Umbreit Claudia, Aderhold Christoph, Faber Anne, Sauter Alexander, Hofheinz Ralf-Dieter, Stern-Straeter Jens, Hoermann Karl, Schultz Johannes David
Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68167 Mannheim, Germany.
Department of Hematology and Oncology, University Hospital Mannheim, D-68167 Mannheim, Germany.
Oncol Rep. 2014 Aug;32(2):668-76. doi: 10.3892/or.2014.3225. Epub 2014 May 30.
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.
头颈部鳞状细胞癌（HNSCC）是全球第六大常见癌症类型。HNSCC的生长和侵袭受到细胞外基质（ECM）的强烈影响，而细胞外基质会被基质金属蛋白酶（MMPs）修饰。MMP家族在癌症研究中仍然具有相关性，因为即使在癌症早期，它也能促进恶性转化、细胞增殖和血管生成调节。MMP - 14（MT1 - MMP）对生物活性分子的蛋白水解加工会导致结缔组织严重异常、血管生成缺陷和过早死亡。MMP - 2（明胶酶A）和MMP - 14也在基底膜降解和细胞癌侵袭中发挥作用。伊马替尼可阻断PTK受体c - kit并抑制其PTK活性。本研究的目的是调查基质金属蛋白酶 - 14（MMP - 14）和基质金属蛋白酶 - 2（MMP - 2）在人乳头瘤病毒（HPV）阴性和p16阳性鳞状细胞癌中的表达模式，并评估肿瘤细胞对化疗药物伊马替尼和5 - 氟尿嘧啶（5 - FU）的化疗敏感性。我们用伊马替尼（18和30 μmol/ml）和5 - FU（1和5 μmol/ml）孵育鳞状细胞癌细胞系，并在48、72、120、192和240小时后通过免疫组织化学和酶联免疫吸附测定（ELISA）检测MMP - 14和MMP - 2。我们在所有孵育的肿瘤细胞系中检测到MMP - 2和MMP - 14的表达。特别是使用伊马替尼后，我们发现p16阳性和p16阴性鳞状细胞癌肿瘤细胞系中MMP - 2和MMP - 14表达水平有可靠的下降趋势，此外还有诱导凋亡的作用。我们发现伊马替尼诱导的MMP - 2和MMP - 14抑制具有统计学意义，这取决于孵育时间和细胞系。在用伊马替尼长时间处理后，我们尤其在p16阴性的HNSCC14C细胞中检测到MMP - 2和MMP - 14的显著抑制。伊马替尼和5 - FU的剂量递增对MMP - 2或MMP - 14的表达没有统计学意义上的显著影响。p16阳性鳞状细胞癌细胞表现出更高的总蛋白表达。我们在所有孵育的鳞状细胞癌细胞系中检测到MMP - 2和MMP - 14的显著抑制，部分是在伊马替尼处理后。在用伊马替尼孵育后，我们在CERV196细胞中检测到对MMP - 2的更高抑制。在用伊马替尼处理后，我们发现p16阳性肿瘤细胞在体外的化疗敏感性有可靠的增加趋势。需要进一步的研究和临床试验来在HPV相关的HNSCC中进一步研究这些发现。
