TIMP-1 EXPRESSION BY NORMAL LYMPHOCYTES AND IN LYMPHOID NEOPLASMS

正常淋巴细胞和淋巴肿瘤中的 TIMP-1 表达

• 批准号: 3774420
• 负责人: M STETLER-STEVENSON
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3774420
• 关键词: B lymphocyte; Burkitt's lymphoma; RNA; T lymphocyte; athymic mouse; cell cycle; clone cells; collagenase; disease /disorder model; enzyme activity; enzyme mechanism; gene expression; human tissue; metalloenzyme; neoplasm /cancer genetics; neoplasm /cancer invasiveness; northern blottings; protein purification; tissue inhibitor of metalloproteinases; transfection

We have studied the expression of TIMP-1 in a series of Burkitt cell lines. Burkitt's lymphoma, common in AIDS patients, is an aggressive neoplasm without effective therapy in the immunocompromised host. Northern blot analysis reveals that TIMP-1 is expressed at the RNA level by 4 out of 9 lines studied. Cell lines from both African and American Burkitt's express TIMP-1, as do Burkitt cells with t(8;14) and t(8;22) translocations. All of the Burkitt cell lines studied with demonstrable TIMP-1 RNA transcripts secreted TIMP-1 protein (as determined by Western blot analysis) into the culture media. TIMP-1 expression does not correlate with EBV involvement, p53 mutations, or levels of myc expression in these lines. The cell lines that express TIMP-1 have a higher rate of cell proliferation in vitro compared to those negative for TIMP-1 transcripts. We are purifying TIMP-1 protein and intend to study the effect of TIMP-1 on growth kinetics in vitro. The TIMP-1 expressing Burkitt cell lines are also more tumorigenic and invasive in the nude mouse model. Invasion of skin and nerve is seen in only in TIMP-1 expressing lines. This aggressive behavior may be due to high growth rate, enhanced invasive behavior or a combination of the two. We are currently studying four cell lines (two expressing TIMP-1 and two negative) in invasion assays to assess invasive behavior. We plan to infect TIMP-1 negative cell lines with a retroviral construct containing TIMP-1 to cause TIMP-1 expression in order to study the effect of TIMP-1 on growth rate and invasiveness. In four of the Burkitt cell lines studied, Gelatinase B activity can be demonstrated by zymogram but not by Northern blot analysis. TIMP-1 and Gelatinase B coexpression is seen in only one cell line, with very low enzyme expression noted. The remainder of the Gelatinase B activity was demonstrated in TIMP-1 negative cell lines. Gelatinase A expression was not observed in any Burkitt cell lines. If TIMP-1 functions in Burkitt's lymphoma by blocking gelatinase activity, we would have expected to be able to demonstrate gelatinase expression by Northern blot or zymogram in the TIMP-1 positive cell lines. As gelatinase expression may occur in these cells in response to the extracellular matrix, we are studying the expression of Gelatinase A and Gelatinase B in Burkitt cell tumors grown in nude mice.

我们研究了TIMP-1在一系列Burkitt细胞中的表达 线。伯基特淋巴瘤常见于艾滋病患者，是一种侵袭性淋巴瘤 免疫功能低下的宿主中未经有效治疗的肿瘤。 Northern blot 分析显示 TIMP-1 在 RNA 水平表达 研究了 9 个品系中的 4 个。 来自非洲和美洲的细胞系 Burkitt 细胞表达 TIMP-1，t(8;14) 和 t(8;22) 的 Burkitt 细胞也表达 TIMP-1 易位。 所有 Burkitt 细胞系的研究均具有可证明的 TIMP-1 RNA 转录本分泌 TIMP-1 蛋白（Western 测定） 印迹分析）到培养基中。 TIMP-1 表达不 与 EBV 参与、p53 突变或 myc 表达水平相关 在这些行中。 表达 TIMP-1 的细胞系具有更高的比率 与 TIMP-1 阴性细胞相比，体外细胞增殖情况 成绩单。 我们正在纯化 TIMP-1 蛋白并打算研究 TIMP-1 对体外生长动力学的影响。 TIMP-1表达 伯基特细胞系在裸体中也更具致瘤性和侵袭性 鼠标模型。 仅在 TIMP-1 中可见皮肤和神经的侵袭 表达线条。 这种攻击行为可能是由于高生长所致 率、增强的侵入行为或两者的组合。 我们是 目前正在研究四种细胞系（两种表达 TIMP-1，两种表达 阴性）在入侵测定中评估入侵行为。 我们计划 用含有以下成分的逆转录病毒构建体感染 TIMP-1 阴性细胞系 TIMP-1引起TIMP-1表达以研究TIMP-1的作用 关于生长速度和侵袭力。 在所研究的四种 Burkitt 细胞系中，明胶酶 B 活性可 由酶谱证明，但未由 Northern 印迹分析证明。 TIMP-1 和 仅在一种细胞系中观察到明胶酶 B 共表达，且表达量非常低 注意到酶的表达。 其余的明胶酶 B 活性为 在 TIMP-1 阴性细胞系中得到证实。 明胶酶 A 表达为 在任何 Burkitt 细胞系中均未观察到。 如果 TIMP-1 在伯基特氏病中发挥作用 通过阻断明胶酶活性，我们预计淋巴瘤 能够通过 Northern blot 或酶谱证明明胶酶的表达 TIMP-1 阳性细胞系。 由于明胶酶表达可能发生在 这些细胞对细胞外基质做出反应，我们正在研究 明胶酶 A 和明胶酶 B 在伯基特细胞肿瘤中的表达 在裸鼠中。