Issue
Nuclear entrapment of p33ING1b by inhibition of exportin-1: A trigger of apoptosis in head and neck squamous cell cancer
Corresponding Author(s) : Sibel í–zdaş
Cellular and Molecular Biology,
Vol. 64 No. 5: Issue 5
Abstract
The effect of deregulation of nuclear export mediated by exportin-1, with consequent cellular mislocalization of p33ING1b, a member of the tumor suppressor gene family, has not been previously investigated in head and neck squamous cell cancer (HNSCC). We evaluated the effect of reversing cytoplasmic p33ING1b localization through inhibition of exportin-1 by leptomycin B (LMB) and the effect of nuclear entrapment of p33ING1b on molecular alterations in primary and metastatic HNSCC lines. The expression and location of exportin-1 and p33ING1b were analyzed by a quantitative real"‘time reverse transcription polymerase chain reaction PCR (qRT-PCR), a Western blot, and immunostaining. Cell proliferation and migration assays were conducted to determine the effect of exportin-1 inhibition on the cell lines. Exportin-1 was overexpressed in metastatic HNSCC, whereas p33ING1b was poorly expressed. Exportin-1 inhibition induced nuclear entrapment and upregulation of p33ING1b, extensive apoptosis, and growth arrest. It also suppressed cell migration. Cytoplasmic p33ING1b-mediated regulation of cell growth and nuclear entrapment of p33ING1b via inhibition of exportin-1 may be a key mechanism for inducing HNSCC apoptosis.
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- Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin 2014;64:9-29.
- Richey LM, Shores CG, George J, Lee S, Couch MJ, Sutton DK, et al. The effectiveness of salvage surgery after the failure of primary concomitant chemoradiation in head and neck cancer. Otolaryngol Head Neck Surg 2007;136:98-103.
- Turner JG, Sullivan DM. CRM1-mediated nuclear export of proteins and drug resistance in cancer. Curr Med Chem 2008;15(26):2648-2655.
- Soliman MA, and Riabowol K. After a decade of study-ING, a PHD for a versatile family of proteins. Trends Biochem Sci 2007;32:509-519.
- Gong W, Suzuki K, Russell M, Riabowol K. Function of the ING family of PHD proteins in cancer. Int J Biochem Cell Biol 2005;37(5):1054-1065.
- Li XH, Noguchi A, Nishida T, Takahashi H, Zheng Y, Yang XH, et al. Cytoplasmic expression of p33ING1b is correlated with tumorigenesis and progression of head and neck squamous cell carcinoma. Histol Histopathol 2011;26(5):597-607.
- Zhang JT, Wang DW, Li QX, Zhu ZL, Wang MW, Cui DS, et al. Nuclear to cytoplasmic shift of p33(ING1b) protein from normal oral mucosa to oral squamous cell carcinoma in relation to clinicopathological variables. J Cancer Res Clin Oncol 2008;134:421-426.
- Gong W, Russell M, Suzuki K, Riabowol K. Subcellular targeting of p33ING1b by phosphorylation-dependent 14-3-3 binding regulates p21WAF1 expression. Mol Cell Biol 2006;26(8):2947-2954.
- Fornerod M, Ohno M, Yoshida M, Mattaj IW. CRM1 is an export receptor for leucine-rich nuclear export signals. Cell 90(6):1051-1060, 1997.
- Fabbro M, Henderson, BR. Regulation of tumor suppressors by nuclear-cytoplasmic shuttling. Exp Cell Res 2003;282(2):59-69.
- Fukuda M, Asano S, Nakamura T, Adachi M, Yoshida M, Yanagida M, et al. CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature 1997;390:308-311.
- van der Watt PJ, Maske CP, Hendricks DT, Parker MI, Denny L, Govender D, et al. The Karyopherin proteins, Crm1 and Karyopherin beta1, are overexpressed in cervical cancer and are critical for cancer cell survival and proliferation. Int J Cancer 2009;124(8):1829-1840.
