Connexin subtype expression during oral carcinogenesis: A pilot study in patients with oral squamous cell carcinoma

Brockmeyer,Phillipp; Hemmerlein,Bernhard; Jung,Klaus; Fialka,Florian; Brodmann,Tobias; Gruber,Rudolf Matthias; Schliephake,Henning; Kramer,Franz‑Josef

doi:10.3892/mco.2015.685

Connexin subtype expression during oral carcinogenesis: A pilot study in patients with oral squamous cell carcinoma

Authors:
- Phillipp Brockmeyer
- Bernhard Hemmerlein
- Klaus Jung
- Florian Fialka
- Tobias Brodmann
- Rudolf Matthias Gruber
- Henning Schliephake
- Franz‑Josef Kramer
View Affiliations

Affiliations: Department of Oral and Maxillofacial Surgery, University Medical Centre Goettingen, D‑37075 Goettingen, Germany, Department of Pathology, University Medical Centre Goettingen, D‑37075 Goettingen, Germany, Department of Medical Statistics, University Medical Centre Goettingen, D‑37075 Goettingen, Germany, Private Practice, MKG Leinetal, D‑37574 Einbeck, Germany, Private Practice, Zahnarztpraxis am Westertor, D‑37115 Duderstadt, Germany
Published online on: November 25, 2015 https://doi.org/10.3892/mco.2015.685
Pages: 298-302

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Gap junctional intercellular communication (GJIC) and connexin (Cx) expression were reported in association with carcinogenesis in various types of tumours. In an earlier histomorphometric study, the protein levels of Cx subtypes 26, 43 and 45 were differentially expressed in oral squamous cell carcinoma (OSCC), corresponding lymph node metastases and dysplasia‑free oral mucosa. Moreover, membrane Cx43 acted as an independent prognostic marker in OSCC tissues. This study aimed to confirm the expression of described Cx subtypes at the mRNA level. Hence, a reverse transcription quantitative polymerase chain reaction (RT‑qPCR) analysis of Cx26, Cx43 and Cx45 gene expressions was performed in paired carcinoma and mucosa samples of 15 OSCC patients. Additionally, we assessed the interaction between Cx subtype expression and clinicopathological routine parameters. The RT‑qPCR analysis revealed that Cx26 was downregulated in OSCC (P=0.01), while Cx43 was marginally upregulated in cancer tissue (P=0.04). Cx45 was significantly overexpressed in OSCC tissue compared with the intraoral mucosa controls (P<0.01), and remained unchanged at different tumour stages. No significant interactions between differential Cx subtype expression and clinicopathological routine parameters were observed. In conclusion, Cx regulation at the transcriptional level appears to be an early event during the initiation and development of OSCC, and is maintained during further progression. However, the mRNA‑protein correlation is variable. This may be indicative of post‑transcriptional, translational and degradation regulations being associated with the determination of Cx protein concentration during oral carcinogenesis.

