KPNA2 is a promising biomarker candidate for esophageal squamous cell carcinoma and correlates with cell proliferation

Ma,Shouzhi; Zhao,Xiaohang

doi:10.3892/or.2014.3381

KPNA2 is a promising biomarker candidate for esophageal squamous cell carcinoma and correlates with cell proliferation

Authors:
- Shouzhi Ma
- Xiaohang Zhao
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Affiliations: State Key Laboratory of Molecular Oncology, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, P.R. China
Published online on: August 4, 2014 https://doi.org/10.3892/or.2014.3381
Pages: 1631-1637

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Abstract

Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignant cancers worldwide, with a poor 5-year prognosis. Karyopherin α 2 (KPNA2) is a nuclear membrane protein that mediates nucleus-to-cytoplasm shuttling. Its expression is elevated in multiple forms of cancer, and it can be secreted into the serum. However, the concentration of KPNA2 in serum from ESCC patients and the role of KPNA2 in ESCC cells remains unclear. The aim of the present study was to determine the concentration of KPNA2 in serum from ESCC patients and to investigate the effect of KPNA2 silencing on ESCC cell proliferation. KPNA2 protein expression was detected at the tissue level by immunohistochemistry, in cell lines by western blotting and at the serum level by enzyme linked immunosorbent assay (ELISA). Cell proliferation was determined by cell growth curve and colony formation assay. Stages of the cell cycle were analyzed by flow cytometry. The effect of KPNA2 knockdown on E2F1 translocation was determined by subcellular fractionation. KPNA2 was overexpressed in both ESCC tissues and cell lines compared with controls. The concentration of KPNA2 in serum from ESCC patients was significantly higher than that from healthy controls. The AUC was determined to be 0.804. The sensitivity and specificity of the assay were 76.7 and 75.0%, respectively. To determine the significance of KPNA2 function, small interfering RNA (siRNA) against KPNA2 was used to knock down KPNA2 levels in the ESCC using siRNA in the Kyse510 cell line. KPNA2 siRNA inhibited Kyse510 cell proliferation and colony formation ability and induced a G2/M phase arrest. The nuclear translocation of E2F1 was also reduced in siRNA-treated Kyse510 cells. The KPNA2 protein levels were high in ESCC tumors, and siRNA against KPNA2 could inhibit the growth of ESCC cells, suggesting it may be a new potent marker and therapeutic target for ESCC.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	Wang GQ, Abnet CC, Shen Q, Lewin KJ, Sun XD, Roth MJ, Qiao YL, Mark SD, Dong ZW, Taylor PR and Dawsey SM: Histological precursors of oesophageal squamous cell carcinoma: results from a 13 year prospective follow up study in a high risk population. Gut. 54:187–192. 2005.PubMed/NCBI
3	Kelley JB, Talley AM, Spencer A, Gioeli D and Paschal BM: Karyopherin α7 (KPNA7), a divergent member of the importin α family of nuclear import receptors. BMC Cell Biol. 11:632010.
4	Stewart M: Molecular mechanism of the nuclear protein import cycle. Nat Rev Mol Cell Biol. 8:195–208. 2007. View Article : Google Scholar : PubMed/NCBI
5	Goldfarb DS, Corbett AH, Mason DA, Harreman MT and Adam SA: Importin α: a multipurpose nuclear-transport receptor. Trends Cell Biol. 14:505–514. 2004.
