Heterogeneous nuclear ribonucleoprotein K is overexpressed and contributes to radioresistance irrespective of HPV status in head and neck squamous cell carcinoma

Kähler,Justus; Hafner,Susanne; Popp,Tanja; Hermann,Cornelius; Rump,Alexis; Port,Matthias; Steinestel,Konrad; Eder,Stefan

doi:10.3892/ijmm.2020.4718

Heterogeneous nuclear ribonucleoprotein K is overexpressed and contributes to radioresistance irrespective of HPV status in head and neck squamous cell carcinoma

Authors:
- Justus Kähler
- Susanne Hafner
- Tanja Popp
- Cornelius Hermann
- Alexis Rump
- Matthias Port
- Konrad Steinestel
- Stefan Eder
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Affiliations: Bundeswehr Institute of Radiobiology, D‑80937 Munich, Germany, Institute of Pharmacology of Natural Products and Clinical Pharmacology, University of Ulm, D‑89081 Ulm, Germany, Institute of Pathology and Molecular Pathology, Bundeswehrkrankenhaus Ulm, D‑89081 Ulm, Germany
Published online on: September 2, 2020 https://doi.org/10.3892/ijmm.2020.4718
Pages: 1733-1742
Copyright: © Kähler et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Radiotherapy is a major treatment option for head and neck squamous cell carcinoma (HNSCC). However, the success of radiotherapy is limited by tumor cell resistance to ionizing radiation (IR). Clinical studies have demonstrated an overall improved prognosis and higher susceptibility to radiotherapy of high‑risk human papillomavirus (HPV)‑associated HNSCC compared with classic HNSCC, as well as worse overall survival for male HNSCC patients. Overexpression of heterogeneous nuclear ribonucleoprotein (hnRNP) K has been associated with resistance to radiotherapy in melanoma and colorectal carcinoma. The aim of the present study was to analyze the impact of hnRNP K expression on the aggressiveness and radioresistance of HNSCC with respect to patient sex and HPV status. Immunohistochemical staining of HNSCC tissue specimens revealed elevated hnRNP K levels compared with those in the non‑neoplastic epithelium. Cytoplasmic hnRNP K accumulation was associated with advanced tumor stage and male sex. Exposure of HNSCC cells to IR was followed by rapid upregulation of hnRNP K at the protein level, along with re‑localization from the tumor cell nucleus to the cytoplasm. siRNA‑based knockdown of hnRNP K induced apoptosis and abolished tumor formation after xenotransplantation of HNSCC cells onto the chick egg chorioallantoic membrane (CAM). The observed effects were independent of the respective HPV status of the cell lines. These results indicated a tumorigenic and anti‑apoptotic role of hnRNP K in HNSCC, which appeared to be enhanced in male patients and contributed to the radioresistance of these tumors. However, the radioprotective effects of hnRNP K were found to be independent of the tumor's HPV status.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
3	Franceschi S, Talamini R, Barra S, Baron AE, Negri E, Bidoli E, Serraino D and La Vecchia C: Smoking and drinking in relation to cancers of the oral cavity, pharynx, larynx, and esophagus in northern Italy. Cancer Res. 50:6502–6507. 1990.PubMed/NCBI
4	Ang KK and Sturgis EM: Human papillomavirus as a marker of the natural history and response to therapy of head and neck squamous cell carcinoma. Semin Radiat Oncol. 22:128–142. 2012. View Article : Google Scholar : PubMed/NCBI
5	Chaturvedi AK: Epidemiology and clinical aspects of HPV in head and neck cancers. Head Neck Pathol. 6(Suppl 1): S16–S24. 2012. View Article : Google Scholar : PubMed/NCBI
6	Chaturvedi AK, Engels EA, Pfeiffer RM, Hernandez BY, Xiao W, Kim E, Jiang B, Goodman MT, Sibug-Saber M, Cozen W, et al: Human papillomavirus and rising oropharyngeal cancer incidence in the United States. J Clin Oncol. 29:4294–4301. 2011. View Article : Google Scholar : PubMed/NCBI
7	Brouwer AF, Eisenberg MC and Meza R: Age effects and temporal trends in HPV-Related and HPV-Unrelated oral cancer in the United States: A multistage carcinogenesis modeling analysis. PLoS One. 11:e01510982016. View Article : Google Scholar : PubMed/NCBI
8	Fakhry C, Westra WH, Li S, Cmelak A, Ridge JA, Pinto H, Forastiere A and Gillison ML: Improved survival of patients with human papillomavirus-positive head and neck squamous cell carcinoma in a prospective clinical trial. J Natl Cancer Inst. 100:261–269. 2008. View Article : Google Scholar : PubMed/NCBI
