . Proc. Natl. Acad. Set. USA , 95 , 11891 – 11896 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
14. ) Tanaka , T. , Iwasa , Y. , Kondo , S. , Hiai , H. and Toyokuni , S.High incidence of allelic loss on chromosome 5 and inactivation of p15^INK4B and p16^INK4A tumor suppressor genes in oxystress‐induced renal cell carcinoma of rats . Oncogene , 18 , 3793 – 3797 ( 1999. ). [DOI] [PubMed] [Google Scholar]
15. ) Park , T. J. , Kim , H. S. , Byun , K. H. , Jang , J. J. , Lee , Y. S. and Lim , I. K.Sequential changes in hepatocarcinogenesis induced by diethylnitrosamine plus thioacetamide in Fischer 344 rats: induction of gankyrin expression in liver fibrosis, pRB degradation in cirrhosis, and methylation of p16^INK4A exon 1 in hepatocellular carcinoma . Mol. Carcinog. , 30 , 138 – 150 ( 2001. ). [DOI] [PubMed] [Google Scholar]
16. ) Pulling , L. C. , Klinge , D. M. and Belinsky , S. A.P16^INK4lL and β‐catenin alterations in rat liver tumors induced by NNK . Cardnogenesis , 22 , 461 – 466 ( 2001. ). [DOI] [PubMed] [Google Scholar]
17. ) Belinsky , S. A. , Snow , S. S. , Nikula , K. J. , Finch , G. L. , Tellez , C. S. and Palmisano , W. A.Aberrant CpG island methylation of the p16^INK4a and estrogen receptor genes in rat lung tumors induced by particulate carcinogens . Cardnogenesis , 23 , 335 – 339 ( 2002. ). [DOI] [PubMed] [Google Scholar]
18. ) Watanabe , N. , Okochi , E. , Hirayama , Y. , Shimada , Y. , Yanagihara , K. , Yoshida , M. C. , Takahashi , S. , Mochizuki , M. , Sugimura , T. , Nagao , M. and Ushijima , T.Single nucleotide instability without microsatellite instability in rat mammary carcinomas . Cancer Res. , 61 , 2632 – 2640 ( 2001. ). [PubMed] [Google Scholar]
19. ) Okochi , E. , Watanabe , N. , Shimada , Y. , Takahashi , S. , Wakazono , K. , Shirai , T. , Sugimura , T. , Nagao , M. and Ushijima , T.Preferential induction of guanine deletion at 5′‐GGGA‐3′ in rat mammary glands by 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine . Cardnogenesis , 20 , 1933 – 1938 ( 1999. ). [DOI] [PubMed] [Google Scholar]
20. ) Sambrook , J. , Fritsch , E. F. and Maniatis , T.Isolation of high‐molecular‐weight DNA from mammalian cells . In “ Molecular Cloning ,” ed. Sambrook J. , Fritsch E. F. and Maniatis T. , pp. 9.14 – 19.23 ( 1989. ). Cold Spring Harbor Laboratory Press; , New York . [Google Scholar]
21. ) Kuramoto , T. , Kitada , K. , Inui , T. , Sasaki , Y. , Ito , K. , Hase , T. , Kawagachi , S. , Ogawa , Y. , Nakao , K. , Barsh , G. S. , Nagao , M. , Ushijima , T. and Serikawa , T.Attradin/ mahogany/zitter plays a critical role in myelination of the central nervous system . Proc. Natl. Acad. Sci. USA , 98 , 559 – 564 ( 2001. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
22. ) Rein , T. , Zorbas , H. and DePamphilis , M. L.Active mammalian replication origins are associated with a high‐density cluster of mCpG dinucleotides . Mol. Cell. Biol. , 17 , 416 – 426 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
23. ) Herman , J. G. , Graff , J. R. , Myohanen , S. , Nelkin , B. D. and Baylin , S. B.Methylation‐specific PCR: a novel PCR assay for methylation status of CpG islands . Proc. Natl. Acad. Sci. USA , 93 , 9821 – 9826 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
24. ) Gardiner‐Garden , M. and Frommer , M.CpG islands in vertebrate genomes . J. Mol. Biol. , 196 , 261 – 282 ( 1987. ). [DOI] [PubMed] [Google Scholar]
25. ) Issa , J. P. , Ottaviano , Y. L. , Celano , P. , Hamilton , S. R. , Davidson , N. E. and Baylin , S. B.Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon . Nat. Genet , 7 , 536 – 540 ( 1994. ). [DOI] [PubMed] [Google Scholar]
26. ) Miyakura , Y. , Sugano , K. , Konishi , F. , Ichikawa , A. , Maekawa , M. , Shitoh , K. , Igarashi , S. , Kotake , K. , Koyama , Y. and Nagai , H.Extensive methylation of hMLH1 promoter region predominates in proximal colon cancer with microsatellite instability . Gastroenterology , 121 , 1300 – 1309 ( 2001. ). [DOI] [PubMed] [Google Scholar]
27. ) Ushijima , T. , Makino , H. , Nakayasu , M. , Aonuma , S. , Takeuchi , M. , Segawa , K. , Sugimura , T. and Nagao , M.Presence of p53 mutations in 3Y1‐B clone 1–6: a rat cell line widely used as a normal immortalized fibroblast . Jpn. J. Cancer Res. , 85 , 455 – 458 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
28. ) Huschtscha , L. I. and Reddel , R. R.p16^INIK4a and the control of cellular proliferative life span . Cardnogenesis , 20 , 921 – 926 ( 1999. ). [DOI] [PubMed] [Google Scholar]
29. ) Wong , D. J. , Foster , S. A. , Galloway , D. A. and Reid , B. J.Progressive region‐specific de novo methylation of the p16 CpG island in primary human mammary epithelial cell strains during escape from M(0) growth arrest . Mol. Cell. Biol. , 19 , 5642 – 5651 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
30. ) Hui , R. , Macmillan , R. D. , Kenny , F. S. , Musgrove , E. A. , Blarney , R. W. , Nicholson , R. I. , Robertson , J. F. and Sutherland , R. L.INK4a gene expression and methylation in primary breast cancer: overexpression of p16^INK4a messenger RNA is a marker of poor prognosis . Clin. Cancer Res. , 6 , 2777 – 2787 ( 2000. ). [PubMed] [Google Scholar]