6	Dahl E, Kristiansen G, Gottlob K, Klaman I, Ebner E, Hinzmann B, Hermann K, Pilarsky C, Dürst M, Klinkhammer-Schalke M, Blaszyk H, Knuechel R, Hartmann A, Rosenthal A and Wild PJ: Molecular profiling of laser-microdissected matched tumor and normal breast tissue identifies karyopherin α2 as a potential novel prognostic marker in breast cancer. Clin Cancer Res. 12:3950–3960. 2006.PubMed/NCBI
7	Dankof A, Fritzsche FR, Dahl E, Pahl S, Wild P, Dietel M, Hartmann A and Kristiansen G: KPNA2 protein expression in invasive breast carcinoma and matched peritumoral ductal carcinoma in situ. Virchows Arch. 451:877–881. 2007. View Article : Google Scholar : PubMed/NCBI
8	Gluz O, Wild P, Meiler R, Diallo-Danebrock R, Ting E, Mohrmann S, Schuett G, Dahl E, Fuchs T, Herr A, Gaumann A, Frick M, Poremba C, Nitz UA and Hartmann A: Nuclear karyopherin α2 expression predicts poor survival in patients with advanced breast cancer irrespective of treatment intensity. Int J Cancer. 123:1433–1438. 2008.
9	Zheng M, Tang L, Huang L, Ding H, Liao WT, Zeng MS and Wang HY: Overexpression of karyopherin-2 in epithelial ovarian cancer and correlation with poor prognosis. Obstet Gynecol. 116:884–891. 2010. View Article : Google Scholar : PubMed/NCBI
10	Winnepenninckx V1, Lazar V, Michiels S, Dessen P, Stas M, Alonso SR, Avril MF, Ortiz Romero PL, Robert T, Balacescu O, Eggermont AM, Lenoir G, Sarasin A, Tursz T, van den Oord JJ and Spatz A: Melanoma Group of the European Organization for Research and Treatment of Cancer: Gene expression profiling of primary cutaneous melanoma and clinical outcome. J Natl Cancer Inst. 98:472–482. 2006. View Article : Google Scholar : PubMed/NCBI
11	van der Watt PJ, Maske CP, Hendricks DT, Parker MI, Denny L, Govender D, Birrer MJ and Leaner VD: The Karyopherin proteins, Crm1 and Karyopherin β1, are overexpressed in cervical cancer and are critical for cancer cell survival and proliferation. Int J Cancer. 124:1829–1840. 2009.
12	Wang CI, Wang CL, Wang CW, Chen CD, Wu CC, Liang Y, Tsai YH, Chang YS, Yu JS and Yu CJ: Importin subunit alpha-2 is identified as a potential biomarker for non-small cell lung cancer by integration of the cancer cell secretome and tissue transcriptome. Int J Cancer. 128:2364–2372. 2011. View Article : Google Scholar : PubMed/NCBI
13	Gousias K, Becker AJ, Simon M and Niehusmann P: Nuclear karyopherin a2: a novel biomarker for infiltrative astrocytomas. J Neurooncol. 109:545–553. 2012. View Article : Google Scholar : PubMed/NCBI
14	Mortezavi A, Hermanns T, Seifert HH, Baumgartner MK, Provenzano M, Sulser T, Burger M, Montani M, Ikenberg K, Hofstädter F, Hartmann A, Jaggi R, Moch H, Kristiansen G and Wild PJ: KPNA2 expression is an independent adverse predictor of biochemical recurrence after radical prostatectomy. Clin Cancer Res. 17:1111–1121. 2011. View Article : Google Scholar : PubMed/NCBI
15	Yoshitake K, Tanaka S, Mogushi K, Aihara A, Murakata A, Matsumura S, Mitsunori Y, Yasen M, Ban D, Noguchi N, Irie T, Kudo A, Nakamura N, Tanaka H and Arii S: Importin-α1 as a novel prognostic target for hepatocellular carcinoma. Ann Surg Oncol. 18:2093–2103. 2011.
16	Jensen JB1, Munksgaard PP, Sørensen CM, Fristrup N, Birkenkamp-Demtroder K, Ulhøi BP, Jensen KM, Ørntoft TF and Dyrskjøt L: High expression of karyopherin-α2 defines poor prognosis in non-muscle-invasive bladder cancer and in patients with invasive bladder cancer undergoing radical cystectomy. Eur Urol. 59:841–848. 2011.
17	Sakai M, Sohda M, Miyazaki T, Suzuki S, Sano A, Tanaka N, Inose T, Nakajima M, Kato H and Kuwano H: Significance of karyopherin-α 2 (KPNA2) expression in esophageal squamous cell carcinoma. Anticancer Res. 30:851–856. 2010.
