Gene/protein expression of CAPN1/2-CAST system members is associated with ERK1/2 kinases activity as well as progression and clinical outcome in human laryngeal cancer
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Recent evidence indicates the involvement of calpains (CAPNs), a family of cysteine proteases, in cancer development and progression, as well as the insufficient response to cancer therapies. The contribution of CAPNs and regulatory calpastatin (CAST) and ERK1/2 kinases to aggressiveness, disease course, and outcome in laryngeal cancer remains elusive. This study was aimed to evaluate the CAPN1/2-CAST-ERK1/2 enzyme system mRNA/protein level and to investigate whether they can promote the dynamic of tumor growth and prognosis. The mRNA expression of marker genes was determined in 106 laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent mucosa (NCLM) controls using quantitative real-time PCR. The level of corresponding proteins was analyzed by Western Blot. SLUG expression, as indicator of pathological advancement was determined using IHC staining. Significant increases of CAPN1/2-CAST-ERK1/2 levels of mRNA/protein were noted in SCLC compared to NCLM (p < 0.05). As a result, a higher level of CAPN1 and ERK1 genes was related to larger tumor size, more aggressive and deeper growth according to TFG scale and SLUG level (p < 0.05). There were also relationships of CAPN1/2 and ERK1 with incidences of local/nodal recurrences (p < 0.05). An inverse association for CAPN1/2, CAST, and ERK1/2 transcripts was determined with regard to overall survival (p < 0.05). In addition, a higher CAPN1 and phospho-ERK1 protein level was related to higher grade and stage (p < 0.05) and was found to promote worse prognosis. This is the first study to show that activity of CAPN1/2- CAST-ERK1/2 axis may be an indicator of tumor phenotype and unfavorable outcome in SCLC.
KeywordsCAPN1 and CAPN2 genes CAST gene ERK1 and ERK2 genes Calpain-1 and calpain-2 proteins ERK1/2 kinases Calpastatin protein SLUG IHC expression Tumor front grading (TFG) Human laryngeal cancer
This work was supported, in part, by the statutory fund of the Department of Cytobiochemistry, University of Łódź, Poland (506/811) and by a grant from the National Science Council, Poland (N403 043 32/2326).
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- 13.Sundaramoorthy P, Sim JJ, Jang YS, Mishra SK, Jeong KY, Mander P, et al. Modulation of intracellular calcium levels by calcium lactate affects colon cancer cell motility through calcium-dependent calpain. PLoS One. 2015;10(1):e0116984. doi: 10.1371/journal.pone.0116984.CrossRefPubMedPubMedCentralGoogle Scholar
- 14.Rose AH, Huang Z, Mafnas C, Hara JH, Hoffmann FW, Hashimoto AS, et al. Calpain-2 inhibitor therapy reduces murine colitis and colitis-associated cancer. Inflamm Bowel Dis. 2015;21(9):2005–15.Google Scholar
- 28.Salehin D, Fromberg I, Haugk C, Dohmen B, Georg T, Bohle RM, et al. Immunhistochemical analysis for expression of calpain 1, calpain 2 and calpastatin in ovarian cancer. Eur J Gynaecol Oncol. 2012;32(6):628–35.Google Scholar
- 30.Niapour M, Farr C, Minden M, Berger SA. Elevated calpain activity in acute myelogenous leukemia correlates with decreased calpastatin expression. Blood Cancer J. 2012;2(1):e51. doi: 10.1038/bcj.2011.50.
- 41.Howlader NA, Noone AM, Krapcho M, Garshell J, Miller D. SEER Cancer Statistics Review, 1975–2011, National Cancer Institute (2014).Google Scholar
- 44.Paterson EL, Kazenwadel J, Bert AG, Khew-Goodall Y, Ruszkiewicz A, Goodall GJ. Down-regulation of the miRNA-200 family at the invasive front of colorectal cancers with degraded basement membrane indicates EMT is involved in cancer progression. Neoplasia. 2013;15:180–91.CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Starska K, Forma E, Jóźwiak P, Bryś M, Lewy-Trenda I, Brzezińska-Błaszczyk E, et al. Gene and protein expression of glucose transporter 1 and glucose transporter 3 in human laryngeal cancer-the relationship with regulatory hypoxia-inducible factor-1α expression, tumor invasiveness, and patient prognosis. Tumour Biol. 2015;36(4):2309–21.CrossRefPubMedGoogle Scholar
- 46.Starska K, Forma E, Nowacka-Zawisza M, Lewy-Trenda I, Ciesielski P, Pietruszewska W, et al. The c.*229C > T gene polymorphism in 3′UTR region of the topoisomerase IIβ binding protein 1 gene and LOH in BRCA1/2 regions and their effect on the risk and progression of human laryngeal carcinoma. Tumour Biol. 2015. doi: 10.1007/s13277-015-4276-3.PubMedCentralGoogle Scholar