[b1] 1. ) Rocco , J. W. and Sidransky , D.p16 (MTS‐1/CDKN2/INK4a) in cancer progression . Exp. Cell Res. , 264 , 42 – 55 ( 2001. ). [DOI] [PubMed] [Google Scholar]

[b2] 2. ) Liggett , W. H. , Jr. and Sidransky , D.Role of the p16 tumor suppressor gene in cancer . J. Clin. Oncol. , 16 , 1197 – 1206 ( 1998. ). [DOI] [PubMed] [Google Scholar]

[b3] 3. ) Gonzalez‐Zulueta , M. , Bender , C. M. , Yang , A. S. , Nguyen , T. , Beart , R. W. , Van Tornout , J. M. and Jones , P. A.Methylation of the 5′ CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing . Cancer Res. , 55 , 4531 – 4535 ( 1995. ). [PubMed] [Google Scholar]

[b4] 4. ) Merlo , A. , Herman , J. G. , Mao , L. , Lee , D. J. , Gabrielson , E. , Burger , P. C. , Baylin , S. B. and Sidransky , D.5′ CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers . Nat. Med. , 1 , 686 – 692 ( 1995. ). [DOI] [PubMed] [Google Scholar]

[b5] 5. ) Gonzalgo , M. L. , Hayashida , T. , Bender , C. M. , Pao , M. M. , Tsai , Y. C. , Gonzales , F. A. , Nguyen , H. D. , Nguyen , T. T. and Jones , P. A.The role of DNA methylation in expression of the p19/p16 locus in human bladder cancer cell lines . Cancer Res. , 58 , 1245 – 1252 ( 1998. ). [PubMed] [Google Scholar]

[b6] 6. ) Baylin , S. B. and Herman , J. G.DNA hypermethylation in tumorigenesis: epigenetics joins genetics . Trends Genet. , 16 , 168 – 174 ( 2000. ). [DOI] [PubMed] [Google Scholar]

[b7] 7. ) Chan , M. F. , Liang , G. and Jones , P. A.Relationship between transcription and DNA methylation . Curr. Top. Microbiol. Immunol. , 249 , 75 – 86 ( 2000. ). [DOI] [PubMed] [Google Scholar]

[b8] 8. ) Pilot , H. C. , Hikita , H. , Dragan , Y. , Sargent , L. and Haas , M.Review article: the stages of gastrointestinal carcinogenesis—application of rodent models to human disease . Aliment. Pharmacol. Ther. , 14 ( Suppl. 1 ), 153 – 160 ( 2000. ). [DOI] [PubMed] [Google Scholar]

