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The relationship between connexins, gap junctions, tissue architecture and tumour invasion, as studied in a novel in vitro model of HPV-16-associated cervical cancer progression

Abstract

Disruption of gap junctional intercellular communication (GJIC) and/or connexins (gap junction proteins) is frequently reported in malignant cell lines and tumours. Certain human papillomaviruses (HPV) associated with the development of cancers, especially of the cervix, have previously been reported to downregulate GJIC in vitro. There is also evidence for reduced gap junctions in cervical dysplasia. However, many squamous hyperproliferative conditions, including HPV-induced warts, often show extensive upregulation of certain connexins. The association between HPV and GJIC, and the mechanism and consequence of deregulated GJIC in cervical tumour progression, remains unclear. Therefore, using a variety of nonmalignant and malignant cell lines and an organotypic raft-culture system, we investigated the relationship between HPV, gap junctions and tumour progression. Established cervical tumour cell lines carrying HPV were unable to communicate via gap junctions (when assayed by dye-transfer techniques). This correlated with lack of connexin protein expression, while transfection with connexins 26 or 43 led to functional gap junction membrane plaques. On the other hand, immortal but nonmalignant cell lines that contained episomal or integrated HPV-16, but required feeder-layer and growth-factor support, were consistently well coupled, and expressed multiple connexins at membrane junctions. In vitro selection of feeder-layer and growth-factor-independent variants eventually lead to loss of GJIC, which correlated with loss of membrane and increased cytoplasmic connexin 43 localization. However, this was preceded by loss of differentiation and stromal invasion, as assayed on the organotypic raft-culture model. Using this model, a comparison between noncoupled, well-coupled and connexin-transfected cell lines revealed no firm correlation between GJIC and dysplasia, but GJIC appeared to favour increased stratification. These findings demonstrate that loss of GJIC is frequent, but appears to occur more as a consequence of, rather than being the cause of, epithelial dysplasia, and may be influenced by, but is not directly attributable to, HPV.

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References

  • Akhurst RJ and Derynck R . (2001). Trends Cell Biol., 11, S44–S51.

  • Asamoto M, Takahashi S, Imaida K, Shirai T and Fukushima S . (1994). Carcinogenesis, 15, 2163–2166.

  • Ashrafi GH, Pitts JD, Faccini A, McLean P, O'Brien V, Finbow ME and Campo S . (2000). J. Gen. Virol., 81 (Part 3), 689–694.

  • Bhattacharyya T, Edward M, Cordery C and Richardson MD . (1998). Med. Mycol., 36, 15–19.

  • Brissette JL, Kumar NM, Gilula NB and Dotto GP . (1991). Mol. Cell. Biol., 11, 5364–5371.

  • Budunova IV, Carbajal S and Slaga TJ . (1995). Carcinogenesis, 16, 2717–2724.

  • Budunova IV, Carbajal S and Slaga TJ . (1996). Mol. Carcinogen., 15, 202–214.

  • DiMaio D and Mattoon D . (2001). Oncogene, 20, 7866–7873.

  • Dubina MV, Iatckii NA, Popov DE, Vasil'ev SV and Krutovskikh VA . (2002). Oncogene, 21, 4992–4996.

  • Duflot-Dancer A, Mesnil M and Yamasaki H . (1997). Oncogene, 15, 2151–2158.

  • Eiberger J, Degen J, Romualdi A, Deutsch U, Willecke K and Sohl G . (2001). Cell Adhes. Commun., 8, 163–165.

  • Ennaji MM, Schwartz JL, Mealing G, Belbaraka L, Parker C, Parentaux M, Jouishomme H, Arella M, Whitfield JF and Phipps J . (1995). Cell. Mol. Biol. (Noisy.-le-grand), 41, 481–498.

  • Faccini AM, Cairney M, Ashrafi GH, Finbow ME, Campo MS and Pitts JD . (1996). J. Virol., 70, 9041–9045.

  • Finbow ME and Pitts JD . (1993). J. Cell. Sci., 106 (Part 2), 463–471.

  • Fitzgerald DJ, Fusenig NE, Boukamp P, Piccoli C, Mesnil M and Yamasaki H . (1994). Carcinogenesis, 15, 1859–1865.

  • Friedl F, Kimura I, Osato T and Ito Y . (1970). Proc. Soc. Exp. Biol. Med., 135, 543–545.

  • Goldstein DJ, Finbow ME, Andresson T, McLean P, Smith K, Bubb V and Schlegel R . (1991). Nature, 352, 347–349.

  • Graeber SH and Hulser DF . (1998). Exp. Cell Res., 243, 142–149.

  • Habermann H, Ray V, Habermann W and Prins GS . (2002). J. Urol., 167, 655–660.

  • Hernandez-Blazquez FJ, Joazeiro PP, Omori Y and Yamasaki H . (2001). Exp. Cell Res., 270, 235–247.

  • Hirschi KK, Xu CE, Tsukamoto T and Sager R . (1996). Cell Growth Differ., 7, 861–870.

