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

非肿瘤细胞和三阴性乳腺癌细胞中 MYBL1 敲低的比较分析

• 批准号: 10415624
• 负责人: Audrey Player
• 金额: $ 13.92万
• 依托单位: TEXAS SOUTHERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-06 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10415624
• 关键词: Affect; Agar; Biological; Biological Assay; Biological Response Modifier Therapy; Breast; Breast Cancer Cell; Breast Cancer cell line; Burkitt Lymphoma; Cancer Patient; Candidate Disease Gene; Cell Cycle; Cell Cycle Progression; Cell Line; Cells; Characteristics; Colon; Contact Inhibition; Data; Data Analyses; Dependence; Down-Regulation; Event; Gene Expression Profile; Gene Family; Genes; Immunohistochemistry; In Vitro; Knowledge; Laboratories; MCF10A cells; MYB gene; MYBL1 gene; MYBL2 gene; Malignant Neoplasms; Mammary Neoplasms; Microarray Analysis; Oncogenes; Pathway Analysis; Patients; Pattern; Play; Preparation; Principal Investigator; Procedures; Process; Property; Proto-Oncogenes; Publishing; Regulation; Research Personnel; Role; Sampling; Signal Transduction; Signaling Protein; Statistical Data Interpretation; Supervision; Testing; Tissue Microarray; Tissues; Transcription Coactivator; Uterine Fibroids; Validation; aggressive breast cancer; base; cancer subtypes; cancer type; comparative; design; differential expression; disease prognosis; experimental study; genetic signature; interest; knock-down; malignant breast neoplasm; neoplastic cell; novel; overexpression; receptor; screening; small hairpin RNA; therapeutic target; transcription factor; transcriptome; triple-negative invasive breast carcinoma; tumor progression; tumorigenic

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.

非肿瘤和三阴性乳腺癌中 MYBL1 敲低的比较分析 细胞 项目概要 MYBL1 基因属于 MYB 基因家族，该家族包括 c-MYB 和 MYBL2。基因 是原癌基因，作为参与增殖的强转录激活剂， 分化和细胞周期信号传导过程，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，因此该基因被研究作为治疗靶点 乳腺癌。人们对 MYBL1 和 MYBL2 知之甚少。两者都在细胞周期中发挥重要作用 进展并在不同的癌症中过度表达，包括乳腺癌。发布结果来自 Player 实验室显示 MYBL1 在三阴性乳腺癌亚群中过度表达 （TNBC）。在一项单独的管腔乳腺癌综合监督网络分析中，MYBL1 过度表达被证明与不良的疾病预后相关。基于这些研究主要 研究人员 (PI) 实验室选择研究 MYBL1 基因，目的是表征该基因 在非肿瘤 TN 中与 TNBC 进行比较，并确定受其在癌症中表达影响的基因。这 实验方法是敲低样本中的MYBL1基因，进行微阵列分析 识别受过程差异影响的基因，然后进行进一步分析以验证 在此过程中确定的候选基因。作为概念证明，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富集，其中一些是新颖的。这一观察结果与 那些显示 MYBL1 和 MYBL2 共表达并证实 MYBL1 的能力 调节 MYBL2 的表达。当前研究中概述的实验应该扩大我们的研究范围 了解三阴性乳腺样本中的信号事件。对于目标 1，调查人员将 继续 TNBC 中 MYBL1 的敲低研究，并包括额外的 TNBC 和非肿瘤 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： 与三阴性癌症相比，非肿瘤中的 MYBL1 努力识别基因和过程 可能与肿瘤进展有关。由于 TNBC 具有异质性，本研究仅限于 B 亚型，MYBL1 表达样本。将鉴定可靠的特征基因，并进行途径分析 执行。假设 MYBL1 在不同组织中的表达会导致不同的结果 信号机制。将使用组织微阵列检查选择的差异表达基因 具有明确的受体状态和生存数据，并与 MYBL1 基因表达模式进行比较。为了 目标2，各种敲低样品的增殖能力和侵袭潜力将是 比较的。对于目标 3，未经处理的对照、shRNA 混杂对照和 每个细胞系的 MYBL1 shRNA 敲除制剂将通过软琼脂分析进行检查。