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组织型基质金属蛋白酶抑制剂-2对白血病SHI-1细胞生长及体内浸润的影响 被引量:3

Overexpression of tissue inhibitor of metalloprotease-2 promotes proliferation and infiltration of human monocytic leukemia cells
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摘要 目的研究组织型基质金属蛋白酶抑制剂-2(TIMP-2)过表达对白血病 SHI-1细胞增殖、侵袭能力以及于裸鼠体内浸润、转移能力的影响。方法用流式细胞术检测 SHI-1细胞膜表面TIMP-2的表达。四甲基偶氮唑盐比色法(MTT 法)及与白血病患者骨髓基质细胞共培养跨人工基底膜侵袭实验检测高表达 TIMP-2的 SHI-1细胞体外增殖及侵袭能力的改变。经尾静脉接种进行预处理后的6周龄裸鼠,于接种后第30天处死部分裸鼠,检测 SHI-1细胞在各脏器中浸润生长情况。结果 SHI-1/TIMP-2细胞膜表面 TIMP-2的表达为99.3%±0.1%,显著高于 SHI-1/MSCV 细胞TIMP-2表达(85.9%±2.6%,P<0.05),平均荧光强度也提高2.06倍。SHI-1/TIMP-2细胞体外增殖能力及跨人工基底膜侵袭能力强于 SHI-1/MSCV。接种 SHI-1/TIMP-2细胞的裸鼠平均生存期为33.7 d,而接种 SHI-1/MSCV 细胞的裸鼠为40 d,两者之间平均生存期差异有统计学意义,体内浸润转移现象显著增强。结论膜表面高表达 TIMP-2的人单核细胞白血病细胞株 SHI-1/TIMP-2较 SHI-1/MSCV 细胞具有更强的增殖、浸润和转移能力。 Objective To investigate the functional role of human tissue inhibitor of metalloprotease (TIMP)-2 gene on the proliferation and infiltrating capability of human monocytic leukemic cells. Methods Human monocytic leukemic cells of the line SHI-1 were cultured and the TIMP-2 expression on the cell membrane was detected by flow cytometry (FCM). The SHI-1 cells were transfected with human TIMP-2 gene ( SHI-1/TIMP-2 cells) or blank vector ( SHI-1/MSCV cells). The TIMP-2 expressed on the surface of the cell membranes of SHI-1/TIMP-2 and SHI-1/MSCV cells was detected by FCM. The SHI-1/TIMP-2 and SHI-1/MSCV cells were inoculated into the 96-well plate, 24, 48, 72, and 96 hours later MMT method and ELISA were used, and the cell growth curve was drawn to detect the proliferation ability of the cells. Matrigel was put into the upper layer of Transwell chamber. Human bone marrow matrix cells (BMMC) of leukemia patient and SHI-1/TIMP-2 cells or SHI-1/MSCV cells were put into the upper layers as experimental groups, and SHI-1/TIMP-2 cells or SHI-1/MSCV cells only, without human BMMC, were put into the upper layers as control groups. 72 hours later blood cell counting plate was used to measure the number of cells migrating through Matrigel. SHI-1/TIMP-2 cells or SHI-1/MSCV cells were injected intravenously into pre-treated BALB/c nu/nu mice. Thirty days later 8 mice from each group were killed to observe the tumorigenesis in the organs, especially the central nervous system leukemia (CNL). The survival of the other mice was observed. Results The expression level of TIMP-2 on the cell membrane of the SHI-1/TIMP-2 cells was 99. 3%±0. 1% , significantly higher than that of the SHI-1/MSCV cells (85.9%±2.6% , P 〈0. 05). The A values 24, 48, 72, and 96 hours later of the SHI-1/TIMP-2 cells were 0. 34±0.05, 0.6± 0.05, 0.97± 0.12, and 1.28±0.06 respectively, all significantly higher than those of the SHI-1/MSCV cells (0.28±0.03, 0. 36±0. 03, 0. 54±0.09, and 0. 99±0. 03 respectively, all P 〈 0.05 ). The SHI-1/TIMP-2 ceils and SHI-1/MSCV ceils only could not migrate through Matrigel basically. 24-48 hours after co-cultivation Shi-1 cells began to appear in the lower layer of Transwell chambers, 72 hours later the trans-Matrigel SHI-1/TIMP-2 cells accounted for 24. 7%± 6.9% of the inoculated SHI-1/TIMP-2 cells, a proportion significantly higher than that in the case of the SHI-1/MSCV cells ( 12%±1.4%, P 〈 0. 05 ). 24 days after the inoculation the mean body weight of the mice inoculated with SHI-1/TIMP-2 cells was 21.5 g±0.4 g, significantly higher than that of the mice inoculated with SHI- 1/MSCV cells ( 17.4 g±0. 6 g, P 〈 0. 01 ). The mice inoculated with SHI-1/TIMP-2 cells showed much more tumors in different organs and much more severe infiltration in the CNS in comparison with the mice inoculated with SHI-1/MSCV cells The mean survival time of the mice inoculated with the SHI-1/TIMP-2 cells was 33.7 days, significantly shorter than that of the mice inoculated with SHI-1/MSCV cells (40 days). Conclusion TIMP-2 expressed on the cell membrane is critical to promote the proliferation and infiltration of SHI-1 cells.
出处 《中华医学杂志》 CAS CSCD 北大核心 2006年第34期2409-2412,共4页 National Medical Journal of China
基金 江苏省卫生厅基金(H200327)
关键词 白血病 单核细胞 急性 基质金属蛋白酶 小鼠 肿瘤转移 Leukemia, monocytic, acute Matrix metalloproteinases Mice, nude Neoplasm metastasis
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