Expression of voltage-gated sodium channel in primary tumors and corresponding metastases in the Copenhagen rat model of prostate cancer

Abstract

Objective: Increasing evidence indicates that prostate cancer (PCa) cells possess voltage-gated sodium channels (VGSCs) and these channels contribute to the metastatic progression of the disease. In the present study, it was aimed to investigate the heterotopic expression of VGSCs in carcinogenesis and metastasis. It was evaluated whether the expression of VGSC in strongly metastatic rat Mat-LyLu PCa cells observed in vitro is also conserved when these cells form primary tumors and metastasize to the lungs upon their subcutaneous injection in rats. Methods: PCa model was induced in male Copenhagen rats by subcutaneous implantation of strongly metastatic Mat-LyLu cells. The expression of VGSCs was studied in the model utilizing molecular biological and immunochemical techniques. Results: Reverse transcription PCR and immunohistochemistry showed that the expression of VGSC is conserved in the primary prostate tumors of Mat-LyLu and not eliminated in the corresponding lung metastases at both the mRNA and protein level. Conclusion: The results are consistent with the predominant role of VGSC in the metastatic progression of PCa and consistently support the VGSC hypothesis of cancer progression. J Clin Exp Invest 2013; 4 (4): 422-428

Keywords

• Prostate cancer, voltage-gated sodium channel, primary tumor, metastasis

Prostat kanseri Copenhagen sıçan modelinin primer tümörleri ve metastazlarında voltaj kapılı sodyum kanal ekspresyonu

Öz

Amaç: Artan miktardaki kanıt prostat kanser (PCa) hücrelerinin voltaj kapılı sodyum kanalları (VGSCs)\'na sahip olduğunu ve bu kanalların hastalığın metastatik ilerlemesine katkıda bulunduğunu göstermektedir. Mevcut çalışmada, VGSC\'lerin karsigonezde ve metastazdaki heterotopik ekspresyonunu araştırmak amaçlanmıştır. Güçlü metastatik sıçan Mat-LyLu PCa hücrelerinde in vitro olarak gözlemlenen VGSC ekspresyonunun, bu hücrelerin sıçanlara subkuten enjekte edildiğinde primer tümör oluştuğunda ve hücreler akciğerlere metastaz yaptığında korunup korunmadığı, değerlendirilmiştir. Yöntemler: PCa modeli Copenhagen ırkı erkek sıçanlarda güçlü metastatik Mat-LyLu hücrelerinin subkuten implantasyonu ile oluşturulmuştur. VGSC eksprsyonu bu modelde moleküler biyolojik ve immünokimyasal yöntemlerle araştırılmıştır. Bulgular: Ters transkripsiyon PCR ve immünohistokimya yöntemleri VGSC ekspresyonunun Mat-LyLu primer prostat tümörlerinde korunduğunu ve akciğer metatsazlarında da bu ekspresyonun devam ettiğini, mRNA ve protein düzeyinde göstermiştir. Sonuç: Sonuçlar, VGSC\'lerin PCa\'nin metastatik ilerlemesindeki baskın rolü ile uyumludur ve ‘kanserin ilerlemesinde VGSC\'lerin rolü\' hipotezini tutarlı bir şekilde desteklemektedir.

Anahtar Kelimeler

• Prostat kanseri, voltaj-kapılı sodyum kanalı, primer tümör, metastaz

References

1. Fitzpatrick JM, Schulman C, Zlotta AR, Schröder FH. Prostate cancer: a serious disease suitable for pre- vention. BJU Int T 2009;103:864-870.
2. Grimes JA, Fraser SP, Stephens GJ, et al. Differen- tial expression of voltage-activied Na+ currents in two prostatic tomour cell lines: contribution to invasive- ness in vitro. FEBS Letters 1995;369:290-294.
3. Laniado ME, Lalani EN, Fraser SP, et al. Expression and functional analysis of voltage-activated Na+ channels in human prostate cancer cell lines and their contrubition to invasion in vitro. Am J Pathol 1997;150:1213-1221.
4. Diss JK, Archer SN, Hirano J, et al. Expression pro- files voltage gated Na+ channel alpha-subunit genes in rat and human prostate cancer cell lines. Prostate 2001;48:165-178.
5. Nakajima T, Kubota N, Tsutsumi T, et al. Eicosapentae- noic acid inhibits voltage-gated sodium channels and invasiveness in prostate cancer cells. Br J Pharmacol 2009;156:420-431.
6. Abdul M, Hoosein N. Voltage-gated sodium ion chan- nels in prostate cancer: expression and activity. Anti- cancer Research 2002;22:1727-1730.
7. Diss JK, Stewart D, Pani F, et al. A potential novel marker for human prostate cancer: Voltage-gated so- dium channel expression in vivo. Prostate Cancer & Prostatic Diseases 2005;8:266-273.
8. Roger S, Besson P, Guennec J. Involvement of a novel fast inward sodium current in the invasion capacity of a breast cancer cell line. Biochimica et Biophysica Acta 2003;1616:107-111.