View Figures

View References

1	Willecke K, Eiberger J, Degen J, Eckardt D, Romualdi A, Güldenagel M, Deutsch U and Söhl G: Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem. 383:725–737. 2002. View Article : Google Scholar : PubMed/NCBI
2	Loewenstein WR and Penn RD: Intercellular communication and tissue growth. II. Tissue regeneration. J Cell Biol. 33:235–242. 1967. View Article : Google Scholar : PubMed/NCBI
3	Mesnil M, Krutovskikh V, Piccoli C, Elfgang C, Traub O, Willecke K and Yamasaki H: Negative growth control of HeLa cells by connexin genes: Connexin species specificity. Cancer Res. 55:629–639. 1995.PubMed/NCBI
4	King TJ, Fukushima LH, Donlon TA, Hieber AD, Shimabukuro KA and Bertram JS: Correlation between growth control, neoplastic potential and endogenous connexin 43 expression in HeLa cell lines: Implications for tumor progression. Carcinogenesis. 21:311–315. 2000. View Article : Google Scholar : PubMed/NCBI
5	McLachlan E, Shao Q, Wang HL, Langlois S and Laird DW: Connexins act as tumor suppressors in three-dimensional mammary cell organoids by regulating differentiation and angiogenesis. Cancer Res. 66:9886–9894. 2006. View Article : Google Scholar : PubMed/NCBI
6	Avanzo JL, Mesnil M, Hernandez-Blazquez FJ, Mackowiak II, Mori CM, da Silva TC, Oloris SC, Gárate AP, Massironi SM, Yamasaki H and Dagli ML: Increased susceptibility to urethane-induced lung tumors in mice with decreased expression of connexin 43. Carcinogenesis. 25:1973–1982. 2004. View Article : Google Scholar : PubMed/NCBI
7	Dubina MV, Iatckii NA, Popov DE, Vasil'ev SV and Krutovskikh VA: Connexin 43, but not connexin 32, is mutated at advanced stages of human sporadic colon cancer. Oncogene. 21:4992–4996. 2002. View Article : Google Scholar : PubMed/NCBI
8	Cronier L, Crespin S, Strale PO, Defamie N and Mesnil M: Gap junctions and cancer: New functions for an old story. Antioxid Redox Signal. 11:323–338. 2009. View Article : Google Scholar : PubMed/NCBI
9	Trosko JE, Chang CC, Upham BL and Tai MH: Ignored hallmarks of carcinogenesis: Stem cells and cell-cell communication. Ann N Y Acad Sci. 1028:192–201. 2004. View Article : Google Scholar : PubMed/NCBI
10	Iacobas DA, Urban-Maldonado M, Iacobas S, Scemes E and Spray DC: Array analysis of gene expression in connexin-43 null astrocytes. Physiol Genomics. 15:177–190. 2003. View Article : Google Scholar : PubMed/NCBI
11	Iacobas DA, Scemes E and Spray DC: Gene expression alterations in connexin null mice extend beyond the gap junction. Neurochem Int. 45:243–250. 2004. View Article : Google Scholar : PubMed/NCBI
12	Yano T, Hernandez-Blazquez FJ, Omori Y and Yamasaki H: Reduction of malignant phenotype of HEPG2 cell is associated with the expression of connexin 26 but not connexin 32. Carcinogenesis. 22:1593–1600. 2001. View Article : Google Scholar : PubMed/NCBI
13	Betsuyaku T, Nnebe NS, Sundset R, Patibandla S, Krueger CM and Yamada KA: Overexpression of cardiac connexin 45 increases susceptibility to ventricular tachyarrhythmias in vivo. Am J Physiol Heart Circ Physiol. 290:H163–H171. 2006. View Article : Google Scholar : PubMed/NCBI
14	Bukauskas FF, Angele AB, Verselis VK and Bennett MV: Coupling asymmetry of heterotypic connexin 45/connexin 43-EGFP gap junctions: Properties of fast and slow gating mechanisms. Proc Natl Acad Sci USA. 99:7113–7118. 2002. View Article : Google Scholar : PubMed/NCBI
15	Koval M, Geist ST, Westphale EM, Kemendy AE, Civitelli R, Beyer EC and Steinberg TH: Transfected connexin 45 alters gap junction permeability in cells expressing endogenous connexin 43. J Cell Biol. 130:987–995. 1995. View Article : Google Scholar : PubMed/NCBI
16	Ogawa K, Pitchakarn P, Suzuki S, Chewonarin T, Tang M, Takahashi S, Naiki-Ito A, Sato S, Takahashi S, Asamoto M and Shirai T: Silencing of connexin 43 suppresses invasion, migration and lung metastasis of rat hepatocellular carcinoma cells. Cancer Sci. 103:860–867. 2012. View Article : Google Scholar : PubMed/NCBI
