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: 10618894
• 负责人: Audrey Player
• 金额: $ 11.42万
• 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/10618894
• 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; MYBL1 gene; MYBL2 gene; Malignant Neoplasms; Mammary Neoplasms; Microarray Analysis; Oncogenes; Pathway Analysis; Patients; Pattern; Play; Preparation; Principal Investigator; Procedures; Process; Proliferating; Property; Proto-Oncogenes; Publishing; Regulation; Research Personnel; Role; Sampling; Signal Transduction; Signaling Protein; Statistical Data Interpretation; Testing; Tissue Microarray; Tissues; Transcription Coactivator; Uterine Fibroids; Validation; aggressive breast cancer; 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 cancer; 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.

MYBL 1基因敲低在非肿瘤性和三阴性乳腺癌中的比较分析 细胞 项目摘要 MYBL 1基因属于MYB基因家族，包括c-MYB和MYBL 2。的基因 是作为参与增殖的强转录激活因子起作用的原癌基因， 分化和细胞周期信号传导过程，所有这些都是肿瘤进展中的关键事件。研究 表明截短的c-MYB是一种致癌基因，因此，该基因被研究为治疗靶点， 乳腺癌对MYBL 1和MYBL 2的了解较少。两者在细胞周期中起重要作用 在不同的癌症，包括乳腺癌中过度表达。的已发表结果 Player实验室显示MYBL 1在三阴性乳腺癌亚群中过表达， （TNBC）。在一个单独的综合监督网络分析管腔乳腺癌，MYBL 1 显示过表达与疾病预后差相关。根据这些研究，校长 研究者（PI）实验室选择研究MYBL 1基因，并对该基因的特征感兴趣 与TNBC相比，在非肿瘤TN中的表达，并鉴定受其在癌症中表达影响的基因。的 实验方法是敲除样品中的MYBL 1基因，进行微阵列分析， 确定受过程影响的差异基因，然后进行进一步分析， 在这个过程中发现的候选基因。作为概念证明，MYBL 1基因在细胞内表达下调， 在TNBC细胞系中。受MYBL 1表达降低影响的候选基因包括MYBL 2和MYBL 3。 细胞周期信号基因的富集，其中一些是新的。这一观察结果与 那些显示MYBL 1和MYBL 2共表达并证实MYBL 1能够 调节MYBL 2的表达。目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，研究人员将 继续在TNBC中敲除MYBL 1的研究，并包括额外的TNBC和非肿瘤TN 样品作为比较。这些数据将允许PI通过以下方式识别不同的调节模式： 与三阴性癌症相比，MYBL 1在非肿瘤中的作用， 可能与肿瘤进展有关。由于TNBC是异质性的，因此本研究仅限于 亚型B，MYBL 1表达样品。将鉴定可靠的标记基因，并进行途径分析 执行。假设MYBL 1在不同组织中的表达导致了不同的细胞凋亡。 信号机制。选择差异表达的基因将使用组织微阵列进行检查 具有确定的受体状态和存活数据，并与MYBL 1基因表达模式进行比较。为 目的2，将各种敲除样品的增殖能力和侵袭潜力进行比较。 比较了对于目标3，未处理的对照、shRNA乱序对照和未处理的对照的致瘤特性。 将通过软琼脂分析检查各细胞系的MYBL 1 shRNA敲减制备物。