  • Holden PR, McGuire B, Stoler A, Balmain A and Pitts JD . (1997). Carcinogenesis, 18, 15–21.

  • Huang RP, Fan Y, Hossain MZ, Peng A, Zeng ZL and Boynton AL . (1998). Cancer Res., 58, 5089–5096.

  • Ito A, Katoh F, Kataoka TR, Okada M, Tsubota N, Asada H, Yoshikawa K, Maeda S, Kitamura Y, Yamasaki H and Nojima H . (2000). J. Clin. Invest., 105, 1189–1197.

  • Jamieson S, Going JJ, D'Arcy R and George WD . (1998). J. Pathol., 184, 37–43.

  • Jeon S, Allen-Hoffmann BL and Lambert PF . (1995). J. Virol., 69, 2989–2997.

  • Jeon S and Lambert PF . (1995). Proc. Natl. Acad. Sci. USA, 92, 1654–1658.

  • Kelsell DP, Dunlop J and Hodgins MB . (2001). Trends Cell Biol., 11, 2–6.

  • King TJ, Fukushima LH, Donlon TA, Hieber AD, Shimabukuro KA and Bertram JS . (2000a). Carcinogenesis, 21, 311–315.

  • King TJ, Fukushima LH, Hieber AD, Shimabukuro KA, Sakr WA and Bertram JS . (2000b). Carcinogenesis, 21, 1097–1109.

  • Krutovskikh VA, Troyanovsky SM, Piccoli C, Tsuda H, Asamoto M and Yamasaki H . (2000). Oncogene, 19, 505–513.

  • Kumar NM and Gilula NB . (1996). Cell, 84, 381–388.

  • Laird DW, Fistouris P, Batist G, Alpert L, Huynh HT, Carystinos GD and Alaoui-Jamali MA . (1999). Cancer Res., 59, 4104–4110.

  • Lampe PD and Lau AF . (2000). Arch. Biochem. Biophys., 384, 205–215.

  • Lesueur F, Mesnil M, Delouvee A, Girault JM, Yamasaki H, Thiery JP and Jouanneau J . (2002). Biochem. Biophys. Res. Commun., 294, 108–115.

  • Lin JH, Yang J, Liu S, Takano T, Wang X, Gao Q, Willecke K and Nedergaard M . (2003). J. Neurosci., 23, 430–441.

  • Lucke T, Choudhry R, Thom R, Selmer IS, Burden AD and Hodgins MB . (1999). J. Invest. Dermatol., 112, 354–361.

  • Manthey D, Banach K, Desplantez T, Lee CG, Kozak CA, Traub O, Weingart R and Willecke K . (2001). J. Membr. Biol., 181, 137–148.

  • Mantovani F and Banks L . (2001). Oncogene, 20, 7874–7887.

  • Martin PE, Blundell G, Ahmad S, Errington RJ and Evans WH . (2001). J. Cell Sci., 114, 3845–3855.

  • Martyn KD, Kurata WE, Warn-Cramer BJ, Burt JM, TenBroek E and Lau AF . (1997). Cell Growth Differ., 8, 1015–1027.

  • McNutt NS, Hershberg RA and Weinstein RS . (1971). J. Cell Biol., 51, 805–825.

  • McNutt NS and Weinstein RS . (1969). Science, 165, 597–599.

  • Mesnil M, Krutovskikh V, Piccoli C, Elfgang C, Traub O, Willecke K and Yamasaki H . (1995). Cancer Res., 55, 629–639.

  • Moorby C and Patel M . (2001). Exp. Cell Res., 271, 238–248.

  • Munger K, Basile JR, Duensing S, Eichten A, Gonzalez SL, Grace M and Zacny VL . (2001). Oncogene, 20, 7888–7898.

  • Naus CC, Bani-Yaghoub M, Rushlow W and Bechberger JF . (1999). Adv. Exp. Med. Biol., 468, 373–381.

  • Oelze I, Kartenbeck J, Crusius K and Alonso A . (1995). J. Virol., 69, 4489–4494.

  • Omori Y and Yamasaki H . (1998). Int. J. Cancer, 78, 446–453.

  • Omori Y, Zaidan Dagli ML, Yamakage K and Yamasaki H . (2001). Mutat. Res., 477, 191–196.

  • Pattillo RA, Hussa RO, Story MT, Ruckert AC, Shalaby MR and Mattingly RF . (1977). Science, 196, 1456–1458.

  • Piechocki MP, Burk RD and Ruch RJ . (1999). Carcinogenesis, 20, 401–406.

  • Plotkin LI, Manolagas SC and Bellido T . (2001). J. Biol. Chem., 277, 8648–8657.

  • Qin H, Shao Q, Curtis H, Galipeau J, Belliveau DJ, Wang T, Alaoui-Jamali MA and Laird DW . (2002). J. Biol. Chem., 277, 29132–29138.

