Cyto-biological study for growth mechanism of oral squamous cell carcinoma cells in protein-free culture

口腔鳞癌的预后取决于肿瘤细胞的侵袭和转移。为了阐明肿瘤的侵袭和转移机制，分析肿瘤及其周围组织产生的细胞因子和蛋白水解酶的功能是非常重要的。此外，比较SOC细胞和正常口腔角质形成细胞的特性也很重要。癌细胞已在添加胎牛血清的培养液中培养，这与正常口腔角质形成细胞的培养液不同。随着端粒DNA的缩短，正常细胞的特性也会随着细胞分裂而发生变化。因此，有必要研制一种既能支持癌细胞生长又能支持正常口腔角质形成细胞生长的无蛋白培养液，并建立与正常细胞一样具有稳定细胞生物学特性的永生化口腔角质形成细胞。用CONT-…支持口腔黏膜上皮细胞和正常口腔角质形成细胞的生长更多滚动的钙浓度。此外，我们还发现，表达转录因子Snail的SOC细胞缺乏E-钙粘素的表达，并且侵袭性更强。Snail与上皮-间充质转化(ENT)有关。用FLDS分析EGF和HGF对SOC细胞生长的影响，发现这两种生长因子仅对Snail阴性和E-cadherin阳性的SOC细胞有抑制作用。此外，我们还发现，蜗牛诱导的EMT增加了基质金属蛋白酶-2的表达。此外，在对正常口腔上皮细胞、癌前病变细胞和恶性口腔上皮细胞中端粒酶及其相关蛋白的活性和表达进行了精确的研究后，我们成功地将人端粒酶催化成分hTERT和HPV16-E7导入口腔角质形成细胞，建立了永生化的口腔角质形成细胞。这些细胞在添加EGF和牛脑垂体提取物的培养液中具有正常的分化能力。我们的结果使我们能够在相同的无蛋白介质中准确地比较SOC细胞和永生化口腔角质形成细胞。较少

Prognosis of oral squamous cell carcinomas (SOC) depends on tumor cell invasion and metastasis. In order to elucidate the mechanisms of invasion and metastasis, it is important to analyze the functions of cytokines and proteases produced by cancer and the surrounding tissues. Furthermore, it is important to compare characters of SOC cells with normal oral keratinocytes. Cancer cells have been cultured in the media supplemented with fetal bovine serum which are different from the media for normal oral keratinocytes. Also characters of normal cells change by cell divisions with shortening of telomere DNA. Therefore it is necessary to produce protein free medium which support growth of both cancer cells and normal oral keratinocytes and to establish immortalized oral keratinocytes maintaining stable cyto-biological characters as normal cells.In this study, we first produced a new protein free medium, FLDS. This medium supported the growth of SOC cells and normal oral keratinocytes by cont … More rolling calcium concentration. Furthermore, we demonstrated that SOC cells expressing a transcriptional factor, Snail, which relates to the epithelial-mesenchymal transition (ENT), lacked E-cadherin expression, and were more invasive. Analysis of the effects of EGF and HGF using FLDS medium revealed these two growth factors inhibited only Snail negative and E-cadherin positive SOC cells. Also, we found that expression of matrix metalloproteinase-2 was increased by Snail-induced EMT. Furthermore, after the precise studies of the activity and expressions of telomerase and its associated proteins in normal, premalignant and malignant oral epithelial cells, we succeeded to establish immortalized oral keratinocytes by transfect ion with hTERT, a catalytic component of human telomerase and HPVl6-E7 These cells exhibited normal differentiation abilities in the medium supplemented with EGF and bovine pituitary extract. Our results enable us to compare SOC cells and immortalized oral keratinocytes in the same protein-free medium precisely. Less