title: 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.
abstract: 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.
authors.affiliation: I Department of Otolaryngology and Laryngological Oncology, Medical University of Łódź, Kopcińskiego 22, 90-153, Lodz, Poland. katarzyna.starska@umed.lodz.pl.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Starska
authors.affiliation: Department of Cytobiochemistry, University of Łódź, Pomorska 142/143, 90-236, Lodz, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Forma
authors.affiliation: Department of Cytobiochemistry, University of Łódź, Pomorska 142/143, 90-236, Lodz, Poland.
authors.forename: Paweł
authors.initials: P
authors.lastname: Jóźwiak
authors.affiliation: Department of Pathology, Medical University of Łódź, Pomorska 251, 92-213, Lodz, Poland.
authors.forename: Iwona
authors.initials: I
authors.lastname: Lewy-Trenda
authors.affiliation: Department of Pathology, Medical University of Łódź, Pomorska 251, 92-213, Lodz, Poland.
authors.forename: Marian
authors.initials: M
authors.lastname: Danilewicz
authors.affiliation: Department of Nephropathology, Medical University of Łódź, Pomorska 251, 92-213, Lodz, Poland.
authors.forename: Olga
authors.initials: O
authors.lastname: Stasikowska-Kanicka
authors.affiliation: Department of Otolaryngology, Żeromski Specialist Hospital, os. Na Skarpie 66, 31-913, Krakow, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Skóra
authors.affiliation: I Department of Otolaryngology and Laryngological Oncology, Student Scientific Circle of Laryngological, Immunobiology, Medical University of Łódź, Kopcińskiego 22, 90-153, Lodz, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kolary
authors.affiliation: I Department of Otolaryngology and Laryngological Oncology, Student Scientific Circle of Laryngological, Immunobiology, Medical University of Łódź, Kopcińskiego 22, 90-153, Lodz, Poland.
authors.forename: Jakub
authors.initials: J
authors.lastname: Miazga
authors.affiliation: Department of Cytobiochemistry, University of Łódź, Pomorska 142/143, 90-236, Lodz, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Krześlak
authors.affiliation: Department of Cytobiochemistry, University of Łódź, Pomorska 142/143, 90-236, Lodz, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Bryś
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D018450:Disease Progression; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D007820:Laryngeal Mucosa / Q000378:metabolism / Q000473:pathology; D007822:Laryngeal Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000401:mortality / Q000473:pathology; D008297:Male; D008875:Middle Aged; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D060787:Neoplasm Grading; D009364:Neoplasm Recurrence, Local; D010641:Phenotype; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012307:Risk Factors; D047368:Tumor Burden
chemical_list: D002135:Calcium-Binding Proteins; D012333:RNA, Messenger; C033668:calpastatin; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002154:Calpain; C484806:CAPN1 protein, human; C569870:CAPN2 protein, human
keywords: CAPN1 and CAPN2 genes; CAST gene; Calpain-1 and calpain-2 proteins; Calpastatin protein; ERK1 and ERK2 genes; ERK1/2 kinases; Human laryngeal cancer; SLUG IHC expression; Tumor front grading (TFG)