Scatter factor induced carcinoma cell migration

散射因子诱导癌细胞迁移

• 批准号: 6681190
• 负责人: J. Suzanne Lindsey
• 金额: $ 11.1万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6681190
• 关键词: antisense nucleic acid; biological signal transduction; blocking antibody; carcinoma; cell adhesion molecules; cell migration; chemotaxis; complementary DNA; enzyme linked immunosorbent assay; extracellular matrix; focal adhesion kinase; gene expression; hepatocyte growth factor; immunocytochemistry; integrins; metastasis; neoplasm /cancer genetics; protein sequence; protooncogene

DESCRIPTION (provided by applicant): Interactions between growth factors secreted by the stroma cells, extracellular matrix, and their receptors on carcinoma cells resulting in signal transduction are key to understanding the mechanisms underlying carcinoma cell migration and invasion. Such understanding requires identification of the genes induced by these interactions and the convergence of signaling pathways. Scatter factor, also known as Hepatocyte growth factor, (SF) and its tyrosine kinase protooncogene, Met, are upregulated in most metastatic cancers and is associated with a poor prognosis. SF causes migration of cancer cells, a process that requires de novo gene transcription. However, the early genes expressed by this new transcription are not known. Our long range goal is to determine carcinoma cell specific targets to inhibit metastasis. We have isolated a novel cDNA we call Mig-7 that is induced by SF in the human endometrial carcinoma cell lines, RL-95 [and HECIA, as well as the pancreatic carcinoma cell line FG] before migration occurs. Homology searches show that Mig-7 cDNA sequence possesses 99% homology with two human ESTs. However, no full length cDNA, gene or protein homologies were found. Expression of this gene is lacking in normal tissue as determined by reverse transcription and polymerase chain reaction. In contrast, it is detected in various metastatic human tumors such as ovary, colon, endometrial and squamous cell. Furthermore, the Mig-7 induction by SF is also regulated by ?v?5 integrin ligation. The focus of this proposal is to understand the role of Mig-7 in expression in carcinoma cell migration. Our hypothesis is that Mig-7 expression plays a role in migration of cancer cells. To test this hypothesis and accomplish the objective of this application we will pursue the following two specific aims: Aim 1 is to determine if the degree of cell migration is dependent upon the level of Mig-7 expression in carcinoma cells to test our hypothesis that Mig-7 expression leads to migration of carcinoma cells. [We base our hypothesis on our preliminary results showing that we can inhibit carcinoma cell migration by 83.50% +/- 2.77% (p<0.05) in vitro using antisense oligonucleotides specific to Mig-7 as compared to irrelevant oligonucleotide.] We will overexpress Mig-7 in carcinoma cell lines and measure migration and invasion capabilities as compared to controls using migration [and invasion] assays. Aim 2 is to determine the signaling mechanism underlying the cross talk between the Met and alphavbeta5 integrln signal transduction pathways because our studies indicate that both Met and alphavbeta5 integrin signaling pathways are required to induce Mig-7 expression. [Met activation has been shown to activate focal adhesion kinase (FAK). Further, tyrosine kinase receptor induction of FG carcinoma cell migration requires alphavbeta5 integrin binding. However, gene expression resulting from this cross-talk signaling has not been determined.] Mig-7 is a novel gene expressed in this signaling system. We expect that these studies will lead to a significant advance in the knowledge of how carcinoma cells migrate and could lead to a target for new anti-metastatic therapy.

描述（由申请人提供）：基质细胞分泌的生长因子，细胞外基质及其受体在癌细胞上导致信号转导的受体是了解癌细胞迁移和侵袭的基础机制的关键。这种理解需要鉴定由这些相互作用引起的基因以及信号通路的收敛性。散射因子，也称为肝细胞生长因子（SF）及其酪氨酸激酶原生物元，在大多数转移性癌症中被上调，并且预后不良。 SF引起癌细胞的迁移，这一过程需要从头基因转录。但是，该新转录表达的早期基因尚不清楚。我们的远距离目标是确定癌细胞特异性靶标抑制转移。我们分离了一种新型的cDNA，我们称为MIG-7，该cDNA在人子宫内膜癌细胞系中由SF诱导，RL-95 [和HECIA以及HECIA，以及胰腺癌细胞系FG]。同源性搜索表明，MIG-7 cDNA序列与两个人类EST具有99％同源性。但是，未发现全长cDNA，基因或蛋白质同源性。正常组织中缺乏该基因的表达，这是由逆转录和聚合酶链反应确定的。相反，在各种转移性人类肿瘤（例如卵巢，结肠，子宫内膜和鳞状细胞）中检测到它。此外，SF的MIG-7诱导也受“ V？5整联蛋白连接”的调节。该建议的重点是了解MIG-7在表达在癌细胞迁移中的作用。我们的假设是MIG-7表达在癌细胞的迁移中起作用。为了检验这一假设并实现该应用的目标，我们将追求以下两个特定目的：目标1是确定细胞迁移程度是否取决于癌细胞中MIG-7表达水平，以检验我们的假设，即MIG-7表达导致癌细胞的迁移。 [我们的假设基于初步结果，表明我们可以在体外抑制83.50％+/- 2.77％（p <0.05），使用反义寡核苷酸与MIG-7特定的反义寡核苷酸在体外抑制癌细胞的迁移，而与非核苷酸的相比，我们将使用co-7的偏态迁移和测量的迁移。 [和入侵]测定法。 AIM 2是确定MET和Alphavbeta5 Integrln信号转导途径之间交叉讲座的信号传导机制，因为我们的研究表明Met和Alphavbeta5整合素信号通路都需要诱导MIG-7表达。 [MET激活已被证明可以激活局灶性粘附激酶（FAK）。此外，FG癌细胞迁移的酪氨酸激酶受体诱导需要alphavbeta5整合素结合。然而，尚未确定由这种串扰信号引起的基因表达。] MIG-7是在该信号系统中表达的一种新基因。我们预计这些研究将导致对癌细胞如何迁移的知识有重大进步，并可能导致新的抗转移疗法的靶标。