MOLECULAR ANALYSIS OF HEPATOMA CELL MIGRATION FACTOR

肝癌细胞迁移因子的分子分析

• 批准号: 3201705
• 负责人: YU YAMAGUCHI
• 金额: $ 21.23万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3201705
• 关键词: cell differentiation; cell migration; cytokine; hepatocellular carcinoma; human tissue; immunocytochemistry; laboratory mouse; laboratory rabbit; laboratory rat; molecular cloning; neoplastic growth; protein purification; transfection

Cell locomotion is essential for tumor cells to invade into surrounding tissues and achieve metastasis. Soluble factors that enhance cellular motility have been implicated in the process. We have found an activity in conditioned media of human fibroblasts that stimulates the motility of hepatoma cells. The long-term objective of this proposal is to characterize this novel factor, tentatively termed "hepatoma cell migration factor" (HMF). HMF disperses tightly packed colonies of well-differentiated hepatoma cells into single cells and promotes the migration of the cells out of colonies. The dispersed cells spread extensively and assume a fibroblastic, dedifferentiated morphology. These phenotypical changes are induced with less than 1 ng/ml of partially purified HMF. The estimated molecular weight (60 kDa), the narrow range of target cell specificity toward hepatocytes/hepatoma cells, and other biological and biochemical data suggest that HMF is distinct from other motility factors and cytokines known to affect cell morphology and motility. Thus, our working hypothesis is that HMF is a novel cytokine-type molecule which regulates the migration and differentiation of hepatocytes/hepatoma cells and possibly the invasion and metastasis of hepatoma in vivo. To examine the biological roles of HMF, we will 1) purify HMF and obtain its amino acid sequences; 2) molecularly clone and sequence HMF cDNA; 3) investigate the tissue distribution of HMF by immunohistochemical staining with specific antibody probes; and 4) characterize the biological effects of HMF on the differentiation, migration and invasion of hepatocytes/hepatoma cells by using exogenously added HMF and by gene transfer of HMF cDNA into cells. These studies will contribute to our understanding of the molecular mechanisms of the invasion and metastasis of hepatoma in vivo and define a role for HMF in these processes.

细胞运动对于肿瘤细胞侵入周围环境至关重要 组织并实现转移。 增强细胞的可溶性因子 运动性参与了这个过程。 我们找到了一个活动 在人成纤维细胞的条件培养基中刺激运动 肝癌细胞。 该提案的长期目标是 描述了这种新因子，暂时称为“肝癌细胞” 迁移因子”（HMF）。 HMF 分散紧密堆积的高分化肝癌集落 细胞变成单个细胞并促进细胞迁移出 殖民地。 分散的细胞广泛分布并呈现出 成纤维细胞、去分化形态。 这些表型变化 用低于 1 ng/ml 的部分纯化的 HMF 进行诱导。 这 估计分子量（60 kDa），靶细胞范围窄 对肝细胞/肝癌细胞和其他生物和细胞的特异性 生化数据表明 HMF 与其他运动因子不同 和已知影响细胞形态和运动的细胞因子。 因此，我们的 工作假设是 HMF 是一种新型细胞因子型分子， 调节肝细胞/肝癌细胞的迁移和分化 也可能是肝癌在体内的侵袭和转移。 为了检查 HMF 的生物学作用，我们将 1) 纯化 HMF 并获得 其氨基酸序列； 2) HMF cDNA分子克隆及测序； 3） 通过免疫组化研究 HMF 的组织分布 用特异性抗体探针染色； 4）表征 HMF对分化、迁移和侵袭的生物学作用 通过使用外源添加的 HMF 和基因对肝细胞/肝癌细胞 将 HMF cDNA 转移到细胞中。 这些研究将有助于我们 了解侵袭和转移的分子机制 体内肝癌的研究并定义了 HMF 在这些过程中的作用。