Proteomic analysis of molecular-targeted therapy against KGFR of salivary gland carcinomas

KGF是成纤维细胞生长因子受体之一，是KGF/FGF-7的受体基因。阐明了在涎腺肿瘤恶变过程中伴随着FGF-2的异常表达或过度表达，而且随着恶性程度的增大，KGFR基因表达消失，而FGF-2的受体基因FGFR 1-IIIc在涎腺上皮中通常不表达，随着恶性程度的增大而表达增加。对转染野生型KGFR基因的唾液腺癌细胞和对照癌细胞的基因表达进行了系统和详尽的分析。不仅分析了癌基因和抑癌基因产物的结构或功能变化，而且分析了介导蛋白的表达水平和翻译后的修饰变化。通过使用shRNA表达载体抑制FGFR 1-IIIc表达来阻断FGF 2-FGFR 1-IIIc系统。转染FGFR 1-IIIc siRNA的细胞增殖能力下降。利用DNA微阵列和蛋白质组学系统分析了FGF-FGFR信号调控诱导的涎腺癌细胞分化和凋亡相关基因和蛋白质组，结果发现野生型KGFR基因转染的涎腺癌细胞中约900个基因表达增加。尽管大约400个基因表达减少。基因表达的变化与细胞凋亡、癌变、细胞周期和分子转导有关。比较转染KGFR基因的细胞和转染FGFR 1 siRNA的细胞的基因表达，发现有50多个基因存在重叠。检测到80多个两次变异的蛋白质点。相反，转染FGFR 1 siRNA后，超过100个蛋白点发生了变化。

KGF, one of fibroblast growth factor receptor, is a receptor gene of KGF/FGF-7. We clarified salivary gland tumors accompany the abnormal expression or over-expression of FGF-2 in the process of malignant alteration, furthermore, KGFR gene expression disappeared and FGFR1-IIIc gene, receptor gene of FGF-2 without expression in salivary gland epithelium usually, increase as the enlarging malignancy.In this study, gene expressions were analyzed systematically and exhaustively for salivary gland carcinoma cells transfected wild-type KGFR, gene and control carcinoma cells. And we analyzed not only construction or change in function of oncogene and tumor suppressor gene products but also expression level of interfaced proteins and modified change after translation. FGF2-FGFR1-IIIc system was blocked through inhibition of FGFR1-IIIc expression using shRNA expression vector. The proliferation potency of cells transferred FGFR1-IIIc siRNA decreased. The gene and protein groups by related cell differentiation and apoptosis inducted with manipulation of FGF-FGFR signal were analyzes by DNA micro array and proteome system.In the results, about 900 gene expressions of salivary gland carcinoma transferred wild type KGFR gene were increased. Although about 400 gene expressions were decreased. Variation of gene expressions related apoptosis, carcinogenesis, cell cycle and molecular transducer were recognized. Compared gene expression of cells transfected KGFR gene with cells transfferd FGFR1 siRNA, above 50 genes were overlapped Next proteomic analysis of protein group interlocking FGFR was performed. Above 80 protein spots varied twice were detected. In contrast, above 100 protein spots were changed by transfection of FGFR1 siRNA.