9	Liu C, Mann D, Sinha UK and Kokot NC: The molecular mechanisms of increased radiosensitivity of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC): An extensive review. J Otolaryngol Head Neck Surg. 47:592018. View Article : Google Scholar : PubMed/NCBI
10	Moody CA and Laimins LA: Human papillomavirus oncoproteins: Pathways to transformation. Nat Rev Cancer. 10:550–560. 2010. View Article : Google Scholar : PubMed/NCBI
11	Barboro P, Ferrari N and Balbi C: Emerging roles of heterogeneous nuclear ribonucleoprotein K (hnRNP K) in cancer progression. Cancer Lett. 352:152–159. 2014. View Article : Google Scholar : PubMed/NCBI
12	Gallardo M, Hornbaker MJ, Zhang X, Hu P, Bueso-Ramos C and Post SM: Aberrant hnRNP K expression: All roads lead to cancer. Cell Cycle. 15:1552–1557. 2016. View Article : Google Scholar : PubMed/NCBI
13	Bomsztyk K, Denisenko O and Ostrowski J: hnRNP K: One protein multiple processes. Bioessays. 26:629–638. 2004. View Article : Google Scholar : PubMed/NCBI
14	Carpenter B, McKay M, Dundas SR, Lawrie LC, Telfer C and Murray GI: Heterogeneous nuclear ribonucleoprotein K is over expressed, aberrantly localised and is associated with poor prognosis in colorectal cancer. Br J Cancer. 95:921–927. 2006. View Article : Google Scholar : PubMed/NCBI
15	Chen LC, Chung IC, Hsueh C, Tsang NM, Chi LM, Liang Y, Chen CC, Wang LJ and Chang YS: The antiapoptotic protein, FLIP, is regulated by heterogeneous nuclear ribonucleoprotein K and correlates with poor overall survival of nasopharyngeal carcinoma patients. Cell Death Differ. 17:1463–1473. 2010. View Article : Google Scholar : PubMed/NCBI
16	Wu CS, Chang KP, Chen LC, Chen CC, Liang Y, Hseuh C and Chang YS: Heterogeneous ribonucleoprotein K and thymidine phosphorylase are independent prognostic and therapeutic markers for oral squamous cell carcinoma. Oral Oncol. 48:516–522. 2012. View Article : Google Scholar : PubMed/NCBI
17	Chen X, Gu P, Xie R, Han J, Liu H, Wang B, Xie W, Xie W, Zhong G, Chen C, et al: Heterogeneous nuclear ribonucleoprotein K is associated with poor prognosis and regulates proliferation and apoptosis in bladder cancer. J Cell Mol Med. 21:1266–1279. 2017. View Article : Google Scholar :
18	Eder S, Arndt A, Lamkowski A, Daskalaki W, Rump A, Priller M, Genze F, Wardelmann E, Port M and Steinestel K: Baseline MAPK signaling activity confers intrinsic radioresistance to KRAS-mutant colorectal carcinoma cells by rapid upregulation of heterogeneous nuclear ribonucleoprotein K (hnRNP K). Cancer Lett. 385:160–167. 2017. View Article : Google Scholar
19	Matta A, Tripathi SC, DeSouza LV, Grigull J, Kaur J, Chauhan SS, Srivastava A, Thakar A, Shukla NK, Duggal R, et al: Heterogeneous ribonucleoprotein K is a marker of oral leukoplakia and correlates with poor prognosis of squamous cell carcinoma. Int J Cancer. 125:1398–1406. 2009. View Article : Google Scholar : PubMed/NCBI
20	Moumen A, Masterson P, O'Connor MJ and Jackson SP: hnRNP K: An HDM2 target and transcriptional coactivator of p53 in response to DNA damage. Cell. 123:1065–1078. 2005. View Article : Google Scholar : PubMed/NCBI
21	Haley B, Paunesku T, Protic M and Woloschak GE: Response of heterogeneous ribonuclear proteins (hnRNP) to ionising radiation and their involvement in DNA damage repair. Int J Radiat Biol. 85:643–655. 2009. View Article : Google Scholar : PubMed/NCBI
22	Wiesmann N, Strozynski J, Beck C, Zimmermann N, Mendler S, Gieringer R, Schmidtmann I and Brieger J: Knockdown of hnRNPK leads to increased DNA damage after irradiation and reduces survival of tumor cells. Carcinogenesis. 38:321–328. 2017. View Article : Google Scholar : PubMed/NCBI
23	Eder S, Lamkowski A, Priller M, Port M and Steinestel K: Radiosensitization and downregulation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) upon inhibition of mitogen/extracellular signal-regulated kinase (MEK) in malignant melanoma cells. Oncotarget. 6:17178–17191. 2015. View Article : Google Scholar : PubMed/NCBI