- Noske A, Weichert W, Niesporek S, Roske A, Buckendahl AC, Koch I, et al. Expression of the nuclear export protein chromosomal region maintenance/exportin 1/Xpo1 is a prognostic factor in human ovarian cancer. Cancer 2008;112(8): 1733-1743.
- Yao Y, Dong Y, Lin F, Zhao H, Shen Z, Chen P, et al. The expression of CRM1 is associated with prognosis in human osteosarcoma. Oncol Rep 2009;21(1):229-235.
- Huang WY, Yue L, Qiu WS, Wang LW, Zhou XH, Sun YJ. Prognostic value of CRM1 in pancreas cancer. Clin Invest Med 2009;32:E315.
- Yang X, Cheng L, Yao L, Ren H, Zhang S, Min X, et al. Involvement of chromosome region maintenance 1 (CRM1) in the formation and progression of esophageal squamous cell carcinoma. Med Oncol 2014;31(9):155.
- van der Watt PJ, Zemanay W, Govender D, Hendricks DT, Parker MI, Leaner VD. Elevated expression of the nuclear export protein, Crm1 (exportin 1), associates with human oesophageal squamous cell carcinoma. Oncol Rep 2014;32(2):730-738.
- Shen A, Wang Y, Zhao Y, Zou L, Sun L, Cheng C. Expression of CRM1 in human gliomas and its significance in p27 expression and clinical prognosis. Neurosurgery 2009;65:153-159.
- Zhou F, Qiu W, Yao R, Xiang J, Sun X, Liu S, et al. CRM1 is a novel independent prognostic factor for the poor prognosis of gastric carcinomas. Med Oncol 2013;30:726.
- Tai YT, Landesman Y, Acharya C, Calle Y, Zhong MY, Cea M, et al. CRM1 inhibition induces tumor cell cytotoxicity and impairs osteoclastogenesis in multiple myeloma: molecular mechanisms and therapeutic implications. Leukemia 2014;28:155-165.
- Newlands ES, Rustin GJ, Brampton MH. Phase I trial of elactocin. Br J Cancer 1996;74:648-649.
- Takebayashi S, Hickson A, Ogawa T, Jung KY, Mineta H, Ueda Y, et al. Loss of chromosome arm 18q with tumor progression in head and neck squamous cancer. Gene Chromosome Canc 2004;41:145-154.
- í–zdaş S, í–zdaş T. Crm1 knockdown by specific small interfering RNA reduces cell proliferation and induces apoptosis in head and neck cancer cell lines. Turk J Biol 2018;42:1-12.
- Gokturk D, Kelebek H, Ceylan S, Yilmaz DM. The Effect of Ascorbic Acid over the Etoposide- and Temozolomide-Mediated Cytotoxicity in Glioblastoma Cell Culture: A Molecular Study. Turk Neurosurg 2018;28(1):13-18.
- í–zdaş S, í–zdaş T. Differentially expressed Secretoglobin 1C1 gene in nasal polyposis. Cell Mol Biol (Noisy-le-grand) 2018;64(1):97-102.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001;25:402-408.
- Yadav A, Kumar B, Datta J, Teknos TN, Kumar P. IL-6 Promotes Head and Neck Tumor Metastasis by Inducing Epithelial-Mesenchymal Transition via the JAK-STAT3-SNAIL Signaling Pathway. Mol Cancer Res 2011;9:1658-1667.
- Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol Annu Rev 2005;11:127-152.
- Russell MW, Soliman MA, Schriemer D and Riabowol K. ING1 protein targeting to the nucleus by karyopherins is necessary for activation of p21. Biochem Bioph Res Commun 2008;374:490-495.
- Zhu JJ, Li FB, Zhou JM, Liu ZC, Zhu XF, Liao WM. The tumor suppressor p33ING1b enhances taxol-induced apoptosis by p53-dependent pathway in human osteosarcoma U2OS cells. Cancer Biol Ther 2005;4(1):39-47.
