EXPRESSION AND FUNCTIONAL-ACTIVITY OF FIBROBLAST GROWTH-FACTORS AND THEIR RECEPTORS IN HUMAN PANCREATIC-CANCER
EXPRESSION AND FUNCTIONAL-ACTIVITY OF FIBROBLAST GROWTH-FACTORS AND THEIR RECEPTORS IN HUMAN PANCREATIC-CANCER
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DOI：
10.1002/ijc.2910590515
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发表时间：
1994-12-01
影响因子：
6.4
通讯作者：
LEMOINE, NR
中科院分区：
文献类型：
--
作者：
LEUNG, HY;GULLICK, WJ;LEMOINE, NR
我们分析了肝素结合成纤维细胞生长因子（FGF）家族的前7个成员及其4个高亲和力受体（FGFR）在人胰腺癌细胞系中的表达，包括mRNA和蛋白水平。在表达FGFR的细胞系中，观察到2种典型模式：（i）FGFR-1、FGFR-3或FGFR-4沿着至少一种FGF的表达;（ii）在不存在FGF表达的情况下FGFR-3和FGFR-4的共表达。使用RT-PCR，在细胞系PT 45中检测到代表FGFR-1的细胞外和细胞内结构域的多种同种型的转录物。预测FGFR-1的新型胞外域变体以潜在分泌形式编码第一免疫球蛋白环。在放射性标记的配体交联实验中，通过用特异性抗体进行免疫沉淀来确认FGFR-1的剪接变体的蛋白表达。在PANC-1细胞系中检测到1型羧基端和α亚型胞外结构域，而在PT 45细胞系中检测到1型羧基端和γ亚型胞外结构域。还使用3种不同的抗FGF-2抗体通过免疫沉淀检测PT 45中FGF-2的表达。除了18-kDa产物外，还表达了更高分子量的同种型，即22-和23-kDa同种型。在锚定非依赖性生长的测定中，外源性FGF-2分别刺激细胞系MIA PACA-2和PANC-1的集落形成最多增加15倍和10倍。相同细胞系的单层培养物的处理没有促进生长。然而，针对FGF-2的特异性中和抗体使MIA PACA-2细胞的细胞增殖降低了50%。(C)1994 Wiley-Liss，Inc.
We have analysed expression of the first 7 members of the family of heparin-binding fibroblast growth factor (FGFs) and their 4 high-affinity receptors (FGFRs) in human pancreatic carcinoma cell lines, both at the mRNA and protein levels. In cell lines expressing FGFRs, 2 typical patterns were observed: (i) expression of FGFR-1, -3 or -4 along with the expression of at least one FGF; (ii) co-expression of FGFR-3 and FGFR-4 in the absence of FGF expression. Using RT-PCR, transcripts representing multiple isoforms of both extracellular and intracellular domains of FGFR-1 were detected in the cell line PT45. A novel extracellular domain variant of FGFR-1 was predicted to encode the first immunoglobulin loop in a potentially secreted form. Protein expression of the splice variants of FGFR-1 was confirmed by immunoprecipitation with specific antibodies in radiolabelled ligand cross-linking experiments. The type carboxyl end and the alpha subtype extracellular domain were detected in the PANC-1 cell line, while the type 1 carboxyl terminus and the gamma subtype extracellular domain were expressed in the PT45 cell line. Expression of FGF-2 in PT45 was also detected by immunoprecipitation using 3 different anti-FGF-2 antibodies. Apart from the 18-kDa product, higher molecular weight isoforms, namely 22- and 23-kDa isoforms, were expressed. In an assay of anchorage-independent growth, exogenous FGF-2 stimulated a maximum 15-fold and 10-fold increase in colony formation by the cell lines MIA PACA-2 and PANC-1 respectively. Treatment of monolayer cultures of the same cell lines did not promote growth. However, a specific neutralising antibody against FGF-2 reduced cell proliferation of MIA PACA-2 cells by 50%. (C) 1994 Wiley-Liss, Inc.