Comparative analyses of MYBL1 knockdown in non-tumor and triple negative breast cancer cells

非肿瘤性乳腺癌与三阴性乳腺癌MYBL1基因敲除的对比分析 单元格 项目总结 MYBL1基因属于MYB基因家族，包括c-MYB和MYBL2。基因 原癌基因作为强大的转录激活剂参与了细胞增殖， 分化和细胞周期信号传递过程，所有这些都是肿瘤进展的关键事件。研究 表明截短的c-myb是一种癌基因，因此，该基因被研究为治疗的靶点。 乳腺癌。对MYBL1和MYBL2知之甚少。两者在细胞周期中都扮演着重要的角色 并在不同的癌症中过度表达，包括乳腺癌。发布的结果来自 Player实验室显示MYBL1在三阴性乳腺癌亚群中过表达 (TNBC)。在一项单独的综合监督网络分析中，MYBL1对管腔性乳腺癌 研究表明，过度表达与疾病预后不良相关。基于这些研究，校长 调查者(PI)实验室选择研究MYBL1基因，对该基因的特征感兴趣 在非肿瘤TN和TNBC中进行比较，并确定其在肿瘤中表达的影响基因。这个 实验方法是敲除样本中的MYBL1基因，进行微阵列分析 确定受这些过程差异影响的基因，然后进行进一步的分析以进行验证 在这个过程中确定了候选基因。作为概念的证明，MYBL1基因被下调 在TNBC细胞系中。受MYBL1表达降低影响的候选基因包括MYBL2和 细胞周期信号基因的丰富，其中一些是新的。这一观察结果与 显示MYBL1和MYBL2共表达并证实MYBL1有能力 调控MYBL2的表达。目前研究中概述的实验应该会扩大我们的 了解三重阴性乳房样本中的信号事件。对于目标1，调查人员将 继续在TNBC中对MYBL1进行敲除研究，并包括一个额外的TNBC和一个非肿瘤TN 样本作为比较。这些数据将使PI能够通过以下方式识别不同的监管模式 MYBL1在非肿瘤和三重阴性癌症中的比较努力识别 可能与肿瘤的进展有关。因为TNBC是异质的，所以这项研究仅限于 B亚型、MYBL1表达样本。可靠的标志性基因将被识别，并进行路径分析 已执行。假设MYBL1在不同的组织中表达会导致不同的 信令机制。选择差异表达的基因将使用组织微阵列进行检查 并与MYBL1基因表达模式进行比较。为 目的2、不同击倒样品的增殖能力和侵袭潜能 比较一下。目的3对照未处理、shRNA干扰对照和 每个细胞系的MYBL1 shRNA敲除制剂将通过软琼脂分析进行检测。

Comparative analyses of MYBL1 knockdown in non-tumor and triple negative breast cancer cells PROJECT SUMMARY The MYBL1 gene belongs to the MYB family of genes, which includes c-MYB and MYBL2. The genes are proto-oncogenes that function as strong transcriptional activators involved in proliferation, differentiation and cell cycle signaling processes, all which are key events in tumor progression. Studies showed that truncated c-MYB is an oncogene and as such, the gene was studied as a therapeutic target for breast cancers. Less is known about MYBL1 and MYBL2. Both play substantial roles in cell cycle progression and are overexpressed in different cancers, including breast cancers. Published results from the Player laboratory showed MYBL1 overexpressed in a subpopulation of triple negative breast cancers (TNBC). In a separate comprehensive supervised network analyses of luminal breast cancers, MYBL1 overexpression was shown to correlate with poor disease prognosis. Based on these studies the principal investigator‘s (PI) laboratory chose to study the MYBL1 gene with an interest in characterizing the gene in non-tumor TN compared to TNBC and identifying genes affected by its expression in the cancers. The experimental approach is to knockdown the MYBL1 gene in the samples, perform microarray analyses to identify genes differentially affected by the processes, followed by further analyses to authenticate candidate genes identified during the process. As a proof of concept, MYBL1 gene was down-regulated in a TNBC cell line. The candidate genes affected by decreasing MYBL1 expression include MYBL2 and an enrichment in cell cycle signaling genes some of which are novel. This observation is consistent with those which show co-expression of MYBL1 and MYBL2 and substantiate the ability of MYBL1 to regulate expression of MYBL2. The experiments outlined in the current study should broaden our understanding of signaling events in triple negative breast samples. For Aim 1, the investigators will continue knockdown studies of MYBL1 in TNBC and include an additional TNBC and a non-tumor TN sample as comparisons. These data will allow the PI to identify differential patterns of regulation by MYBL1 in non-tumor compared to triple negative cancers in efforts to identify genes and processes that might be associated with tumor progression. Because TNBC are heterogeneous, this study is confined to subtype B, MYBL1 expressing samples. Reliable signature genes will be identified, and pathway analyses performed. The hypothesis is that expression of MYBL1 in the different tissues result in different signaling mechanisms. Select differentially expressed genes will be examined using tissue microarrays with defined receptor status and survival data and compared to the MYBL1 gene expression pattern. For Aim 2, the proliferative capacity and invasive potential of the various knockdown samples will be compared. For Aim 3 the tumorigenic properties of control untreated, shRNA scrambled control and MYBL1 shRNA knockdown preparations for each cell line will be examined via Soft Agar analyses.