The deregulation of claudin-3 has been reported to correlate with the invasion and metastasis of various cancers, but little is known about its expression level and the prognostic value in squamous cell lung carcinoma (SqCC). The purpose of this study is to determine the expression levels and the prognostic value of claudin-3 in completely resected SqCC tissues, and the potential underlying mechanism. The protein expression of claudin-3, E-cadherin, β-catenin, and vimentin in the tumor tissues from 103 patients with surgically resected SqCC was examined using immunohistochemistry, western blots, as well as semi-quantitative estimation. The claudin-3 protein level was significantly associated with E-cadherin, β-catenin, and vimentin protein expression. A decreased claudin-3 protein level was significantly correlated with TNM stage, lymph node metastasis, and disease recurrence. Similarly, downregulation of E-cadherin was significantly correlated with lymph node metastasis and disease recurrence. Decreased β-catenin expression also had a significant correlation with disease recurrence. Univariate analyses indicated that the T stage, lymph node metastasis, the TNM stage, and the expression of claudin-3, β-catenin, and vimentin were significant predictors for overall survival (OS). Moreover, multivariate analyses demonstrated that the TNM stage and protein levels of claudin-3, β-catenin, and vimentin were independent predictors for OS of SqCC patients. Claudin-3 plays an important role in the epithelial–mesenchymal transition of SqCC and might be used as a potential prognostic factor for SqCC.
This is a preview of subscription content, log in to check access.
This work was supported by Grants No. 81272615 from the Natural Science Foundation of China (to Bangwei Cao), No. 2011-3-007 from the Beijing Municipal Health System High-level Health Person Foundation Project (to Bangwei Cao), and No. yyqdkt2014-11 from the Beijing Friendship Hospital Scientific Research Foundation Project (to Juanjuan Che).
Juanjuan Che performed the experiments and wrote the manuscript. Yifan Yang, Jing Xiao, and Pengfei Zhao designed the experiment and collected the tumor samples. Bo Yan and Shuo Dong analyzed the experimental data. Bangwei Cao directed the work and edited the manuscript.
Conflicts of interest
The study was approved by the Research Ethics Committee of Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Data sharing statement
The authors declare that they have no conflicts of interest. This work is original and not under consideration for publication elsewhere.
Soini Y. Tight junctions in lung cancer and lung metastasis. A review. Int J Clin Exp Pathol. 2012;5:126–36. PubMed: 22400072.PubMedGoogle Scholar
Moore W, Talati R, Bhattacharji P, Bilfinger T. Five-year survival after cryoablation of stage I non-small cell lung cancer in medically inoperable patients. J Vasc Interv Radiol. 2015;26:312–9. PubMed: 25735518.CrossRefPubMedGoogle Scholar
Dempke WC, Suto T, Reck M. Targeted therapies for non-small cell lung cancer. Lung Cancer. 2010;67:257–74. PubMed: 19914732.CrossRefPubMedGoogle Scholar
Tokumo M, Toyooka S, Kiura K, Shigematsu H, Tomii K, Aoe M, et al. The relationship between epidermal growth factor receptor mutations and clinicopathologic features in non–small cell lung cancers. Clin Cancer Res. 2005;11:1167–73. PubMed: 15709185.PubMedGoogle Scholar
Tsao MS, Sakurada A, Cutz JC, Zhu CQ, Kamel-Reid S, Squire J, et al. Erlotinib in lung cancer—molecular and clinical predictors of outcome. N Engl J Med. 2005;353:133–44. PubMed: 16014883.CrossRefPubMedGoogle Scholar
Nichols LS, Ashfaq R, Iacobuzio-Donahue CA. Claudin 4 protein expression in primary and metastatic pancreatic cancer support for use as a therapeutic target. Am J Clin Pathol. 2004;121:226–30. PubMed: 14983936.CrossRefPubMedGoogle Scholar
Resnick MB, Konkin T, Routhier J, Sabo E, Pricolo VE. Claudin-1 is a strong prognostic indicator in stage II colonic cancer: a tissue microarray study. Mod Pathol. 2005;18:511–8. PubMed: 15475928.CrossRefPubMedGoogle Scholar
