VITAMIN D EFFECT 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: 2106548
• 负责人: ROBERT R BURAS
• 金额: $ 7.83万
• 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/2106548
• 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布拉斯博士是 一位接受过六年临床训练的外科肿瘤学家， 研究训练。他目前将40%的精力用于研究 60%用于临床和教学活动。该奖项将使博士。 布拉斯将他的研究工作从40%增加到80%。他会用这个 有机会研究1，25-（OH）2-维生素 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中的差异基因表达 用1，25-D3和Ro 24 -5531处理的乳腺癌细胞。MCF-7细胞 表达维生素D受体（vDR），并表现出显著的生长 10- 8 M 1，25-D3或Ro 24 -5531抑制。Ro 24 -5531，非高钙血症 维生素D类似物，降低大鼠乳腺肿瘤的发病率 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 Ro24-5531差异基因表达将通过基于PCR的 差异显示将克隆差异表达的基因， 测序并分析与已知蛋白质或功能蛋白质的同源性 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- 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。介导细胞凋亡的基因的启动子区是一个启动子区， 将分析1，25-D3或Ro 24 -5531的抗增殖作用， 确定转录调控机制。 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Ro 24 -5531调节癌细胞生长， 分化这一见解可能会提出新的战略， 乳腺癌的预防和治疗。 布拉斯博士的研究将在马克博士的指导下进行 Lumbardi癌症中心（LCC）主任Lippman说。LCC拥有 布拉斯博士成功完成这项工作所需的设施 research.地方合同委员会的许多高级工作人员，包括许多常设 乳腺癌研究人员，将提供给博士布拉斯， 协商在这个奖项结束时，布拉斯博士将有 获得了必要的经验和批判性思维来指导一个 独立研究计划。