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Inhibiting tumorigenic potential by restoration of p16 in nasopharyngeal carcinoma

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Summary

The p16 gene, encodes a key checkpoint protein p16 in the cell cycle, has been reported inactivation in a wide variety of human cancers. We have previously demonstrated high frequency of p16 alterations in primary nasopharyngeal carcinoma (NPC), xenografts and cell lines. The finding implied that inactivation of the p16 gene may play an important role in the NPC development. To investigate the tumour suppressor function of p16 in NPC, we tranfected p16-deficient NPC cell line, NPC/HK-1, with a wild-type p16 expression construct, and evaluated growth and tumorigenic properties of the clones stably expressing exogenous p16. Expression of the exogenous wild-type p16 significantly inhibited cell growth by more than 70% when compared to that of the parental and empty vector-transfected cells. This growth inhibition was attributable to a significant proportion of p16-expressing cells arrested at G1 phase in the cell cycle as revealed by flow cytometric analysis. By anchorage-independent colony forming assay, we found that the ability to form colonies in soft agar was highly reduced in cells expressing p16. NPC/HK1 cells expressing functional p16 also showed suppressed tumorigenicity in athymic nude mice. Taken together, our results provide strong evidence for a tumour suppressor role of p16 in NPC.

Change history

  • 16 November 2011

    This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication

References

  • Castellano, M, Gabrielli, BG, Hussussian, CJ, Dracopoli, NC & Hayward, NK (1997). Restoration of CDKN2A into melanoma cells induces morphologic changes and reduction in growth rate but not anchorage-idependent growth reversal. J Invest Dermatol 109: 61–68.

    CAS  Article  Google Scholar 

  • Chan, FKM, Zhang, J, Chen, L, Shapiro, DN & Winoto, A (1995). Indentification of human/mouse p19, a novel cdk4/cdk6 inhibitor with homology to p16ink4. Mol Cell Biol 15: 2682–2688.

    CAS  Article  Google Scholar 

  • Cheng, RYS, Lo, KW, Huang, DP & Tsao, SW (1997). Loss of heterozygosity on chromosome 14 in primary nasopharyngeal carcinoma. Int J Oncol 10: 1047–1050.

    CAS  PubMed  Google Scholar 

  • Cheng, Y, Poulos, NE, Lung, ML, Hampton, G, Ou, B, Lerman, MI & Stanbridge, EJ (1998). Functional evidence for a nasopharyngeal carcinoma tumor suppressor gene that maps at chromosome 3p21.3. Proc Natl Acad Sci USA 95: 3042–3047.

    CAS  Article  Google Scholar 

  • Fueyo, J, Gomez-Manzano, C, Yung, WKA, Clayman, GL, Liu, TJ, Bruner, J, Levin, VA & Kyritsis, AP (1996). Adenovirus-mediated p16/CDKN2 gene transfer induces growth arrest and modifies the transformed phenotype of glioma cells. Oncogene 12: 103–110.

    CAS  PubMed  Google Scholar 

  • Gulley, ML, Nicholls, JM, Schneider, BG, Amin, MB, Ro, JY & Geradts, J (1998). Nasopharyngeal carcinomas frequently lack the p16/MTS1 tumor suppressor protein but consistently express the retinoblastoma gene product. Am J Pathol 152: 865–869.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hannon, GJ & Beach, D (1994). p15INK4b is a potential effector of cell cycle arrest mediated by TGF-β. Nature (Lond) 371: 257–261.

    CAS  Article  Google Scholar 

  • Hinds, PS, Mittnacht, S, Dulic, V, Arnold, A, Reed, SI & Weinberg, RA (1992). Regulation of retinoblastoma protein functions by ectopic expression of human cyclins. Cell 70: 993–1006.

    CAS  Article  Google Scholar 

  • Huang, DP, Ho, JHC, Poon, YF, Chew, EC, Saw, D, Liu, M, Li, CL, Mak, LS, Lai, SH & Lau, EH (1980). Establishment of a cell line (NPC/HK1) from a differentiated squamous carcinomas of the nasopharynx. Int J Cancer 26: 127–132.

    CAS  Article  Google Scholar 

  • Huang, DP, Lo, KW, Choi, PHK, Ng, AYT, Tsao, SY, Yiu, GKC & Lee, JC (1991). Loss of heterozygosity on the short arm of chromosome 3 in nasopharyngeal carcinoma. Cancer Genet Cytogenet 54: 91–99.

    CAS  Article  Google Scholar 

  • Huang, DP, Lo, KW, van Hasselt, CA, Woo, JK, Choi, PH, Leung, SF, Cheung, ST, Cairns, O, Sidransky, D & Lee, JC (1994). A region of homozygous deletion on chromosome 9p21–22 in primary nasopharyngeal carcinoma. Cancer Res 54: 4003–4006.

    CAS  PubMed  Google Scholar 

  • Hui, ABY, Lo, KW, Leung, SF, Choi, PH, Fong, Y, Lee, JC & Huang, DP (1996). Loss of heterozygosity on the long arm of chromosome 11 in nasopharyngeal carcinoma. Cancer Res 56: 3225–3229.