18	Sun Y, Mi W, Cai J, Ying W, Liu F, Lu H, Qiao Y, Jia W, Bi X, Lu N, Liu S, Qian X and Zhao X: Quantitative proteomic signature of liver cancer cells: tissue transglutaminase 2 could be a novel protein candidate of human hepatocellular carcinoma. J Proteome Res. 7:3847–3859. 2008. View Article : Google Scholar
19	Xu Y, Zhou L, Huang J, Liu F, Yu J, Zhan Q, Zhang L and Zhao X: Role of Smac in determining the chemotherapeutic response of esophageal squamous cell carcinoma. Clin Cancer Res. 17:5412–5422. 2011. View Article : Google Scholar : PubMed/NCBI
20	van der Watt PJ, Ngarande E and Leaner VD: Overexpression of Kpnβ1 and Kpnα2 importin proteins in cancer derives from deregulated E2F activity. PLoS One. 6:e277232011.
21	Wang CI, Chien KY, Wang CL, Liu HP, Cheng CC, Chang YS, Yu JS and Yu CJ: Quantitative proteomics reveals regulation of karyopherin subunit alpha-2 (KPNA2) and its potential novel cargo proteins in nonsmall cell lung cancer. Mol Cell Proteomics. 11:1105–1122. 2012. View Article : Google Scholar : PubMed/NCBI
22	Ishida S, Huang E, Zuzan H, Spang R, Leone G, West M and Nevins JR: Role for E2F in control of both DNA replication and mitotic functions as revealed from DNA microarray analysis. Mol Cell Biol. 21:4684–4699. 2001. View Article : Google Scholar : PubMed/NCBI
23	Zhu W, Giangrande PH and Nevins JR: E2Fs link the control of G1/S and G2/M transcription. EMBO J. 23:4615–4626. 2004. View Article : Google Scholar : PubMed/NCBI
24	Dong QM, Zhang JQ, Li Q, Bracher JC, Hendricks DT and Zhao XH: Clinical significance of serum expression of GROβ in esophageal squamous cell carcinoma. World J Gastroenterol. 17:2658–2662. 2011.
25	Quensel C, Friedrich B, Sommer T, Hartmann E and Kohler M: In vivo analysis of importin α proteins reveals cellular proliferation inhibition and substrate specificity. Mol Cell Biol. 24:10246–10255. 2004.
26	Huang L, Wang HY, Li JD, Wang JH, Zhou Y, Luo RZ, Yun JP, Zhang Y, Jia WH and Zheng M: KPNA2 promotes cell proliferation and tumorigenicity in epithelial ovarian carcinoma through upregulation of c-Myc and downregulation of FOXO3a. Cell Death Dis. 4:e7452013. View Article : Google Scholar : PubMed/NCBI
27	Ren B, Cam H, Takahashi Y, Volkert T, Terragni J, Young RA and Dynlacht BD: E2F integrates cell cycle progression with DNA repair, replication, and G₂/M checkpoints. Genes Dev. 16:245–256. 2002. View Article : Google Scholar : PubMed/NCBI
28	Zannini L, Lecis D, Lisanti S, Benetti R, Buscemi G, Schneider C and Delia D: Karyopherin-α2 protein interacts with Chk2 and contributes to its nuclear import. J Biol Chem. 278:42346–42351. 2003.
29	Narod SA and Foulkes WD: BRCA1 and BRCA2: 1994 and beyond. Nat Rev Cancer. 4:665–676. 2004. View Article : Google Scholar
30	Tseng SF, Chang CY, Wu KJ and Teng SC: Importin KPNA2 is required for proper nuclear localization and multiple functions of NBS1. J Biol Chem. 280:39594–39600. 2005. View Article : Google Scholar : PubMed/NCBI
31	Sandrock K, Bielek H, Schradi K, Schmidt G and Klugbauer N: The nuclear import of the small GTPase Rac1 is mediated by the direct interaction with karyopherin α2. Traffic. 11:198–209. 2010.PubMed/NCBI
32	Ebihara Y, Miyamoto M, Shichinohe T, Kawarada Y, Cho Y, Fukunaga A, Murakami S, Uehara H, Kaneko H, Hashimoto H, Murakami Y, Itoh T, Okushiba S, Kondo S and Katoh H: Overexpression of E2F-1 in esophageal squamous cell carcinoma correlates with tumor progression. Dis Esophagus. 17:150–154. 2004. View Article : Google Scholar : PubMed/NCBI