Stromal remodeling in processes of proliferation, invasion and metastasis of oral carcinoma cells : a molecular pathological study

本研究采用间接免疫荧光、生物化学和分子生物学技术，对口腔癌细胞培养过程中细胞外基质分子的生物合成、基质降解酶和细胞外基质（ECM）分子的受体进行了分析。这些口腔癌细胞在体外显示出产生细胞外基质分子。细胞外基质分子的种类和数量因癌细胞而异。尤其是。氨基酸组成和糖基化模式的分子是不同的。这些结果表明，细胞外基质分子的结构差异决定了这些癌细胞在体内的生物学特性。从涎腺腺样囊性癌中建立的ACC 3细胞，其在临床过程中是中等的，以及从具有低转移潜力的舌鳞状细胞癌中建立的ZK-1细胞合成了大量的硫酸乙酰肝素蛋白多糖（HSPG）。相反，从具有高转移潜能的舌鳞状细胞癌建立的MK-1细胞合成稀少量的HSPG及其受体整合素（INT）。ACC-3和MK-1细胞能合成纤维连接蛋白（FN），而ZK-1细胞不能合成。在ACC 3细胞合成的EN中，EDA和EDB区域都包含在它们的mRNA中，而MK-1细胞合成的FN缺乏这些区域，这些区域被交替地剪接出来。在ECM降解酶中，纤溶酶仅由ACC 3细胞合成，而MMP 9在MK-1细胞中合成。在MK-1细胞中，INT α 5、α 2和β 1中的N-连接寡糖比其他细胞中的大5 kDa。这些结果清楚地表明，ECM分子的结构和代谢过程对于细胞粘附到ECM上以及因此口腔癌细胞的临床表型如侵袭性和转移能力是重要的。

We have analyzed the biosynthesis of extracellular matrix molecules, matrix degrading enzymes and receptors for the extracellular matrix (ECM) molecules in oral carcinoma cells in culture by indirect immunofluorescence, biochemical and molecular biological techniques. These oral carcinoma cells were shown to produce extracellular matrix molecules in vitro. The species and amounts of extracellular matrix molecules varied with carcinoma cells. Especially. amino acid compositions and glycosylation patterns of the molecules were distinct. The results suggested that these structural differences in extracellular matrix molecules determined biological characters of those cancer cells in vivo. ACC3 cells established from adenoid cystic carcinoma of the salivary gland, which were moderate in clinical course, as well as ZK-1 cells established from squamous cell carcinoma of tongue with low potential of metastasis synthesized larger amounts of heparan sulfate proteoglycan (HSPG). In contrast, MK-1 cells established from squamous cell carcinoma of tongue with high metastatic potency synthesized scarce amounts of HSPG and its receptor integrin (INT). Fibronectin (FN) was plentifully synthesized by ACC3 and MK-1 cells but not by ZK-I cells. In EN synthesized by ACC3 cells, both EDA and EDB regions were contained in their mRNAs, whereas MK-1 cells synthesized FN lacking those regions which were alternatively spliced-out. Among ECM degradative enzymes, plasmin was synthesized only by ACC3 cells and MMP9 in MK-1 cells. In MK-1 cells, N-linked oligosaccharides in INT alpha5, alpha2 and beta1 were 5 kDa larger than those of other cells. These results clearly indicated that the structures and metabolic processes of ECM molecules were important for cell adhesion onto the ECM and hence clinical phenotypes such as invasiveness and metastatic abilities of oral carcinoma cells.