    CAS  PubMed  Google Scholar 

  • Jin, X, Nguyen, D, Zhang, WW, Kyritsis, AP & Roth, JA (1995). Cell cycle arrest and inhibition of tumor cell proliferation by the p16INK4 gene mediated by an adenovirus vector. Cancer Res 55: 3250–3253.

    CAS  PubMed  Google Scholar 

  • Kamb, A, Gruis, A, Weaver-Feldhaus, J, Liu, QY, Harshman, K, Tavtigian, SV, Stockert, E, Day, III RS, Johnson, BE & Skolnick, MH (1994). A cell cycle regulator potentially involved in genesis of many tumor types. Science 264: 436–439.

    CAS  Article  Google Scholar 

  • Kato, JY, Matsushime, H, Hiebert, SW, Ewen, ME & Sherr, CJ (1993). Direct binding of cyclin D to the retinoblastoma gene product (pRb) and pRb phosphorylation by the cyclin D-dependent kinase CDK4. Gene Dev 7: 331–342.

    CAS  Article  Google Scholar 

  • Li, JH, Li, P, Klamut, H & Liu, FF (1997). Cytotoxic effects of ad5CMV-p53 expression in two human nasopharyngeal carcinoma cell lines. Clin Cancer Res 3: 507–514.

    CAS  PubMed  Google Scholar 

  • Liggett, WH Jr, Sewell, DA, Rocco, J, Ahrendt, SA, Koch, W & Sidransky, D (1996). p16 and p16 are potent growth suppressors of head and neck squamous carcinoma cells in vitro. Cancer Res 56: 4119–4123.

    CAS  PubMed  Google Scholar 

  • Lo, KW, Huang, DP & Lau, KM (1995). p16 gene alterations in nasopharyngeal carcinoma. Cancer Res 55: 2039–2043.

    CAS  PubMed  Google Scholar 

  • Lo, KW, Cheung, ST, Leung, SF, van Hasselt, A, Tsang, YS, Mak, KF, Chung, YF, Woo, JKS, Lee, JCK & Huang, DP (1996). Hypermethylation of the p16 gene in nasopharyngeal carcinoma. Cancer Res 56: 2721–2725.

    CAS  PubMed  Google Scholar 

  • Lo, KW, Huang, DP & Lee, JCK (1997). Genetic changes in nasopharyneal carcinoma (NPC). Chinese Med J (Beijing) 110: 548–559.

    CAS  Google Scholar 

  • Lu, QL, Elia, G, Lucas, S & Thoma, JA (1993). Bcl-2 proto-oncogene expression in Epstein-Barr-virus-associated nasopharyngeal carcinoma. Int J Cancer 53: 29–35.

    CAS  Article  Google Scholar 

  • Miyakoshi, J, Kitagawa, K, Yamagishi, N, Ohtsu, S, Day, RSIII & Takebe, H (1997). Increased radiosensitivity of p16 gene-deleted human glioma cells after transfection with wild-type p16 gene. Jpn J Cancer Res 88: 34–38.

    CAS  Article  Google Scholar 

  • Nobori, T, Miura, K, Wu, DJ, Lois, A, Takabaryashi, K & Carson, DA (1994). Deletions of the cyclin-dependent kinase-4 inhibitor gene in multiple human cancers. Nature (Lond) 368: 753–756.

    CAS  Article  Google Scholar 

  • Pagano, M, Pepper, R, Lukas, J, Baldin, V, Ansorge, W, Bartek, J & Draetta, G (1993). Regulation of the cell cycle by the cdk2 protein kinase in cultured human fibroblasts. J Cell Biol 121: 110–111.

    Article  Google Scholar 

  • Porter, MJ, Field, JK, Leung, SF, Lo, D, Lee, JC, Spandidos, DA & van Hasselt, CA (1994). The detection of the c-myc and ras oncogenes in nasopharyngeal carcinoma by immunohistochemistry. Acta Otolaryngol 114: 105–109.

    CAS  Article  Google Scholar 

  • Quesnel, B, Preudhomme, C, Lepelley, P, Hetuin, D, Vanrumbeke, M, Bauters, F, Velu, T & Fenaux, P (1996). Transfer of p16inka/CDKN2 gene in leukaemic cell lines inhibits cell proliferation. Br J Haematol 95: 291–298.

    CAS  Article  Google Scholar 

  • Spillare, E, Okamoto, A, Hagiwara, K, Demetrick, DJ, Serrano, M, Beach, D & Harris, CC (1996). Suppression of growth in vitro and tumorigenicity in vivo of human carcinoma cell lines by transfected p16INK4. Mol Carcinogen 16: 53–60.

    CAS  Article  Google Scholar 

  • Spruck, III CH, Tsai, YC, Huang, DP, Yang, AS, Rideout, III WM, Gonzalez-Zulueta, M, Choi, P, Lo, KW, Yu, MC & Jones, PA (1992). Absence of p53 gene mutations in primary nasopharyngeal carcinomas. Cancer Res 52: 4787–4790.

    CAS  PubMed  Google Scholar 

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Wang, G., Lo, K., Tsang, K. et al. Inhibiting tumorigenic potential by restoration of p16 in nasopharyngeal carcinoma. Br J Cancer 81, 1122–1126 (1999). https://doi.org/10.1038/sj.bjc.6690818

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Keywords

  • nasopharyngeal carcinoma
  • p16
  • tumour suppressor
  • gene transfer

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