title: Imatinib-associated matrix metalloproteinase suppression in p16-positive squamous cell carcinoma compared to HPV-negative HNSCC cells in vitro.
abstract: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common type of cancer worldwide. The growth and invasion of HNSCC are strongly influenced by the extracellular matrix (ECM), which is modified by matrix metalloproteinases (MMPs). The MMP family is still relevant to cancer research, as it promotes malignant transformation, cell proliferation and modulation of angiogenesis even in the early stages of cancer. The proteolytic processing of bioactive molecules by MMP-14 (MT1-MMP) causes severe abnormalities in connective tissue, defective angiogenesis and premature death. MMP-2 (gelatinase A) and MMP-14 also play a role in degradation of basement membrane and cell carcinoma invasion. Imatinib blocks the PTK receptor c-kit and forestalls its PTK activity. The aim of the present study was to investigate the expression pattern of MMP-14 and MMP-2 in human papilloma virus (HPV)-negative and p16-positive SCC and to evaluate the chemosensitivity of the tumour cells to the chemotherapeutic agents, imatinib and 5-fluorouracil (5-FU). We incubated the SCC cell lines with imatinib (18 and 30 µmol/ml) and 5-FU (1 and 5 µmol/ml) and detected MMP-14 and MMP-2 by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) after 48, 72, 120, 192 and 240 h. We detected expression of MMP-2 and MMP-14 in all incubated tumour cell lines. With imatinib in particular, we found a reliable trend towards decreased MMP-2 and MMP-14 expression levels in p16-positive and p16-negative SCC tumour cell lines in addition to an induced apoptotic effect. We found statistically significant imatinib-induced suppression of MMP-2- and MMP-14, dependent on the incubation time and the cell line. We detected a significant suppression of MMP-2 and MMP-14 especially in p16-negative HNSCC14C cells after prolonged treatment time with imatinib. Dose escalation of imatinib and 5-FU had no statistically significant effect on the expression of MMP-2 or MMP-14. The p16-positive SCC cells exhibited higher expression of total protein. We detected a significant suppression of MMP-2 and MMP-14 in all the incubated SCC cell lines, partially after treatment with imatinib. We found higher suppression of MMP-2 in the CERV196 cells after incubation with imatinib. We detected a reliable trend towards increased chemosensitivity of p16-positive tumour cells in vitro after treatment with imatinib. Extended studies and clinical trials are needed to further investigate these findings in HPV-associated HNSCC.
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Umbreit
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Aderhold
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Faber
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Sauter
authors.affiliation: Department of Hematology and Oncology, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Ralf-Dieter
authors.initials: RD
authors.lastname: Hofheinz
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Stern-Straeter
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Hoermann
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
authors.forename: Johannes David
authors.initials: JD
authors.lastname: Schultz
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000494:pharmacology; D001549:Benzamides / Q000008:administration & dosage* / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000821:virology*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005472:Fluorouracil / Q000008:administration & dosage* / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006258:Head and Neck Neoplasms / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D000068877:Imatinib Mesylate; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D030361:Papillomavirus Infections / Q000188:drug therapy; D010879:Piperazines / Q000008:administration & dosage* / Q000494:pharmacology; D011743:Pyrimidines / Q000008:administration & dosage* / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate; D020782:Matrix Metalloproteinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14; D005472:Fluorouracil