9. Fraser SP, Diss JKJ, Chioni A-M, et al. Voltage-gated sodium channel expression and potentiation of hu- man breast cancer metastasis. Clinical Cancer Re- search 2005;11:5381-5389.
10. Onganer PU, Djamgoz MBA. Small-cell lung cancer (human): Potentiation of endocytic membrane activ- ity by voltage gated Na+ channel expression in vitro. Journal of Membrane Biology 2005;204:67-75.
11. Roger S, Rollin J, Barascu A, et al. Voltage-gated sodium channels potentiate the invasive capaci- ties of human non-small-cell lung cancer cell lines. International Journal of Biochemistry & Cell Biology 2007;39:774-786.
12. Allen DH, Lepple-Wienhues A, Cahalan MD. Ion chan- nel phenotype of melanoma cell lines. J Membrane Biol 1997;155:27-34.
13. Schrey M, Codina C, Kraft R, et al. Molecular charac- terization of voltage-gated sodium channels in human gliomas. Neuroreport 2002;20;13:2493-2498.
14. Fulgenzi G, Graciotti L, Faronato M, et al. Human neoplastic mesothelial cells express voltage-gated sodium channels involved in cell motility. The In- ternational Journal of Biochemistry & Cell Biology 2006;38:1146-1159.
15. Gao R, Shen Y, Cai J, et al. Expression of voltage-gat- ed sodium channel subunit in human ovarian cancer. Oncology Reports 2010;23:1293-1299.
16. House CD, Vaske CD, Schwartz AM, et al. Voltage- gated Na+ channel SCN5A is a key regulator of a gene transcriptional network that controls colon can- cer inavasion. Cancer Res 2010;70:6957-6967.
17. Hernandez-Plata E, Ortiz CS, Marquina-Castillo B, et al. Over expression of Nav1.6 channels is associated with the invasion capacity of human cervical cancer. Inter J Cancer 2011;Doi:10.1002/ijc.26210.
18. Bennett ES, Smith BA, Harper JM. Voltage-gated Na+ channels confer invasive properties on human prostate cancer cells. Pflugers Arch – Eur Physiol 2004;447:908-914.
19. Gillet L, Roger S, Besson P, et al. Voltage-gated so- dium channel activity promotes cysteine cathepsin- dependent invasiveness and colony growth of human cancer cells. J. Biol. Chem 2009;284:8680–8691.
20. Djamgoz MBA, Mycielska M, Madeja Z, et al. Direc- tional movement of rat prostate cancer cells in direct- current electric field: involvement of voltage-gated Na+ channel activity. J Cell Sci 2001;114:2697-2705.
21. Fraser SP, Salvador V, Manning E, et al. Contribution of functional voltage-gated Na+ channel expression to cell behaviours involved in the metastic cascade in rat prostate cancer: I. Lateral motility. J Cell Physiol 2003;169:479-487.
22. Fraser SP, Ding Y, Liu A, et al. Tetrodotoxin sup- presses morphological enhancement of the metastic Mat-LyLu rat prostate cancer cell line. Cell Tissue Res 1999;295:505-512.
23. Krasowska M, Grzywna ZJ, Mycielska ME, Djamgoz MBA. Patterning of endocytotic vesicles and its con- trol by voltage-gated Na+ channel activity in rat pros- tate cancer cells: fractal analyses. Eur Biophys J 2004;33:535–542.
24. Palmer CP, Mycielska ME, Burcu H, et al. Single cell adhesion measuring apparatus (SCAMA): application to cancer cell lines of different metastatic potential and voltage-gated Na+ channel expression. Eur Biophys- ics J 2008;37:359-368.
25. Brackenbury W, Djamgoz MBA. Activity-dependent regulation of voltage-gated Na+ channel expression in Mat-LyLu rat prostate cancer cell line. J Physiology 2006;573:343-356.
26. Brackenbury WJ, Chioni AM, Diss JKJ, Djamgoz MBA. The neonatal splice variant of Nav1.5 potenti- ates in vitro metastatic behaviour of MDA-MB-231 human breast cancer cells. Breast Cancer Res Treat 2007;101:149–160.
27. Khanna C, Hunter K. Modelling metastasis in vivo. Carcinogenesis 2005;26:513-523.
28. Yildirim S, Altun S, Gumushan H, et al. Voltage-gated sodium channel activity promotes prostate cancer metastasis in vivo. Cancer Letters 2012;323:58-61.
29. Hollingshead MG. Antitumor efficacy testing in ro- dents. J Natl Cancer Inst 2008;100:1500-1510.
30. Steeg PS, Theodorescu D. Metastasis: a therapeutic target for cancer. Nat Clin Pract Oncol 2008;5:206– 219.
31. Hunter KW, Crawford NP, Alsarraj JJ. Mechanisms of metastasis. Breast Cancer Res 2008;10:1:S2.
32. Nguyen DX, Massague J. Genetic determinants of cancer metastasis. Nature Reviews Genetics 2007;8:341-352.