THROMBIN MODULATES AND REVERSES NEURO-BLASTOMA NEURITE OUTGROWTH
THROMBIN MODULATES AND REVERSES NEURO-BLASTOMA NEURITE OUTGROWTH
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DOI：
10.1073/pnas.85.10.3440
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发表时间：
1988-05-01
影响因子：
11.1
通讯作者：
CUNNINGHAM, DD
中科院分区：
文献类型：
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作者：
GURWITZ, D;CUNNINGHAM, DD
先前的研究表明，当从含血清培养基转换为无血清培养基时，培养中的神经母细胞瘤细胞和几种类型的原代神经元细胞迅速延伸神经突。目前对克隆的神经母细胞瘤细胞的研究表明，凝血酶在2 nM时阻断了这种自发分化，半最大效力为50 pM。这需要凝血酶的催化活性，并在凝血酶去除后逆转。凝血酶也导致细胞在无血清培养基中撤退他们的神经突延伸，即使在100倍的浓度。凝血酶的特异性测定表明，在神经母细胞瘤培养物的含血清培养基中检测到的凝血酶来源于血清，并且可能是血清维持神经母细胞瘤细胞处于未分化状态的大部分已知能力的原因。这得到了以下发现的支持：肝素的加入降低了含血清培养基中的凝血酶浓度，并刺激了含血清培养基中神经母细胞瘤细胞的神经突生长。对凝血酶通过其他试剂调节神经突生长的能力的研究表明，它阻断并逆转了两种凝血酶抑制剂的神经突生长活性：蛋白酶nexin-1（与神经胶质源性神经突促进因子相同）和水蛭素。然而，凝血酶并不能阻断二丁酰cAMP或前列腺素E的促神经突活性。这些结果表明，凝血酶在控制神经突生长的具体作用。
Previous studies have shown that neuroblastoma cells and several types of primary neuronal cells in culture rapidly extend neurites when switched from serum-containing to serum-free medium. The present studies on cloned neuroblastoma cells show that thrombin blocked this spontaneous differentiation at 2 nM with a half-maximal potency of 50 pM. This required the catalytic activity of thrombin and reversed upon thrombin removal. Thrombin also caused cells in serum-free medium to retreat their neurite extension even at 100-fold higher concentrations. A specific assay for thrombin indicated that thrombin detected in serum-containing medium from neuroblastoma cultures was derived from serum and that it was likely responsible for much of the known capacity of serum to maintain neuroblastoma cells in a nondifferentiated state. This was supported by the finding that heparin addition reduced the thrombin concentration in serum-containing medium and stimulated neurite outgrowth from neuroblastoma cells in serum-containing medium. Studies on the ability of thrombin to modulate neurite outgrowth by other agents showed that it blocked and reversed the neurite outgrowth activity of two thrombin inhibitors: protease nexin-1 (which is identical to glial-derived neurite-promoting factor) and hirudin. Thrombin, however, did not block the neurite-promoting activity of dibutyryl cAMP or prostaglandin E. These results suggest a specific role for thrombin in control of neurite outgrowth.