- Easty D, Easty G, Carter R, Monaghan P, Butler L. Ten human carcinoma cell lines derived from squamous carcinomas of the head and neck. Br J Cancer 1981;43:772-785.
- Mutka SC, Yang WQ, Dong SD, Ward SL, Craig DA, Timmermans PB, et al. Identification of nuclear export inhibitors with potent anticancer activity in vivo. Cancer Res 2009;69(2):510-517.
- Kau TR, Way JC, Silver PA. Nuclear transport and cancer: from mechanism to intervention. Nat Rev Cancer 2004;4:106-117.
- Kuusisto HV, Wagstaff KM, Alvisi G, Roth DM, Jans DA. Global enhancement of nuclear localization-dependent nuclear transport in transformed cells. FASEB J 2011;26:1181-1193.
References
Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin 2014;64:9-29.
Richey LM, Shores CG, George J, Lee S, Couch MJ, Sutton DK, et al. The effectiveness of salvage surgery after the failure of primary concomitant chemoradiation in head and neck cancer. Otolaryngol Head Neck Surg 2007;136:98-103.
Turner JG, Sullivan DM. CRM1-mediated nuclear export of proteins and drug resistance in cancer. Curr Med Chem 2008;15(26):2648-2655.
Soliman MA, and Riabowol K. After a decade of study-ING, a PHD for a versatile family of proteins. Trends Biochem Sci 2007;32:509-519.
Gong W, Suzuki K, Russell M, Riabowol K. Function of the ING family of PHD proteins in cancer. Int J Biochem Cell Biol 2005;37(5):1054-1065.
Li XH, Noguchi A, Nishida T, Takahashi H, Zheng Y, Yang XH, et al. Cytoplasmic expression of p33ING1b is correlated with tumorigenesis and progression of head and neck squamous cell carcinoma. Histol Histopathol 2011;26(5):597-607.
Zhang JT, Wang DW, Li QX, Zhu ZL, Wang MW, Cui DS, et al. Nuclear to cytoplasmic shift of p33(ING1b) protein from normal oral mucosa to oral squamous cell carcinoma in relation to clinicopathological variables. J Cancer Res Clin Oncol 2008;134:421-426.
Gong W, Russell M, Suzuki K, Riabowol K. Subcellular targeting of p33ING1b by phosphorylation-dependent 14-3-3 binding regulates p21WAF1 expression. Mol Cell Biol 2006;26(8):2947-2954.
Fornerod M, Ohno M, Yoshida M, Mattaj IW. CRM1 is an export receptor for leucine-rich nuclear export signals. Cell 90(6):1051-1060, 1997.
Fabbro M, Henderson, BR. Regulation of tumor suppressors by nuclear-cytoplasmic shuttling. Exp Cell Res 2003;282(2):59-69.
Fukuda M, Asano S, Nakamura T, Adachi M, Yoshida M, Yanagida M, et al. CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature 1997;390:308-311.
van der Watt PJ, Maske CP, Hendricks DT, Parker MI, Denny L, Govender D, et al. The Karyopherin proteins, Crm1 and Karyopherin beta1, are overexpressed in cervical cancer and are critical for cancer cell survival and proliferation. Int J Cancer 2009;124(8):1829-1840.
Noske A, Weichert W, Niesporek S, Roske A, Buckendahl AC, Koch I, et al. Expression of the nuclear export protein chromosomal region maintenance/exportin 1/Xpo1 is a prognostic factor in human ovarian cancer. Cancer 2008;112(8): 1733-1743.
Yao Y, Dong Y, Lin F, Zhao H, Shen Z, Chen P, et al. The expression of CRM1 is associated with prognosis in human osteosarcoma. Oncol Rep 2009;21(1):229-235.
Huang WY, Yue L, Qiu WS, Wang LW, Zhou XH, Sun YJ. Prognostic value of CRM1 in pancreas cancer. Clin Invest Med 2009;32:E315.