Szasz AM, Nemeth Z, Gyorffy B, Micsinai M, Krenacs T, Baranyai Z, et al. Identification of a claudin-4 and E-cadherin score to predict prognosis in breast cancer. Cancer Sci. 2011;102:2248–54. PubMed: 21883696.CrossRefPubMedGoogle Scholar
Shang X, Lin X, Alvarez E, Manorek G, Howell SB. Tight junction proteins claudin-3 and claudin-4 control tumor growth and metastases. Neoplasia. 2012;14:974–85. PubMed: 23097631.CrossRefPubMedPubMedCentralGoogle Scholar
Chao YC, Pan SH, Yang SC, Yu SL, Che TF, Lin CW, et al. Claudin-1 is a metastasis suppressor and correlates with clinical outcome in lung adenocarcinoma. Am J Respir Crit Care Med. 2009;179:123–33. PubMed: 18787218.CrossRefPubMedGoogle Scholar
Heinzelmann-Schwarz VA, Gardiner-Garden M, Henshall SM, Scurry J, Scolyer RA, Davies MJ, et al. Overexpression of the cell adhesion molecules DDR1, claudin 3, and Ep-CAM in metaplastic ovarian epithelium and ovarian cancer. Clin Cancer Res. 2004;10:4427–36. PubMed: 15240533.CrossRefPubMedGoogle Scholar
Merikallio H, Kaarteenaho R, Pääkkö P, Lehtonen S, Hirvikoski P, Mäkitaro R, et al. Impact of smoking on the expression of claudins in lung carcinoma. Eur J Cancer. 2011;47:620–30. PubMed: 21106366.CrossRefPubMedGoogle Scholar
Moldvay J, Jäckel M, Páska C, Soltész I, Schaff Z, Kiss A. Distinct claudin expression profile in histologic subtypes of lung cancer. Lung Cancer. 2007;57:159–67. PubMed: 17418912.CrossRefPubMedGoogle Scholar
Gao F, Duan X, Lu X, Liu Y, Zheng L, Ding Z, et al. Novel binding between pre-membrane protein and claudin-1 is required for efficient dengue virus entry. Biochem Biophys Res Commun. 2010;391:952–7. PubMed: 19962368.CrossRefPubMedGoogle Scholar
Van Itallie CM, Betts L, Smedley JG, McClane BA, Anderson JM. Structure of the claudin-binding domain of Clostridium perfringens enterotoxin. J Biol Chem. 2008;283:268–74. PubMed: 17977833.CrossRefPubMedGoogle Scholar
Saeki R, Kondoh M, Kakutani H, Matsuhisa K, Takahashi A, Suzuki H, et al. A claudin-targeting molecule as an inhibitor of tumor metastasis. J Pharmacol Exp Ther. 2010;334:576–82. PubMed: 20442222.CrossRefPubMedGoogle Scholar
Kupferman ME, Jiffar T, El-Naggar A, Yilmaz T, Zhou G, Xie T, et al. TrkB induces EMT and has a key role in invasion of head and neck squamous cell carcinoma. Oncogene. 2010;29:2047–59. PubMed: 20101235.CrossRefPubMedPubMedCentralGoogle Scholar
Iwatsuki M, Mimori K, Yokobori T, Ishi H, Beppu T, Nakamori S, et al. Epithelial–mesenchymal transition in cancer development and its clinical significance. Cancer Sci. 2010;101:293–9. PubMed: 19961486.CrossRefPubMedGoogle Scholar
Kase S, Sugio K, Yamazaki K, Okamoto T, Yano T, Sugimachi K. Expression of E-cadherin and beta-catenin in human non-small cell lung cancer and the clinical significance. Clin Cancer Res. 2000;6:4789–96. PubMed: 11156236.PubMedGoogle Scholar
Choi YS, Shim YM, Kim S-H, Son DS, Lee H-S, Kim GY, et al. Prognostic significance of E-cadherin and beta-catenin in resected stage I non-small cell lung cancer. Eur J Cardiothorac Surg. 2003;24:441–9. PubMed: 12965318.CrossRefPubMedGoogle Scholar
Chiu CG, Chan SK, Fang ZA, Masoudi H, Wood-Baker R, Jones SJ, et al. Beta-catenin expression is prognostic of improved non-small cell lung cancer survival. Am J Surg. 2012;203:654–9. PubMed: 22402266.CrossRefPubMedGoogle Scholar
Richardson F, Young GD, Sennello R, Wolf J, Argast GM, Mercado P, et al. The evaluation of E-cadherin and vimentin as biomarkers of clinical outcomes among patients with non-small cell lung cancer treated with erlotinib as second- or third-line therapy. Anticancer Res. 2012;32:537–52. PubMed: 22287743.PubMedGoogle Scholar
Schmalhofer O, Brabletz S, Brabletz T. E-cadherin, beta-catenin, and ZEB1 in malignant progression of cancer. Cancer Metastasis Rev. 2009;28:151–66. PubMed: 19153669.CrossRefPubMedGoogle Scholar
De Wever O, Pauwels P, De Craene B, Sabbah M, Emami S, Redeuilh G, et al. Molecular and pathological signatures of epithelial–mesenchymal transitions at the cancer invasion front. Histochem Cell Biol. 2008;130:481–94. PubMed: 18648847.CrossRefPubMedPubMedCentralGoogle Scholar
