Taxane Resistance in Breast and Ovarian Cancer Cells

乳腺癌和卵巢癌细胞中的紫杉烷耐药性

• 批准号: 8074492
• 负责人: BRANIMIR I SIKIC
• 金额: $ 28.88万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8074492
• 关键词: ABCB1 gene; Accounting; Adverse effects; Affect; Antibodies; Apoptosis; Apoptosis Regulator; Apoptotic; Binding; Breast; Breast Cancer Cell; Breast Carcinoma; Cancer cell line; Candidate Disease Gene; Cell Line; Cell model; Cells; Cetuximab; Clinical; Clinical Markers; Collaborations; Cooperative Human Tissue Network; Data; Drug resistance; EGFR inhibition; Epidermal Growth Factor Receptor; Gefitinib; Gene Expression Profiling; Genes; Genetic; Genetic Markers; Genome; Genomics; Growth; Immunoblotting; Jordan; Kinetics; Lung; MAP4; MCF7 cell; Malignant Neoplasms; Malignant neoplasm of ovary; Mediator of activation protein; Microarray Analysis; Microtubule-Associated Proteins; Microtubules; Minority; Molecular Target; Mutation; Ovary; P-Glycoprotein; Paclitaxel; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphorylation; Prostate; Protein Isoforms; Proteins; RNA Interference; Regulation; Relative (related person); Research Personnel; Resistance; SDZ-PSC-833; Small Interfering RNA; Specimen; Taxane Compound; Technology; Tetanus Helper Peptide; Tetracyclines; Tubulin; Tyrosine Kinase Inhibitor; Variant; cancer cell; cell growth; chemotherapeutic agent; docetaxel; drug sensitivity; inhibitor/antagonist; insight; interest; kinase inhibitor; malignant breast neoplasm; novel; overexpression; programs; resistance factors; resistance mechanism; response; tau Proteins; taxane; tumor; vector

DESCRIPTION (provided by applicant): This proposal will study non-MDR1 genetic and cellular determinants of taxane therapy in breast and ovarian cancers. We have established 17 taxane-resistant variants from 4 breast and 6 ovarian cancer cell lines, and we hypothesize that these diverse variants may yield insight into novel mechanisms of resistance to taxanes. The specific aims are: 1. To determine tubulin isoform content and potential alterations in microtubule dynamic instability. Our analyses of the expression of the six classes of (3- tubulin isoforms reveal altered p-tubulin content in many of the taxane-resistant variants. Microtubule assembly in these cells will be assessed in collaboration with Dr. Mary Ann Jordan. 2. To study the effects of altered tubulin and microtubule associated protein genes on drug sensitivity. Several of the variants have down-regulated MAP4 and up-regulated MAP Tau, and we hypothesize that these alterations may have an adverse effect on taxane sensitivity by altering taxane binding. We plan to use siRNA technology to study the effects of these genes and specific tubulin isoforms, and inducible vectors utilizing tetracycline regulation will be used to transfect genes of interest into parental cells. 3. To explore regulators of apoptosis (e.g. Bcl-2, Bcl-XL, Akt) and cell growth (e.g. EGFR-1, her2) as determinants of response to taxanes. Several of the non-P-gp variants possess altered expression of apoptotic regulators. We will use antisense and RNAi approaches to study the effects of inhibition of these regulators on cellular sensitivity to taxanes. We will also determine the effects of inhibition of growth pathways on taxane sensitivity by inhibiting EGFR-1, her2, Akt and other genes using kinase inhibitors. 4. To identify novel genetic markers to predict resistance to taxanes. Genetic (array CGH) and genomic (microarray gene expression profiling) approaches will be used to search for novel genetic markers for the prediction of sensitivity and resistance to taxane therapy. Candidate genes will be validated by PCR, immunoblotting, and tet-regulated expression using an MCF-7 Flp-ln line which we have made. 5. To study taxane resistance in clinical specimens of breast and ovarian cancer. Whole genome profiling of tumor specimens from breast and ovarian cancers from the NCI Cooperative Human Tissue Network, annotated for responsiveness to taxane therapy, will be performed. Expression of resistance factors identified in cellular models will also be validated in these specimens. In summary, this project will use a large set of unique models of cellular resistance to taxanes in ovarian and breast cancer cell lines and clinical specimens obtained from patients treated with taxanes to: (1) study the relevance of known resistance mechanisms to these drugs, (2) explore new mechanisms, and (3) develop clinical markers for response.

