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人组织激肽释放酶结合蛋白(Kallistatin,KAL)是一种分泌型蛋白,其通过与胞外受体结合来调控多条下游信号通路,但尚未有KAL的胞内活性研究以及胞内KAL是否具有与胞外相似生物学活性的报道。通过构建无信号肽的KAL腺相关病毒表达载体(rAAV-NSK)来探索KAL蛋白的胞内活性。转染rAAV-NSK后,所有受试细胞均有KAL胞内表达但不分泌,其中HUVEC的增殖、迁移和小管形成均受到抑制;NCI-H460、NCI-H446和A549受到不同程度抑制。这一细胞水平研究,不仅证实了KAL的胞内活性,也暗示着KAL将可能作为一种“平衡因子”参与多靶点调控,也为解释目前有关KAL在PI3K-Akt信号通路的调控矛盾提供了新的思路。
Kallistatin (KAL), a secreted protein that regulates multiple downstream signaling pathways by binding to extracellular receptors, has not yet been investigated for intracellular activity of KAL and whether intracellular KAL is With similar biological activity of extracellular coverage. The intracellular activity of KAL protein was explored by constructing the signal peptide-free KAL-related adeno-associated virus expression vector (rAAV-NSK). After transfected with rAAV-NSK, all the cells tested showed the intracellular expression of KAL, but no secretion. The proliferation, migration and tubule formation of HUVEC were inhibited. NCI-H460, NCI-H446 and A549 were inhibited to some extent. This cellular level study not only confirms the intracellular activity of KAL, but also suggests that KAL may be involved as a “balance factor” in multi-target regulation, as well as to explain the current regulation of KAL in the PI3K-Akt signaling pathway Contradictions provide a new way of thinking.