The Src Kinase, cell communication & growth control

Src 激酶，细胞通讯

• 批准号: 6780978
• 负责人: GARY S GOLDBERG
• 金额: $ 21.45万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6780978
• 关键词: biological signal transduction; cell growth regulation; cell transformation; electrical conductance; enzyme activity; fluorescent dye /probe; gap junctions; gene expression; immunofluorescence technique; intercellular connection; membrane channels; microarray technology; mitogen activated protein kinase; mixed tissue /cell culture; neoplastic cell; protein kinase; transfection; voltage /patch clamp

DESCRIPTION (provided by applicant): Tumor cell growth can be controlled by surrounding normal cells. This process is called "heterologous growth control". The goal of this project is to elucidate mechanisms underlying this process. We will determine how nontransformed cells affect the Src signaling cascade in neighboring transformed cells. The need for heterologous gap junctional communication in heterologous growth control will be evaluated in Specific Aim 1. We hypothesize that nontransformed cells do not need to form active gap junction channels with Src transformed cells to normalize their growth. We will utilize connexin knockout cells, chemical blockers or antisense nucleic acids to suppress gap junctional communication. Intercellular communication will be examined by the transfer of fluorescent dyes, endogenous metabolites, and electrical conductance. If heterologous growth control occurs in the absence of gap junctional communication, then, as we hypothesize, gap junctional communication is not required for heterologous growth control. Effects of nontransformed cells on the Src kinase activity, and the activity of kinases acting downstream of Src, in Src transformed cells will be evaluated in Specific Aims 2 and 3, respectively. We hypothesize that nontransformed cells quell Src kinase activity, and affect the activity of downstream kinases, in neighboring transformed cells. We will use novel techniques that we have developed to examine kinase activity in transformed cells that are morphologically reversed to a normal phenotype by neighboring nontransformed cells. Our hypothesis will be proven if the Src signaling in transformed cells cascade is blocked by communication with nontransformed cells. Effects of nontransformed cells on the global expression pattern of genes in neighboring transformed cells will be examined in Specific Aim 4. We hypothesize that nontransformed cells alter the expression of specific genes in neighboring transformed cells. We will utilize DNA chips to compare the gene profiles of transformed cells cocultured with nontransformed cells to transformed cells and nontransformed cells grown alone. The hypothesis will be proven if the expression levels of some genes, but not others, in transformed cells are affected by contact with nontransformed cells during heterologous growth control.

描述(申请人提供)：肿瘤细胞生长可由周围正常细胞控制。这个过程被称为“异源生长控制”。这个项目的目标是阐明这一过程背后的机制。我们将确定未转化细胞如何影响邻近转化细胞中的Src信号级联。在异源生长控制中对异源缝隙连接通信的需求将在特定的目标1中进行评估。我们假设未转化的细胞不需要与Src转化的细胞形成活跃的缝隙连接通道来使其正常生长。我们将利用连接蛋白敲除细胞、化学阻滞剂或反义核酸来抑制缝隙连接通讯。细胞间的通讯将通过荧光染料、内源性代谢物和电导的转移来检测。如果异源生长控制是在没有缝隙连接通讯的情况下发生的，那么，正如我们假设的那样，异源生长控制不需要缝隙连接通讯。在特定的目标2和3中，将分别评估未转化细胞对Src激酶活性的影响，以及在Src转化细胞中作用于Src下游的激酶的活性。我们假设，在邻近的转化细胞中，未转化的细胞抑制了Src激酶的活性，并影响了下游的激酶的活性。我们将使用我们开发的新技术来检测转化细胞中的激酶活性，这些转化细胞在形态上被邻近的未转化细胞逆转为正常表型。如果转化细胞中的Src信号级联被与未转化细胞的通讯阻断，我们的假设将得到证实。未转化细胞对邻近转化细胞中基因整体表达模式的影响将在特定目标4中进行考察。我们假设未转化细胞改变了邻近转化细胞中特定基因的表达。我们将利用DNA芯片将与未转化细胞共培养的转化细胞与转化细胞和单独生长的未转化细胞的基因图谱进行比较。如果在异源生长控制过程中，转化细胞中某些基因的表达水平受到与未转化细胞接触的影响，这一假说将得到证实。