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的一种，是KGF/FGF-7的受体基因。我们明确了涎腺肿瘤在恶变过程中伴随着成纤维细胞生长因子-2的异常表达或过度表达，而且KGFR基因的表达消失，而在涎腺上皮细胞中不表达的FGFR1-IIIc基因通常会随着肿瘤的扩大而增加。我们不仅分析了癌基因和抑癌基因产物的结构或功能的变化，而且还分析了结合蛋白的表达水平和翻译后的修饰变化。利用shRNA表达载体抑制FGFR1-IIIc的表达，从而阻断FGF2-FGFR1-IIIc系统。转导FGFR1-IIIc siRNA的细胞增殖能力下降。利用DNA微阵列和蛋白质组系统分析了野生型KGFR基因在涎腺癌细胞中的表达，发现野生型KGFR基因在涎腺癌细胞中的表达增加了约900个。虽然约有400个基因表达下调。发现与细胞凋亡、肿瘤发生、细胞周期和分子转导相关的基因表达的变化。比较转导KGFR基因的细胞和转导FGFR1siRNA的细胞的基因表达，将以上50个基因重叠后进行蛋白质组互锁FGFR分析。共检测到80多个两次变异的蛋白质点。而FGFR1siRNA可使100多个蛋白质斑点发生改变。

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.