Targeting matrilysin and its impact on tumor growth in vivo:: The potential implications in breast cancer therapy
Targeting matrilysin and its impact on tumor growth in vivo:: The potential implications in breast cancer therapy
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DOI：
10.1158/1078-0432.ccr-05-0275
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发表时间：
2005-08-15
影响因子：
11.5
通讯作者：
Mansel, RE
中科院分区：
文献类型：
--
作者：
Jiang, WG;Davies, G;Mansel, RE
简介：Matrilysin (MMP-7) 是一种金属蛋白酶，参与细胞外基质的降解、侵袭和肿瘤进展。目前的研究检查了使用逆转录病毒核酶转基因靶向基质溶解素是否可能对乳腺癌细胞产生影响，并可能具有临床意义。实验设计：逆转录病毒锤头状核酶转基因被设计用于特异性靶向人类基质溶解素mRNA。乳腺癌细胞MDA-MB-231用逆转录病毒基质溶解素转基因或对照逆转录病毒转基因转染。在体外测试稳定转染的细胞的侵袭性和迁移特性。这些细胞还用于在无胸腺裸鼠中创建肿瘤模型，评估肿瘤的生长和基质溶解素的水平。此外，还对人类乳腺肿瘤队列中的基质溶素蛋白和 mRNA 水平进行了研究。 结果：反转录 PCR 表明，基质溶素的逆转录病毒锤头核酶转基因成功消除了 MDA-MB-231 中基质溶素的表达。与野生型（MDA-MB-231（WT）；26.2 +/- 6.2，P < 0.05）或对照转基因癌细胞相比，Matrilysin转基因转导的癌细胞（MDA-MB-231（Delta Matrilysin））表现出显着较低的侵袭程度（侵袭细胞数16.0 +/- 2.5） （MDA-MB-231（pRevTRE）；25.3 +/- 4.2，P < 0.01）。然而，细胞在体外的生长速度并未受到显着影响。在体内肿瘤模型中，与野生型 MDA-MB-231(WT) (1.46 +/- 0.04 cm(3)) 相比，MDA-MB-231(Delta Matrilysin) 肿瘤的免疫反应性 matrilysin 水平非常低，其生长速度明显较低（接种后 4 周为 0.24 +/- 0.03 cm(3)）。 MDA-MB-231(pRevTRE) (1.12 +/- 1.0 cm(3)) 肿瘤。在人类乳腺肿瘤中，与正常乳腺上皮相比，乳腺癌细胞对基质溶解素的染色密度明显更高。在高级别肿瘤以及中度和不良预后的患者中观察到最高水平的基质溶解素。最后，高水平的基质溶素与较差的长期存活率显着相关（P = 0.0143）。结论：在人类乳腺肿瘤中异常表达的基质溶素可以通过锤头核酶转基因从乳腺癌细胞中有效消除。基质溶解素的消除与低侵袭性和缓慢的肿瘤生长有关。综上所述，该研究表明，靶向基质溶解素可能具有重要的治疗意义。
Introduction: Matrilysin (MMP-7) is a metalloproteinase that is involved in the degradation of extracellular matrix, invasion, and tumor progression. The current study examined if targeting matrilysin using retroviral ribozyme transgenes may have an impact on breast cancer cells and may have clinical implications.Experimental Design: Retroviral hammerhead ribozyme transgenes were designed to specifically target human matrilysin mRNA. The breast cancer cell MDA-MB-231 was transfected with either a retroviral matrilysin transgene or a control retroviral transgene. Stably transfected cells were tested for their invasiveness and migratory properties in vitro. The cells were also used in creating a tumor model in athymic nude mice in which the growth of tumors and levels of matrilysin were assessed. In addition, levels of both protein and mRNA of matrilysin were investigated in a cohort of human breast tumors.Results: Expression of matrilysin in MDA-MB-231 was successfully eliminated by the retroviral hammerhead ribozyme transgene for matrilysin as revealed by reverse transcription-PCR. Matrilysin transgene-transduced cancer cells (MDA-MB-231(Delta Matrilysin)) exhibited a significantly lower degree of invasion (number of invading cells 16.0 +/- 2.5) compared with wild type (MDA-MB-231(WT); 26.2 +/- 6.2, P < 0.05) or control transgene-transcluced cancer cells (MDA-MB-231(pRevTRE); 25.3 +/- 4.2, P < 0.01). However, the rate of growth of the cells in vitro was not significantly affected. In the in vivo tumor model, MDA-MB-231(Delta Matrilysin) tumors, which had very low levels of immunoreactive matrilysin, grew at a significantly lower rate (0.24 +/- 0.03 cm(3), 4 weeks after inoculation) compared with the wild-type MDA-MB-231(WT) (1.46 +/- 0.04 cm(3)) and MDA-MB-231(pRevTRE) (1.12 +/- 1.0 cm(3)) tumors. In human breast tumors, breast cancer cells stained matrilysin at a significantly higher density, compared with normal mammary epithelium. The highest level of matrilysin was seen in high-grade tumors and that from patients with moderate and poor prognosis. Finally, high levels of matrilysin were significantly linked with a poor long-term survival (P = 0.0143).Conclusion: Matrilysin, which is aberrantly expressed in human breast tumors, can be effectively eliminated from breast cancer cells by way of hammerhead ribozyme transgene. Elimination of matrilysin is associated with low invasiveness and slow tumor growth. Taken together, the study suggests that targeting matrilysin may have important therapeutic implications.