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: 3752120
• 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/3752120
• 关键词: B lymphocyte; Burkitt's lymphoma; CD antigens; Retroviridae; T lymphocyte; apoptosis; athymic mouse; cold temperature; enzyme mechanism; gene expression; human tissue; metalloenzyme; neoplasm /cancer genetics; neoplasm /cancer invasiveness; neoplastic growth; tissue /cell culture; 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没有有效治疗的肿瘤。 北方印迹分析显示TIMP-1在RNA水平表达 研究的9条线中有4条。 非洲和美洲的细胞系 Burkitt's表达TIMP-1，t（8;14）和t（8;22）的Burkitt细胞也表达TIMP-1 易位 研究的所有Burkitt细胞系都具有可证明的 TIMP-1 RNA转录物分泌TIMP-1蛋白（如通过Western印迹法测定）。 印迹分析）到培养基中。 TIMP-1表达不 与EBV感染、p53突变或myc表达水平相关 在这些线。 表达TIMP-1的细胞系具有更高的凋亡率。 与TIMP-1阴性的细胞相比，体外细胞增殖 成绩单 我们正在纯化TIMP-1蛋白，并打算研究TIMP-1蛋白的作用。 TIMP-1对体外生长动力学的影响。 TIMP-1表达 Burkitt细胞系在裸细胞中也更具致瘤性和侵袭性 小鼠模型 仅在TIMP-1中可见皮肤和神经浸润 表达线条。 这种攻击性行为可能是由于高增长 率，增强的侵入行为或两者的组合。 我们 目前正在研究四种细胞系（两种表达TIMP-1，两种表达TIMP-2）， 阴性）以评估侵袭行为。 我们计划 用逆转录病毒构建体感染TIMP-1阴性细胞系，所述构建体含有 为了研究TIMP-1的作用， 生长速度和侵袭性。 在所研究的四种伯基特细胞系中，明胶酶B活性可以被检测到。 通过酶谱而不是通过北方印迹分析证实。 TIMP-1和 明胶酶B共表达仅见于一个细胞系， 酶的表达。 明胶酶B活性的其余部分为 在TIMP-1阴性细胞系中证实。 明胶酶A表达为 在任何伯基特细胞系中均未观察到。 如果TIMP-1在Burkitt's 淋巴瘤通过阻断明胶酶活性，我们本来预计 能够通过北方印迹或酶谱证明明胶酶表达， TIMP-1阳性细胞株。 由于明胶酶表达可能发生在 这些细胞对细胞外基质的反应，我们正在研究 明胶酶A和明胶酶B在Burkitt细胞瘤中的表达 在裸鼠中。