17	Saito-Katsuragi M, Asada H, Niizeki H, Katoh F, Masuzawa M, Tsutsumi M, Kuniyasu H, Ito A, Nojima H and Miyagawa S: Role for connexin 26 in metastasis of human malignant melanoma: Communication between melanoma and endothelial cells via connexin 26. Cancer. 110:1162–1172. 2007. View Article : Google Scholar : PubMed/NCBI
18	Ito A, Koma Y, Uchino K, Okada T, Ohbayashi C, Tsubota N and Okada M: Increased expression of connexin 26 in the invasive component of lung squamous cell carcinoma: Significant correlation with poor prognosis. Cancer Lett. 234:239–248. 2006. View Article : Google Scholar : PubMed/NCBI
19	Kanczuga-Koda L, Sulkowski S, Lenczewski A, Koda M, Wincewicz A, Baltaziak M and Sulkowska M: Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer. J Clin Pathol. 59:429–433. 2006. View Article : Google Scholar : PubMed/NCBI
20	Lin JH, Takano T, Cotrina ML, Arcuino G, Kang J, Liu S, Gao Q, Jiang L, Li F, Lichtenberg-Frate H, et al: Connexin 43 enhances the adhesivity and mediates the invasion of malignant glioma cells. J Neurosci. 22:4302–4311. 2002.PubMed/NCBI
21	Ozawa H, Matsunaga T, Kamiya K, Tokumaru Y, Fujii M, Tomita T and Ogawa K: Decreased expression of connexin-30 and aberrant expression of connexin-26 in human head and neck cancer. Anticancer Res. 27:2189–2195. 2007.PubMed/NCBI
22	Villaret DB, Wang T, Dillon D, Xu J, Sivam D, Cheever MA and Reed SG: Identification of genes overexpressed in head and neck squamous cell carcinoma using a combination of complementary DNA subtraction and microarray analysis. Laryngoscope. 110:374–381. 2000. View Article : Google Scholar : PubMed/NCBI
23	Brockmeyer P, Jung K, Perske C, Schliephake H and Hemmerlein B: Membrane connexin 43 acts as an independent prognostic marker in oral squamous cell carcinoma. Int J Oncol. 45:273–281. 2014.PubMed/NCBI
24	Xia J, Liu X, Tao X, Hong Y, Chen X, Dai Y, Huang Y and Cheng B: Expression of gap junctional protein connexin 43 during 4-nitroquinoline-1-oxide-induced rat tongue carcinogenesis. J Mol Histol. 40:183–188. 2009. View Article : Google Scholar : PubMed/NCBI
25	Wolff KD, Follmann M and Nast A: The diagnosis and treatment of oral cavity cancer. Dtsch Arztebl Int. 109:829–835. 2012.PubMed/NCBI
26	Fialka F, Gruber RM, Hitt R, Opitz L, Brunner E, Schliephake H and Kramer FJ: CPA6, FMO2, LGI1, SIAT1 and TNC are differentially expressed in early- and late-stage oral squamous cell carcinoma - a pilot study. Oral Oncol. 44:941–948. 2008. View Article : Google Scholar : PubMed/NCBI
27	Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e452001. View Article : Google Scholar : PubMed/NCBI
28	Holm S: A simple sequentially rejective multiple test procedure. Scandinavian J Stat. 6:65–70. 1979.
29	Zhang ZQ, Hu Y, Wang BJ, Lin ZX, Naus CC and Nicholson BJ: Effective asymmetry in gap junctional intercellular communication between populations of human normal lung fibroblasts and lung carcinoma cells. Carcinogenesis. 25:473–482. 2004. View Article : Google Scholar : PubMed/NCBI
30	Udaka N, Miyagi Y and Ito T: Connexin expression in mouse lung tumor. Cancer Lett. 246:224–229. 2007. View Article : Google Scholar : PubMed/NCBI
31	Yamada KA, Rogers JG, Sundset R, Steinberg TH and Saffitz J: Up-regulation of connexin 45 in heart failure. J Cardiovasc Electrophysiol. 14:1205–1212. 2003. View Article : Google Scholar : PubMed/NCBI
32	Xing Y, Xiao Y, Zeng F, Zhao J, Xiao C, Xiong P and Feng W: Altered expression of connexin-43 and impaired capacity of gap junctional intercellular communication in prostate cancer cells. J Huazhong Univ Sci Technolog Med Sci. 27:291–294. 2007. View Article : Google Scholar : PubMed/NCBI
33	Vogel C and Marcotte EM: Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet. 13:227–232. 2012.PubMed/NCBI
34	Budunova IV, Carbajal S and Slaga TJ: The expression of gap junctional proteins during different stages of mouse skin carcinogenesis. Carcinogenesis. 16:2717–2724. 1995. View Article : Google Scholar : PubMed/NCBI