EFFECT OF VITAMIN D ON GENE EXPRESSION IN MCF-7 CELLS

维生素 D 对 MCF-7 细胞基因表达的影响

• 批准号: 2796290
• 负责人: ROBERT R BURAS
• 金额: $ 8.91万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2796290
• 关键词: 1,25 dihydroxycholecalciferol; MCF7 cell; RNase protection assay; antineoplastics; breast neoplasms; cell cycle; cell differentiation; colon neoplasms; complementary DNA; cycloheximide; gel mobility shift assay; gene induction /repression; genetic promoter element; leukemia; melanoma; molecular cloning; neoplasm /cancer pharmacology; northern blottings; nucleic acid sequence; nutrition related tag; plasmids; polymerase chain reaction; prostate neoplasms; vitamin D; vitamin analog

The candidate for this Clinical Investigator Award, Dr. Robert Buras, is a surgical oncologist with six years of clinical training and four years of research training. He currently devotes 40% of his effort to research and 60% to clinical and teaching activities. This award would enable Dr. Buras to increase his research effort from 40% to 80%. He would use this opportunity to study the mechanisms through which 1,25-(OH)2-vitamin D3(1,25-D2) and vitamin D analogs inhibit breast cancer cell growth. The proposed studies will analyze differential gene expression in MCF-7 breast cancer cells treated with 1,25-D3 and Ro24-5531. MCF-7 cells express vitamin D receptors (vDR) and demonstrate significant growth inhibition by 10-8M 1,25-D3 or Ro24-5531. Ro24-5531, a non-hypercalcemic vitamin D analog, decreases the incidence of breast tumors in a rat carcinogenesis model. MCF-7 cells will be grown in standard media, in media depleted of vitamin D and in media supplemented with 1,25-D3 or Ro24-5531. Differential gene expression will be analyzed by PCR-based differential display. Differentially-expressed genes will be cloned, sequenced and analyzed for homology with known proteins or functional groups to determine possible significance to the antiproliferative effect of 1,25-D3 and vitamin D analogs. The cellular distribution and timing of expression of regulated gene products will be studied using anti- protein antibodies. The effects of regulated proteins on cell growth, differentiation, invasiveness and progression through the cell cycle will be studied. The promoter region of genes which mediate the antiproliferative effects of 1,25-D3 or Ro24-5531 will be analyzed to determined mechanisms of transcriptional regulation. These studies will improve our understanding of the mechanisms through which 1,25-D3 and Ro24-5531 regulate cancer cell growth and differentiation. This insight may suggest new strategies for the prevention and treatment of breast cancer. Dr. Buras' research will be conducted under the direction of Dr. Marc Lippman, Director of the Lumbardi Cancer Center (LCC). The LCC has the facilities necessary for Dr. Buras to successfully complete this research. Numerous senior staff of the LCC, including many established breast cancer researchers, will be available to Dr. Buras for consultation. At the conclusion of this award, Dr. Buras will have acquired the experience and critical thinking necessary to direct an independent research program.

本次临床研究者奖的候选人 Robert Buras 博士是 一名接受过六年临床培训和四年的肿瘤外科医师 研究培训。他目前将40%的精力投入到研究上 60%用于临床和教学活动。该奖项将使博士能够 Buras 将他的研究工作量从 40% 增加到 80%。他会用这个 有机会研究 1,25-(OH)2-维生素的机制 D3(1,25-D2) 和维生素 D 类似物可抑制乳腺癌细胞生长。 拟议的研究将分析 MCF-7 中的差异基因表达 用 1,25-D3 和 Ro24-5531 处理的乳腺癌细胞。 MCF-7细胞 表达维生素 D 受体 (vDR) 并表现出显着的生长 10-8M 1,25-D3 或 Ro24-5531 抑制。 Ro24-5531，一种非高钙血症 维生素 D 类似物可降低大鼠乳腺肿瘤的发生率 致癌模型。 MCF-7 细胞将在标准培养基中生长， 缺乏维生素 D 的培养基和补充有 1,25-D3 或 罗24-5531。将通过基于 PCR 的方法分析差异基因表达 差分显示。差异表达基因将被克隆， 测序并分析与已知蛋白质或功能性蛋白质的同源性 组以确定抗增殖作用的可能意义 1,25-D3 和维生素 D 类似物。细胞分布和时间 将使用抗来研究受调节基因产物的表达 蛋白质抗体。调节蛋白对细胞生长的影响， 细胞周期中的分化、侵袭和进展将 被研究。介导基因的启动子区域 将分析 1,25-D3 或 Ro24-5531 的抗增殖作用 确定的转录调控机制。 这些研究将通过以下方式增进我们对机制的理解： 其中 1,25-D3 和 Ro24-5531 调节癌细胞生长 差异化。这一见解可能会为以下领域提出新的策略： 预防和治疗乳腺癌。 Buras 博士的研究将在 Marc 博士的指导下进行 Lippman，伦巴迪癌症中心 (LCC) 主任。低成本航司拥有 Buras 博士成功完成此任务所需的设施 研究。 LCC 拥有众多高级职员，其中包括许多资深人士 乳腺癌研究人员将向 Buras 博士提供帮助 咨询。在该奖项结束时，Buras 博士将获得 获得了指导所需的经验和批判性思维 独立研究计划。