Effects of Cytokines on the Proliferation Mechanism of Oral Cancer and Oral Epithelial Cells

我们已经成功地培养了许多人类癌细胞，包括口腔鳞状细胞癌（SCC）细胞在无蛋白质的合成培养基，PF 86 -1。然而，有一些人类癌细胞不能在无蛋白质条件下增殖，考虑到可能缺乏生长的必需因子。在这项研究中，它是调查是否细胞因子可以作为口腔鳞状细胞癌细胞在无蛋白条件下培养的生长刺激。因此，IL-1a通过自分泌和旁分泌机制对口腔鳞状细胞癌细胞起生长刺激作用。最近，我们开发了另一种新的无蛋白合成培养基PFM-7，用于培养正常人口腔角质形成细胞。在PFM-7中生长的正常口腔角质形成细胞与在含有生长因子的常规培养基中培养的细胞相比，在形态、对添加的生长因子的反应以及生长因子及其受体的基因表达方面没有变化的情况下，在培养中生长表现出几乎相等的效力。利用这些无蛋白条件，对同一患者的SCC细胞系和正常口腔角质形成细胞的基因表达进行了比较研究，构建了正常和恶性细胞的差减cDNA文库，分离出癌细胞中与同一患者的正常口腔角质形成细胞相比增加或减少的特定基因。其中一个在癌细胞中被抑制的基因与S100蛋白具有同源性，并且在RNA印迹杂交分析中在几种SCC细胞系中被抑制。目前对该基因的功能进行了分析。

We have succeeded in culturing many human cancer cells including oral squamous cell carcinoma (SCC) cells in a protein-free synthetic medium, PF86-1. However, there are some human cancer cells that can't proliferate under protein-free conditions, It is contemplated that the essential factors for the growth may be lacking. In this study, it was investigated whether cytokines could act as a growth stimulator for oral squamous cell carcinoma cells cultured under protein-free conditions. As a result, IL-1a acts as a growth stimulator for oral squamous cell carcinoma cells by autocrine and paracrine mechanisms.Recently, another novel protein-free synthetic medium, PFM-7 has been developed by us for the culture of normal human oral keratinocytes. Normal oral keratinocytes growing in PFM-7 exhibited nearly equal potency to grow in culture without changes in morphology, response to the added growth factors and gene expression of growth factors and their receptors when compared with the cells cultured in the conventional medium containing growth factors. Using these protein-free conditions, a comparative study of gene expression of SCC cell lines and normal oral keratinocytes derived from the same patient was performed.Subtractive cDNA libraries of normal and malignant cells were constructed and specific genes which are increased or decreased in cancer cells compared to the normal oral keratinocytes of the same patient were isolated. One of the genes which are suppressed in cancer cells had a homology of S100 protein and is decreased in several SCC cell lines in RNA blot hybridization analysis. Functions of the gene are analized now.