[b9] 9. ) Shirai , T. , Takahashi , S. , Cui , L. , Futakuchi , M. , Kato , K. , Tamano , S. and Imaida , K.Experimental prostate carcinogenesis—rodent models . Mutat. Res. , 462 , 219 – 226 ( 2000. ). [DOI] [PubMed] [Google Scholar]

[b10] 10. ) Cohen , S. M.Alternative models for carcinogenicity testing: weight of evidence evaluations across models . Toxicol. Pathol. , 29 ( Suppl. ), 183 – 190 ( 2001. ). [DOI] [PubMed] [Google Scholar]

[b11] 11. ) Stott , F. J. , Bates , S. , James , M. C. , McConnell , B. B. , Starborg , M. , Brookes , S. , Palmero , I. , Ryan , K. , Kara , E. , Vousden , K. H. and Peters , G.The alternative product from the human CDKN2A locus, p14^ARF, participates in a regulatory feedback loop with p53 and MDM2 . EMBO J. , 17 , 5001 – 5014 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b12] 12. ) Swafford , D. S. , Middleton , S. K. , Palmisano , W. A. , Nikula , K. J. , Tesfaigzi , J. , Baylin , S. B. , Herman , J. G. and Belinsky , S. A.Frequent aberrant methylation of p16^INK4a in primary rat lung tumors . Mol. Cell. Biol. , 17 , 1366 – 1374 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b13] 13. ) Belinsky , S. A. , Nikula , K. J. , Palmisano , W. A. , Michels , R. , Saccomanno , G. , Gabrielson , E. , Baylin , S. B. and Herman , J. G.Aberrant methylation of p16^INK4ais an early event in lung cancer and a potential biomarker for early diagnosis . Proc. Natl. Acad. Set. USA , 95 , 11891 – 11896 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b14] 14. ) Tanaka , T. , Iwasa , Y. , Kondo , S. , Hiai , H. and Toyokuni , S.High incidence of allelic loss on chromosome 5 and inactivation of p15^INK4B and p16^INK4A tumor suppressor genes in oxystress‐induced renal cell carcinoma of rats . Oncogene , 18 , 3793 – 3797 ( 1999. ). [DOI] [PubMed] [Google Scholar]

[b15] 15. ) Park , T. J. , Kim , H. S. , Byun , K. H. , Jang , J. J. , Lee , Y. S. and Lim , I. K.Sequential changes in hepatocarcinogenesis induced by diethylnitrosamine plus thioacetamide in Fischer 344 rats: induction of gankyrin expression in liver fibrosis, pRB degradation in cirrhosis, and methylation of p16^INK4A exon 1 in hepatocellular carcinoma . Mol. Carcinog. , 30 , 138 – 150 ( 2001. ). [DOI] [PubMed] [Google Scholar]

[b16] 16. ) Pulling , L. C. , Klinge , D. M. and Belinsky , S. A.P16^INK4lL and β‐catenin alterations in rat liver tumors induced by NNK . Cardnogenesis , 22 , 461 – 466 ( 2001. ). [DOI] [PubMed] [Google Scholar]

[b17] 17. ) Belinsky , S. A. , Snow , S. S. , Nikula , K. J. , Finch , G. L. , Tellez , C. S. and Palmisano , W. A.Aberrant CpG island methylation of the p16^INK4a and estrogen receptor genes in rat lung tumors induced by particulate carcinogens . Cardnogenesis , 23 , 335 – 339 ( 2002. ). [DOI] [PubMed] [Google Scholar]

[b18] 18. ) Watanabe , N. , Okochi , E. , Hirayama , Y. , Shimada , Y. , Yanagihara , K. , Yoshida , M. C. , Takahashi , S. , Mochizuki , M. , Sugimura , T. , Nagao , M. and Ushijima , T.Single nucleotide instability without microsatellite instability in rat mammary carcinomas . Cancer Res. , 61 , 2632 – 2640 ( 2001. ). [PubMed] [Google Scholar]

[b19] 19. ) Okochi , E. , Watanabe , N. , Shimada , Y. , Takahashi , S. , Wakazono , K. , Shirai , T. , Sugimura , T. , Nagao , M. and Ushijima , T.Preferential induction of guanine deletion at 5′‐GGGA‐3′ in rat mammary glands by 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine . Cardnogenesis , 20 , 1933 – 1938 ( 1999. ). [DOI] [PubMed] [Google Scholar]