描述（由申请人提供）：该提案将研究乳腺癌和卵巢癌中紫杉烷治疗的非MDR1遗传和细胞决定因素。我们已经建立了来自4种乳腺癌和6种卵巢癌细胞系的17种抗紫杉烷的变体，我们假设这些多样的变体可能会深入了解对紫杉烷耐药性的新型机制。具体目的是：1。确定微管动态不稳定性的微管蛋白同工型含量和潜在变化。我们对六类（3-微管蛋白亚型的表达的分析揭示了许多耐紫杉烷耐药性变体中的p微管蛋白含量变化。这些细胞中的微管组装将与玛丽·安·乔丹（Mary Ann Jordan）合作进行评估。 tau，我们假设这些变化可能会通过改变紫杉烷结合对紫杉烷的敏感性产生不利影响，我们计划使用这些基因的效果和特定的小管蛋白同工型，并使用四环素调节的诱导媒介将其用于转移利益的基因。 BCl-XL，AKT）和细胞生长（例如EGFR-1，HER2）作为对群群的反应的决定因素。我们还将通过使用激酶抑制剂抑制EGFR-1，HER2，AKT和其他基因来确定抑制生长途径对紫杉烷敏感性的影响。 4。确定新的遗传标记以预测对紫杉烷的抵抗力。遗传（阵列CGH）和基因组（微阵列基因表达分析）方法将用于寻找新的遗传标记，以预测敏感性和对紫杉烷治疗的抗性。候选基因将通过我们制作的MCF-7 FLP-LN系列通过PCR，免疫印迹和TET调节的表达来验证。 5。研究乳腺癌和卵巢癌临床标本中的紫杉烷耐药性。将进行对乳腺癌和卵巢癌的肿瘤标本的全基因组分析，从NCI合作的人体组织网络注释，以应对紫杉烷治疗的反应。在这些标本中也将验证细胞模型中鉴定的电阻因子的表达。总而言之，该项目将使用卵巢癌和乳腺癌细胞系中紫杉烷抗性的各种独特模型以及从接受紫杉烷治疗的患者获得的临床标本的抗紫杉烷的性能：

项目成果

Expression and silencing of the microtubule-associated protein Tau in breast cancer cells.

• DOI: 10.1158/1535-7163.mct-10-0780
• 发表时间: 2010-11
• 期刊: Molecular cancer therapeutics
• 影响因子: 5.7
• 作者: Spicakova T;O'Brien MM;Duran GE;Sweet-Cordero A;Sikic BI
• 通讯作者: Sikic BI

Decreased levels of baseline and drug-induced tubulin polymerisation are hallmarks of resistance to taxanes in ovarian cancer cells and are associated with epithelial-to-mesenchymal transition.

• DOI: 10.1038/bjc.2017.102
• 发表时间: 2017-05-09
• 期刊: British journal of cancer
• 影响因子: 8.8
• 作者: Duran GE;Wang YC;Moisan F;Francisco EB;Sikic BI
• 通讯作者: Sikic BI

Single Gene Prognostic Biomarkers in Ovarian Cancer: A Meta-Analysis.

• DOI: 10.1371/journal.pone.0149183
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Willis S;Villalobos VM;Gevaert O;Abramovitz M;Williams C;Sikic BI;Leyland-Jones B
• 通讯作者: Leyland-Jones B