  • Richard G, Rouan F, Willoughby CE, Brown N, Chung P, Ryynanen M, Jabs EW, Bale SJ, DiGiovanna JJ, Uitto J and Russell L . (2002). Am. J. Hum. Genet., 70, 1341–1348.

  • Risek B, Pozzi A and Gilula NB . (1998). J. Cell Sci., 111 (Part 10), 1395–1404.

  • Saito T, Nishimura M, Kudo R and Yamasaki H . (2001). Int. J. Cancer, 93, 317–323.

  • Saito T, Schlegel R, Andresson T, Yuge L, Yamamoto M and Yamasaki H . (1998). Oncogene, 17, 1673–1680.

  • Saitoh M, Oyamada M, Oyamada Y, Kaku T and Mori M . (1997). Carcinogenesis, 18, 1319–1328.

  • Saunders MM, Seraj MJ, Li Z, Zhou Z, Winter CR, Welch DR and Donahue HJ . (2001). Cancer Res., 61, 1765–1767.

  • Sawey MJ, Goldschmidt MH, Risek B, Gilula NB and Lo CW . (1996). Mol. Carcinogen., 17, 49–61.

  • Shore L, McLean P, Gilmour SK, Hodgins MB and Finbow ME . (2001). Biochem. J., 357, 489–495.

  • Simon AM and Goodenough DA . (1998). Trends Cell Biol., 8, 477–483.

  • Stanley MA, Browne HM, Appleby M and Minson AC . (1989). Int. J. Cancer, 43, 672–676.

  • Sterling J, Stanley M, Gatward G and Minson T . (1990). J. Virol., 64, 6305–6307.

  • Tan LW, Bianco T and Dobrovic A . (2002). Carcinogenesis, 23, 231–236.

  • Temme A, Buchmann A, Gabriel HD, Nelles E, Schwarz M and Willecke K . (1997). Curr. Biol., 7, 713–716.

  • Thomsen P, van Deurs B, Norrild B and Kayser L . (2000). Oncogene, 19, 6023–6032.

  • Tomakidi P, Cheng H, Kohl A, Komposch G and Alonso A . (2000). Cell Tissue Res., 301, 323–327.

  • Trosko JE and Ruch RJ . (1998). Front. Biosci., 3, D208–D236.

  • van Geel M, van Steensel MA, Kuster W, Hennies HC, Happle R, Steijlen PM and Konig A . (2002). Br. J. Dermatol., 146, 938–942.

  • Vessey CJ, Wilding J, Folarin N, Hirano S, Takeichi M, Soutter P, Stamp GW and Pignatelli M . (1995). J. Pathol., 176, 151–159.

  • Wiszniewski L, Salomon D and Meda P . (2001). Cell Adhes. Commun., 8, 409–413.

  • Yamakage K, Omori Y, Piccoli C and Yamasaki H . (1998). Mol. Carcinogen., 23, 121–128.

  • Yamakage K, Omori Y, Zaidan-Dagli ML, Cros MP and Yamasaki H . (2000). J. Invest. Dermatol., 114, 289–294.

  • Yamasaki H, Krutovskikh V, Mesnil M, Tanaka T, Zaidan-Dagli ML and Omori Y . (1999). C. R. Acad. Sci. III, 322, 151–159.

  • Yano T, Hernandez-Blazquez FJ, Omori Y and Yamasaki H . (2001). Carcinogenesis, 22, 1593–1600.

  • Yeager M, Unger VM and Falk MM . (1998). Curr. Opin. Struct. Biol., 8, 517–524.

  • Zhai Y, Wu R, Schwartz DR, Darrah D, Reed H, Kolligs FT, Nieman MT, Fearon ER and Cho KR . (2002). Am. J. Pathol., 160, 1229–1238.

  • Zhang ZQ, Zhang W, Wang NQ, Bani-Yaghoub M, Lin ZX and Naus CC . (1998). Carcinogenesis, 19, 1889–1894.

  • zur Hausen H . (2002). Nat. Rev. Cancer, 2, 342–350.

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Acknowledgements

We are grateful to Dr Edgar Rivedal for the Cx43 antibody, Dr Paul Lambert for the W12E and W12G cell lines, Dr Margaret Stanley for the original W12 cell line, Dr Scott Cuthill for the SiHa and CaSki cell lines, Dr Ian Morgan for the C33a cell line, Dr Susan Jamieson and Dr Hiroshi Yamasaki for the HeLa and HeLa43 cells, and Dr Dale Laird for the Cx43-GFP plasmid. T Aasen was supported by a University of Glasgow PhD grant, and by Roche Products, UK.

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Correspondence to Malcolm B Hodgins.

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Aasen, T., Hodgins, M., Edward, M. et al. The relationship between connexins, gap junctions, tissue architecture and tumour invasion, as studied in a novel in vitro model of HPV-16-associated cervical cancer progression. Oncogene 22, 7969–7980 (2003). https://doi.org/10.1038/sj.onc.1206709

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