[b20] 20. ) Sambrook , J. , Fritsch , E. F. and Maniatis , T.Isolation of high‐molecular‐weight DNA from mammalian cells . In “ Molecular Cloning ,” ed. Sambrook J. , Fritsch E. F. and Maniatis T. , pp. 9.14 – 19.23 ( 1989. ). Cold Spring Harbor Laboratory Press; , New York . [Google Scholar]

[b21] 21. ) Kuramoto , T. , Kitada , K. , Inui , T. , Sasaki , Y. , Ito , K. , Hase , T. , Kawagachi , S. , Ogawa , Y. , Nakao , K. , Barsh , G. S. , Nagao , M. , Ushijima , T. and Serikawa , T.Attradin/ mahogany/zitter plays a critical role in myelination of the central nervous system . Proc. Natl. Acad. Sci. USA , 98 , 559 – 564 ( 2001. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b22] 22. ) Rein , T. , Zorbas , H. and DePamphilis , M. L.Active mammalian replication origins are associated with a high‐density cluster of mCpG dinucleotides . Mol. Cell. Biol. , 17 , 416 – 426 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b23] 23. ) Herman , J. G. , Graff , J. R. , Myohanen , S. , Nelkin , B. D. and Baylin , S. B.Methylation‐specific PCR: a novel PCR assay for methylation status of CpG islands . Proc. Natl. Acad. Sci. USA , 93 , 9821 – 9826 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b24] 24. ) Gardiner‐Garden , M. and Frommer , M.CpG islands in vertebrate genomes . J. Mol. Biol. , 196 , 261 – 282 ( 1987. ). [DOI] [PubMed] [Google Scholar]

[b25] 25. ) Issa , J. P. , Ottaviano , Y. L. , Celano , P. , Hamilton , S. R. , Davidson , N. E. and Baylin , S. B.Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon . Nat. Genet , 7 , 536 – 540 ( 1994. ). [DOI] [PubMed] [Google Scholar]

[b26] 26. ) Miyakura , Y. , Sugano , K. , Konishi , F. , Ichikawa , A. , Maekawa , M. , Shitoh , K. , Igarashi , S. , Kotake , K. , Koyama , Y. and Nagai , H.Extensive methylation of hMLH1 promoter region predominates in proximal colon cancer with microsatellite instability . Gastroenterology , 121 , 1300 – 1309 ( 2001. ). [DOI] [PubMed] [Google Scholar]

[b27] 27. ) Ushijima , T. , Makino , H. , Nakayasu , M. , Aonuma , S. , Takeuchi , M. , Segawa , K. , Sugimura , T. and Nagao , M.Presence of p53 mutations in 3Y1‐B clone 1–6: a rat cell line widely used as a normal immortalized fibroblast . Jpn. J. Cancer Res. , 85 , 455 – 458 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b28] 28. ) Huschtscha , L. I. and Reddel , R. R.p16^INIK4a and the control of cellular proliferative life span . Cardnogenesis , 20 , 921 – 926 ( 1999. ). [DOI] [PubMed] [Google Scholar]

[b29] 29. ) Wong , D. J. , Foster , S. A. , Galloway , D. A. and Reid , B. J.Progressive region‐specific de novo methylation of the p16 CpG island in primary human mammary epithelial cell strains during escape from M(0) growth arrest . Mol. Cell. Biol. , 19 , 5642 – 5651 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

[b30] 30. ) Hui , R. , Macmillan , R. D. , Kenny , F. S. , Musgrove , E. A. , Blarney , R. W. , Nicholson , R. I. , Robertson , J. F. and Sutherland , R. L.INK4a gene expression and methylation in primary breast cancer: overexpression of p16^INK4a messenger RNA is a marker of poor prognosis . Clin. Cancer Res. , 6 , 2777 – 2787 ( 2000. ). [PubMed] [Google Scholar]

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Cloning of the 5’Upstream Region of the Rat p16 Gene and Its Role in Silencing

Masanobu Abe

Eriko Okochi

Takashi Kuramoto

Atsushi Kaneda

Tsuyoshi Takato

Takashi Sugimura

Toshikazu Ushijima

Abstract

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Cloning of the 5’Upstream Region of the Rat p16 Gene and Its Role in Silencing

Masanobu Abe

Eriko Okochi

Takashi Kuramoto

Atsushi Kaneda

Tsuyoshi Takato

Takashi Sugimura

Toshikazu Ushijima

Abstract

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