Do NFATc2 and cJun Cooperate to Activate Transcription in Breast Cancer Cells?

描述（由申请人提供）：乳腺癌是西方世界女性最常见的癌症，也是全球女性癌症相关死亡的主要原因。许多已知与乳腺癌有关的分子因素最终会引起它们的影响 通过引起基因表达的变化。调控基因表达的关键控制点是mRNA转录水平。为了控制乳腺癌肿瘤的不受抑制的生长和侵袭性，必须了解对疾病进展至关重要的基因转录水平的疾病相关扰动。转录激活因子NFATc 2和cJun已分别显示激活增加乳腺癌细胞侵袭力的基因的转录。NFATc 2和cJun是否合作激活乳腺癌细胞中的转录尚不清楚，尽管它们在控制其他生物系统中的转录程序方面的合作是公认的。NFATc 2的细胞内定位受细胞溶质钙水平控制。几种化学 已知致癌物增加乳腺上皮细胞或乳腺癌细胞中的胞质钙。这些化学物质是否导致NFATc 2定位于细胞核尚不清楚。这项初步研究将解决NFATc 2和cJun合作激活基因转录的假设，这些基因促进乳腺癌侵袭性，以应对已知增加细胞溶质钙的环境致癌物。该提案有三个高度集中的具体目标。1)通过使用免疫荧光确定已知可增加细胞溶质钙对NFATc 2核定位的环境致癌物的影响。2)在用致癌物处理乳腺上皮细胞或乳腺癌细胞后，使用ChIP-seq鉴定NFATc 2和cJun共定位的基因。3)确定NFATc 2和cJun是否协同上调基因的表达，这些基因在用致癌物处理后对细胞生长和侵袭性很重要，通过敲低转录激活因子并确定对选定基因的mRNA水平的影响。这项拟议的研究将确定NFATc 2和cJun是否合作激活转录，以响应用化学致癌物处理细胞。尽管这些研究主要集中在乳腺癌上，因为有明确的证据表明NFATc 2和cJun促进了乳腺癌细胞的侵袭性，但这项研究也将影响其他癌症类型的研究，这些癌症类型也被认为是这些因素的作用。这些试点研究的结果将被用作使用基于细胞和生物化学研究的新研究路线的基础，以揭示NFATc 2和cJun激活乳腺癌转录的机制。

DESCRIPTION (provided by applicant): Breast cancer is the most common cancer in women in the Western world, and is the leading cause of cancer related deaths in women worldwide. Many of the molecular factors known to be involved in breast cancer ultimately elicit their effects by causing changes in gene expression. A critical control point for regulating gene expression is at the level of mRNA transcription. In order to control the unchecked growth and invasiveness of breast cancer tumors, an understanding of disease-related perturbations in the transcription levels of genes critical to the progression of the disease must be obtained. The transcriptional activators NFATc2 and cJun have each individually been shown to activate transcription of genes that increase breast cancer cell invasiveness. Whether NFATc2 and cJun cooperate to activate transcription in breast cancer cells is not known, although their cooperation in controlling transcriptional programs in other biological systems is well-established. The intracellular localization of NFATc2 is controlled by levels of cytosolic calcium. Several chemical carcinogens are known to increase cytosolic calcium in mammary epithelial cells or breast cancer cells. Whether these chemicals cause NFATc2 to localize to the nucleus is not known. This pilot study will address the hypothesis that NFATc2 and cJun cooperate to activate transcription of genes that promote breast cancer invasiveness in response to environmental carcinogens known to increase cytosolic calcium. The proposal has three highly focused Specific Aims. 1) Determine the effect of environmental carcinogens known to increase cytosolic calcium on NFATc2 nuclear localization through the use of immunofluorescence. 2) Identify genes at which NFATc2 and cJun co-localize using ChIP-seq after treatment of mammary epithelial cells or breast cancer cells with a carcinogen. 3) Determine whether NFATc2 and cJun cooperate to upregulate expression of genes important to cell growth and invasiveness upon treatment with a carcinogen by knocking down the transcriptional activators and determining the effect on mRNA levels of select genes. The proposed research will determine whether NFATc2 and cJun cooperate to activate transcription in response to treating cells with a chemical carcinogen. Although the studies are focused on breast cancer because of the clear evidence that NFATc2 and cJun promote invasiveness of breast cancer cells, the research will also influence studies of other cancer types in which these factors are also thought to work. The results of these pilot studies will be used as a foundation for new lines of research using cell-based and biochemical studies to unravel the mechanisms by which NFATc2 and cJun activate transcription in breast cancer.