Effect of matrix metalloproteinase inhibitor batimastat on breast cancer regrowth and metastasis in athymic mice.
Effect of matrix metalloproteinase inhibitor batimastat on breast cancer regrowth and metastasis in athymic mice.
复制标题
基质金属蛋白酶抑制剂巴马司他对无胸腺小鼠乳腺癌再生和转移的影响。
DOI：
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
R. Fife
中科院分区：
文献类型：
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作者：
G. Sledge;M. Qulali;R. Goulet;E. Bone;R. Fife
背景
基质金属蛋白酶(MMPs)通过介导细胞外基质成分的降解参与肿瘤的侵袭和转移。因此，这些酶在抗癌治疗的发展中是很有希望的靶点。巴替马斯特(([(4-N-hydroxyamino)-2R-isobutyl-3S-(thienyl-thiomethyl)succinyl]-L-苯丙氨酸-N-甲酰胺)是一种新型的抑制基质金属蛋白酶活性的药物。
目的
我们询问，在初次肿瘤切除后给予巴替马斯特作为辅助治疗，是否可以抑制人乳腺癌异种移植模型中局部区域肿瘤的再生长和肺转移的形成。我们还探索了batimastat对乳腺癌细胞活性和特定信使RNAs(MRNAs)积累的可能影响。
方法
体外培养的人乳腺癌细胞株MDA-MB-435经0.1~10.0微米的巴替马斯特作用6d后，进行活细胞计数。直接与培养的MDA-MB-435细胞结合或释放到其培养液中的胶原酶活性在用药1天和3天(药物范围为0.2-2.0微米)后用明胶酶谱检测。裸鼠在切除原发于乳腺脂肪垫的肿瘤后，每天腹腔注射巴替马斯特(30 mg/kg体重)，计算肿瘤再生长的体积和肺转移的数目和体积，并用抗内皮细胞抗原CD31的抗体进行免疫组织化学分析。用RNA斑点印迹和互补探针杂交的方法检测贝替马斯特对培养细胞、原发肿瘤和肿瘤再生过程中编码特异性MMPs和金属蛋白酶组织抑制因子-2(TIMP-2)的mRNAs积聚的影响。数据分析采用线性回归分析、t检验和卡方分析。
结果
培养的MDA-MB-435细胞的活力不受巴替马斯特处理的影响；然而，这些细胞释放的72kd和92kd胶原酶的测量活性在巴替马斯特处理后降低。腹腔注射巴替马斯特显著抑制裸鼠MDA-MB-435肿瘤的局部再生长(与对照组比较，P=.035)，减少肺转移发生率(P<；0.05)、肺转移数目(P=0.0001)和肺转移体积(P=0.0001)。巴替马斯特治疗不影响细胞内基质金属蛋白酶或TIMP-2mRNAs的水平。
结论
巴替马斯特抑制裸鼠移植瘤模型中人乳腺癌的再生长和转移。Batimastat抑制活性的潜在机制不包括直接细胞毒性或改变基质金属蛋白酶或TIMP的mRNA水平。
BACKGROUND
Matrix metalloproteinases (MMPs) are involved in the invasion and metastasis of human cancers by mediating the degradation of extracellular matrix components. Therefore, these enzymes constitute promising targets in the development of anticancer therapies. Batimastat ([(4-N-hydroxyamino)-2R-isobutyl-3S-(thienyl-thiomethyl)succinyl]-L- phenyl-alanine-N-methylamide) is one of a new class of agents designed to inhibit MMP activity.
PURPOSE
We asked whether batimastat, given as adjuvant therapy after primary tumor resection, could inhibit local-regional tumor regrowth and the formation of lung metastases in a human breast cancer xenograft model. We also explored possible effects of batimastat on breast cancer cell viability and on the accumulation of specific messenger RNAs (mRNAs).
METHODS
Human MDA-MB-435 breast cancer cells were treated in vitro for 6 days with batimastat at concentrations ranging from 0.1 to 10.0 microM, and then viable cell counts were performed. The activity of collagenases, directly associated with cultured MDA-MB-435 cells or released into their culture fluids, was assessed by gelatin zymography after 1 and 3 days of batimastat treatment (drug range, 0.2-2.0 microM). Athymic nude mice were given daily intraperitoneal injections of batimastat (30 mg/kg body weight) after resection of MDA-MB-435 primary tumors grown in their mammary fat pads; the volumes of tumor regrowths and the numbers and volumes of lung metastases were calculated; neovascularization in the regrowths was assessed by immunohistochemical analysis with an antibody directed against CD31, an endothelial cell antigen. The effect of batimastat treatment on the accumulation of mRNAs encoding specific MMPs and the tissue inhibitor of metalloproteinases-2 (TIMP-2) in cultured cells, primary tumors, and tumor regrowths was measured by RNA dot blotting and hybridization with complementary probes. Linear regression analysis, Student's t tests, and chi-squared analysis were used to evaluate the data.
RESULTS
The viability of cultured MDA-MB-435 cells was not affected by treatment with batimastat; however, measured activities for the 72-kd and 92-kd collagenases released by these cells were reduced after batimastat treatment. Intraperitoneal injection of batimastat significantly inhibited the local-regional regrowth of resected MDA-MB-435 tumors in athymic nude mice (in comparison with control mice, P = .035), and it reduced the incidence (P < .05), number (P = .0001), and total volume (P = .0001) of lung metastases. Batimastat treatment did not affect cellular levels of MMP or TIMP-2 mRNAs.
CONCLUSION
Batimastat inhibits human breast cancer regrowth and metastasis in a nude mouse xenograft model. Potential mechanisms for batimastat's inhibitory activity do not include direct cell toxicity or alteration of MMP or TIMP mRNA levels.