Merikallio H, Kaarteenaho R, Pääkkö P, Lehtonen S, Hirvikoski P, Mäkitaro R, et al. Zeb1 and twist are more commonly expressed in metastatic than primary lung tumours and show inverse associations with claudins. J Clin Pathol. 2011;64:136–40. PubMed: 21131312.CrossRefPubMedGoogle Scholar
Lee KW, Lee NK, Kim JH, Kang MS, Yoo HY, Kim HH, et al. Twist1 causes the transcriptional repression of claudin-4 with prognostic significance in esophageal cancer. Biochem Biophys Res Commun. 2012;423:454–60. PubMed: 22668877.CrossRefPubMedGoogle Scholar
Jung JH, Jung CK, Choi HJ. Diagnostic utility of expression of claudins in non-small cell lung cancer: different expression profiles in squamous cell carcinomas and adenocarcinomas. Pathol Res Pract. 2009;205:409–16. PubMed: 19231096.CrossRefPubMedGoogle Scholar
Romanov V, Whyard TC, Waltzer WC, Gabig TG. A claudin 3 and claudin 4-targeted Clostridium perfringens protoxin is selectively cytotoxic to PSA-producing prostate cancer cells. Cancer Lett. 2014;351:260–4. PubMed: 24952257.CrossRefPubMedGoogle Scholar
Yao Q, Zheng QM, Wen JF, Lü T, Wei MQ, Dai SZ. Target-specific cytotoxic activity of recombinant fusion toxin C-CPE-ETA’ against CLDN-3,4-overexpressing ovarian cancer cells. Zhonghua Zhong Liu Za Zhi. 2010;32:897–902. PubMed: 21223796.PubMedGoogle Scholar
Ersoz S, Mungan S, Cobanoglu U, Turgutalp H, Ozoran Y. Prognostic importance of claudin-1 and claudin-4 expression in colon carcinomas. Pathol Res Pract. 2011;207:285–9. PubMed: 21493012.CrossRefPubMedGoogle Scholar
Merikallio H, Pääkkö P, Harju T, Soini Y. Claudins 10 and 18 are predominantly expressed in lung adenocarcinomas and in tumors of nonsmokers. Int J Clin Exp Pathol. 2011;4:667–73. PubMed: 22076167.PubMedPubMedCentralGoogle Scholar
Prudkin L, Liu DD, Ozburn NC, Sun M, Behrens C, Tang X, et al. Epithelial-to-mesenchymal transition in the development and progression of adenocarcinoma and squamous cell carcinoma of the lung. Mod Pathol. 2009;22:668–78. PubMed: 19270647.CrossRefPubMedPubMedCentralGoogle Scholar
Hirata T, Fukuse T, Naiki H, Wada H. Expression of E-cadherin and lymph node metastasis in resected non-small-cell lung cancer. Clin Lung Cancer. 2001;3:134–40. PubMed: 14659029.CrossRefPubMedGoogle Scholar
Hommura F, Furuuchi K, Yamazaki K, Ogura S, Kinoshita I, Shimizu M, et al. Increased expression of beta-catenin predicts better prognosis in nonsmall cell lung carcinomas. Cancer. 2002;94:752–8. PubMed: 11857309.CrossRefPubMedGoogle Scholar
Zhang Y, Han Y, Zheng R, Yu J-H, Miao Y, Wang L, et al. Expression of Frat1 correlates with expression of β-catenin and is associated with a poor clinical outcome in human SCC and AC. Tumour Biol. 2012;33:1437–44. PubMed: 22528942.CrossRefPubMedGoogle Scholar
Pirinen RT, Hirvikoski P, Johansson RT, Hollmén S, Kosma VM. Reduced expression of alpha-catenin, beta-catenin, and gamma-catenin is associated with high cell proliferative activity and poor differentiation in non-small cell lung cancer. J Clin Pathol. 2001;54:391–5. PubMed: 11328840.CrossRefPubMedPubMedCentralGoogle Scholar
Zhang H, Liu J, Yue D, Gao L, Wang D, Zhang H, et al. Clinical significance of E-cadherin, β-catenin, vimentin and S100A4 expression in completely resected squamous cell lung carcinoma. J Clin Pathol. 2013;66:937–45. PubMed: 23853314.CrossRefPubMedGoogle Scholar
Kim SH, Kim JM, Shin M, Kim CW, Huang SM, Kang DW, et al. Correlation of epithelial-mesenchymal transition markers with clinicopathologic parameters in adenocarcinomas and squamous cell carcinoma of the lung. Histol Histopathol. 2012;27:581–91. PubMed: 22419022.PubMedGoogle Scholar
Peinado H, Olmeda D, Cano A. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer. 2007;7:415–28. PubMed: 17508028.CrossRefPubMedGoogle Scholar
Soini Y, Tuhkanen H, Sironen R, Virtanen I, Kataja V, Auvinen P, et al. Transcription factors zeb1, twist and snai1 in breast carcinoma. BMC Cancer. 2011;11:73. PubMed: 21324165.CrossRefPubMedPubMedCentralGoogle Scholar