Yang X, Cheng L, Yao L, Ren H, Zhang S, Min X, et al. Involvement of chromosome region maintenance 1 (CRM1) in the formation and progression of esophageal squamous cell carcinoma. Med Oncol 2014;31(9):155.
van der Watt PJ, Zemanay W, Govender D, Hendricks DT, Parker MI, Leaner VD. Elevated expression of the nuclear export protein, Crm1 (exportin 1), associates with human oesophageal squamous cell carcinoma. Oncol Rep 2014;32(2):730-738.
Shen A, Wang Y, Zhao Y, Zou L, Sun L, Cheng C. Expression of CRM1 in human gliomas and its significance in p27 expression and clinical prognosis. Neurosurgery 2009;65:153-159.
Zhou F, Qiu W, Yao R, Xiang J, Sun X, Liu S, et al. CRM1 is a novel independent prognostic factor for the poor prognosis of gastric carcinomas. Med Oncol 2013;30:726.
Tai YT, Landesman Y, Acharya C, Calle Y, Zhong MY, Cea M, et al. CRM1 inhibition induces tumor cell cytotoxicity and impairs osteoclastogenesis in multiple myeloma: molecular mechanisms and therapeutic implications. Leukemia 2014;28:155-165.
Newlands ES, Rustin GJ, Brampton MH. Phase I trial of elactocin. Br J Cancer 1996;74:648-649.
Takebayashi S, Hickson A, Ogawa T, Jung KY, Mineta H, Ueda Y, et al. Loss of chromosome arm 18q with tumor progression in head and neck squamous cancer. Gene Chromosome Canc 2004;41:145-154.
í–zdaş S, í–zdaş T. Crm1 knockdown by specific small interfering RNA reduces cell proliferation and induces apoptosis in head and neck cancer cell lines. Turk J Biol 2018;42:1-12.
Gokturk D, Kelebek H, Ceylan S, Yilmaz DM. The Effect of Ascorbic Acid over the Etoposide- and Temozolomide-Mediated Cytotoxicity in Glioblastoma Cell Culture: A Molecular Study. Turk Neurosurg 2018;28(1):13-18.
í–zdaş S, í–zdaş T. Differentially expressed Secretoglobin 1C1 gene in nasal polyposis. Cell Mol Biol (Noisy-le-grand) 2018;64(1):97-102.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001;25:402-408.
Yadav A, Kumar B, Datta J, Teknos TN, Kumar P. IL-6 Promotes Head and Neck Tumor Metastasis by Inducing Epithelial-Mesenchymal Transition via the JAK-STAT3-SNAIL Signaling Pathway. Mol Cancer Res 2011;9:1658-1667.
Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol Annu Rev 2005;11:127-152.
Russell MW, Soliman MA, Schriemer D and Riabowol K. ING1 protein targeting to the nucleus by karyopherins is necessary for activation of p21. Biochem Bioph Res Commun 2008;374:490-495.
Zhu JJ, Li FB, Zhou JM, Liu ZC, Zhu XF, Liao WM. The tumor suppressor p33ING1b enhances taxol-induced apoptosis by p53-dependent pathway in human osteosarcoma U2OS cells. Cancer Biol Ther 2005;4(1):39-47.
Easty D, Easty G, Carter R, Monaghan P, Butler L. Ten human carcinoma cell lines derived from squamous carcinomas of the head and neck. Br J Cancer 1981;43:772-785.
Mutka SC, Yang WQ, Dong SD, Ward SL, Craig DA, Timmermans PB, et al. Identification of nuclear export inhibitors with potent anticancer activity in vivo. Cancer Res 2009;69(2):510-517.
Kau TR, Way JC, Silver PA. Nuclear transport and cancer: from mechanism to intervention. Nat Rev Cancer 2004;4:106-117.
Kuusisto HV, Wagstaff KM, Alvisi G, Roth DM, Jans DA. Global enhancement of nuclear localization-dependent nuclear transport in transformed cells. FASEB J 2011;26:1181-1193.