24	Strozynski J, Heim J, Bunbanjerdsuk S, Wiesmann N, Zografidou L, Becker SK, Meierl AM, Gouveris H, Lüddens H, Grus F and Brieger J: Proteomic identification of the heterogeneous nuclear ribonucleoprotein K as irradiation responsive protein related to migration. J Proteomics. 113:154–161. 2015. View Article : Google Scholar
25	Steinestel K, Lennerz JK, Eder S, Kraft K and Arndt A: Invasion pattern and histologic features of tumor aggressiveness correlate with MMR protein expression, but are independent of activating KRAS and BRAF mutations in CRC. Virchows Arch. 465:155–163. 2014. View Article : Google Scholar : PubMed/NCBI
26	Liebau S, Steinestel J, Linta L, Kleger A, Storch A, Schoen M, Steinestel K, Proepper C, Bockmann J, Schmeisser MJ and Boeckers TM: An SK3 channel/nWASP/Abi-1 complex is involved in early neurogenesis. PLoS One. 6:e181482011. View Article : Google Scholar : PubMed/NCBI
27	Kuan SL, Fischer S, Hafner S, Wang T, Syrovets T, Liu W, Tokura Y, Ng DYW, Riegger A, Förtsch C, et al: Boosting antitumor drug efficacy with chemically engineered multidomain proteins. Adv Sci (Weinh). 5:17010362018. View Article : Google Scholar
28	Zuo Z, Syrovets T, Wu Y, Hafner S, Vernikouskaya I, Liu W, Ma G, Weil T, Simmet T and Rasche V: The CAM cancer xenograft as a model for initial evaluation of MR labelled compounds. Sci Rep. 7:466902017. View Article : Google Scholar : PubMed/NCBI
29	Tomayko MM and Reynolds CP: Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol. 24:148–154. 1989. View Article : Google Scholar : PubMed/NCBI
30	Fakhry C, Westra WH, Wang SJ, van Zante A, Zhang Y, Rettig E, Yin LX, Ryan WR, Ha PK, Wentz A, et al: The prognostic role of sex, race, and human papillomavirus in oropharyngeal and nonoropharyngeal head and neck squamous cell cancer. Cancer. 123:1566–1575. 2017. View Article : Google Scholar : PubMed/NCBI
31	Worsham MJ, Stephen JK, Chen KM, Mahan M, Schweitzer V, Havard S and Divine G: Improved survival with HPV among African Americans with oropharyngeal cancer. Clin Cancer Res. 19:2486–2492. 2013. View Article : Google Scholar : PubMed/NCBI
32	Roychoudhury P and Chaudhuri K: Evidence for heterogeneous nuclear ribonucleoprotein K overexpression in oral squamous cell carcinoma. Br J Cancer. 97:574–576. 2007. View Article : Google Scholar : PubMed/NCBI
33	Gao R, Yu Y, Inoue A, Widodo N, Kaul SC and Wadhwa R: Heterogeneous nuclear ribonucleoprotein K (hnRNP-K) promotes tumor metastasis by induction of genes involved in extracellular matrix, cell movement, and angiogenesis. J Biol Chem. 288:15046–15056. 2013. View Article : Google Scholar : PubMed/NCBI
34	Chung IC, Chen LC, Chung AK, Chao M, Huang HY, Hsueh C, Tsang NM, Chang KP, Liang Y, Li HP and Chang YS: Matrix metalloproteinase 12 is induced by heterogeneous nuclear ribonucleoprotein K and promotes migration and invasion in nasopharyngeal carcinoma. BMC Cancer. 14:3482014. View Article : Google Scholar : PubMed/NCBI
35	Ang KK, Zhang Q, Rosenthal DI, Nguyen-Tan PF, Sherman EJ, Weber RS, Galvin JM, Bonner JA, Harris J, El-Naggar AK, et al: Randomized phase III trial of concurrent accelerated radiation plus cisplatin with or without cetuximab for stage III to IV head and neck carcinoma: RTOG 0522. J Clin Oncol. 32:2940–2950. 2014. View Article : Google Scholar : PubMed/NCBI
36	Gao X, Feng J, He Y, Xu F, Fan X, Huang W, Xiong H, Liu Q, Liu W, Liu X, et al: hnRNPK inhibits GSK3β Ser9 phosphorylation, thereby stabilizing c-FLIP and contributes to TRAIL resistance in H1299 lung adenocarcinoma cells. Sci Rep. 6:229992016. View Article : Google Scholar
37	Yang JH, Chiou YY, Fu SL, Shih IY, Weng TH, Lin WJ and Lin CH: Arginine methylation of hnRNPK negatively modulates apoptosis upon DNA damage through local regulation of phosphorylation. Nucleic Acids Res. 42:9908–9924. 2014. View Article : Google Scholar : PubMed/NCBI
38	Feliers D, Lee MJ, Ghosh-Choudhury G, Bomsztyk K and Kasinath BS: Heterogeneous nuclear ribonucleoprotein K contributes to angiotensin II stimulation of vascular endothelial growth factor mRNA translation. Am J Physiol Renal Physiol. 293:F607–F615. 2007. View Article : Google Scholar : PubMed/NCBI
39	Sataranatarajan K, Lee MJ, Mariappan MM and Feliers D: PKCdelta regulates the stimulation of vascular endothelial factor mRNA translation by angiotensin II through hnRNP K. Cell Signal. 20:969–977. 2008. View Article : Google Scholar : PubMed/NCBI