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: 2895141
• 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 2001-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2895141
• 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.

这一临床研究人员奖的候选人罗伯特·伯拉斯博士是 有六年临床训练和四年经验的外科肿瘤学家 进行研究训练。他目前将40%的精力投入到研究中 60%用于临床和教学活动。这一奖项将使Dr. Buras将他的研究努力从40%增加到80%。他会用这个 研究1，25-(OH)2-维生素作用机制的机会 D3(1，25-D2)和维生素D类似物抑制乳腺癌细胞生长。 拟议的研究将分析MCF-7细胞的差异基因表达 用1，25-D3和Ro24-5531处理乳腺癌细胞。MCF-7细胞 表达维生素D受体(VDR)并表现出显著的生长 10-8M、1，25-D3或Ro24-5531的抑制作用。Ro24-5531，一种非高钙血症 维生素D类似物，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的培养液和添加1，25-D3或 Ro24-5531。差异基因表达将通过基于聚合酶链式反应的分析 差异显示。差异表达的基因将被克隆， 测序并分析与已知蛋白质或功能的同源性 小组确定对抗增殖作用的可能意义 1，25-D3和维生素D类似物。蜂窝分布和时序 对受调控基因产物的表达的研究将使用抗 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。基因的启动子区域，它介导了 1，25-D3或Ro24-5531的抗增殖作用将被分析为 已确定的转录调控机制。 这些研究将通过以下方式提高我们对这些机制的理解 其中1，25-D3和Ro24-5531调节癌细胞生长和 差异化。这一洞察力可能会为 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 伦巴迪癌症中心(LCC)主任利普曼说。LCC拥有 Buras博士成功完成这项工作所需的设施 研究。LCC的许多高级工作人员，包括许多已建立的 乳腺癌研究人员，将向Buras博士提供 咨询。在这个奖项结束时，Buras博士将拥有 获得必要的经验和批判性思维来指导 独立研究计划。