Koybasi Serap, Senkal Can E, Sundararaj Kamala, Spassieva Stefka, Bielawski Jacek, Osta Walid, Day Terry A, Jiang James C, Jazwinski S Michal, Hannun Yusuf A, Obeid Lina M, Ogretmen Besim
Department of Biochemistry, Ralph H. Johnson Veterans Affairs Medical Center, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
J Biol Chem. 2004 Oct 22;279(43):44311-9. doi: 10.1074/jbc.M406920200. Epub 2004 Aug 17.
In this study, endogenous long chain ceramides were measured in 32 human head and neck squamous cell carcinoma (HNSCC) and 10 nonsquamous head and neck carcinoma tumor tissues, as compared with adjacent noncancerous tissues, by liquid chromatography/mass spectroscopy. Interestingly, only one specific ceramide, C(18:0)-ceramide, was selectively down-regulated in the majority of HNSCC tumor tissues. On the other hand, in nonsquamous tumor tissues, this selectivity for C18-ceramide was not detected. These data suggested the hypotheses that decreased levels of C18-ceramide might impart a growth advantage to HNSCC cells and that increased generation of C18-ceramide may be involved in the inhibition of growth. These roles were examined by reconstitution of C18-ceramide at physiologically relevant concentrations in UM-SCC-22A cells (squamous cell carcinoma of hypopharynx) via overexpression of mammalian upstream regulator of growth and differentiation factor 1 (mUOG1), a mouse homologue of longevity assurance gene 1 (mLAG1), which has been shown to specifically induce the generation of C18-ceramide. Liquid chromatography/mass spectroscopy analysis showed that overexpression of the mLAG1/mUOG1 resulted in increased levels of only C(18:0)-ceramide by approximately 2-fold, i.e. concentrations similar to those of normal head and neck tissues. Importantly, increased generation of C18-ceramide by mLAG1/mUOG1 inhibited cell growth (approximately 70-80%), which mechanistically involved the modulation of telomerase activity and induction of apoptotic cell death by mitochondrial dysfunction. In conclusion, this study demonstrates, for the first time, a biological role for LAG1 and C18-ceramide in the regulation of growth of HNSCC.
在本研究中,通过液相色谱/质谱法对32例人类头颈部鳞状细胞癌(HNSCC)和10例非鳞状头颈部癌肿瘤组织中的内源性长链神经酰胺进行了检测,并与相邻的非癌组织进行了比较。有趣的是,在大多数HNSCC肿瘤组织中,只有一种特定的神经酰胺,即C(18:0)-神经酰胺被选择性下调。另一方面,在非鳞状肿瘤组织中,未检测到对C18-神经酰胺的这种选择性。这些数据提示了以下假设:C18-神经酰胺水平降低可能赋予HNSCC细胞生长优势,而C18-神经酰胺生成增加可能参与生长抑制。通过在UM-SCC-22A细胞(下咽鳞状细胞癌)中过表达生长和分化因子1的哺乳动物上游调节因子(mUOG1,长寿保证基因1(mLAG1)的小鼠同源物,已证明其能特异性诱导C18-神经酰胺的生成),以生理相关浓度重建C18-神经酰胺,对这些作用进行了研究。液相色谱/质谱分析表明,mLAG1/mUOG1的过表达仅导致C(18:0)-神经酰胺水平增加约2倍,即与正常头颈部组织相似的浓度。重要的是,mLAG1/mUOG1增加C18-神经酰胺的生成抑制了细胞生长(约70-80%),其机制涉及端粒酶活性的调节和线粒体功能障碍诱导的凋亡性细胞死亡。总之,本研究首次证明了LAG1和C18-神经酰胺在HNSCC生长调节中的生物学作用。