Bafilomycin A1 prevents maturation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells
Bafilomycin A1 prevents maturation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells
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DOI：
10.1247/csf.23.33
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发表时间：
1998-02-01
影响因子：
1.5
通讯作者：
Tashiro, Y
中科院分区：
文献类型：
--
作者：
Yamamoto, A;Tagawa, Y;Tashiro, Y
我们研究了一种有效的、特异性的液泡H+ ATP酶（V-ATP酶）抑制剂巴弗洛霉素A（1）对大鼠肝癌细胞系H-4-II-E细胞自噬过程的影响。为了诱导自噬，将细胞从含有12%胎牛血清的Dulbecco改良Eagle培养基转移到Hanks平衡盐溶液中。当把巴弗洛霉素A（1）加入Hanks平衡盐溶液中时，内源性蛋白质降解被强烈抑制，大量自噬体在H-4-II-E细胞中积累，而自溶体数量减少。在存在巴弗洛霉素A的情况下，在自噬体中未检测到酸性磷酸酶活性（1），这表明自噬体和溶酶体之间的融合被这种药物干扰。融合的抑制是可逆的，去除抑制剂后，自噬体变为自溶酶体。巴弗洛霉素A（1）也阻止了自噬泡内吞HRP的出现。这些结果表明，V-ATP酶对自噬体或溶酶体内腔的酸化对于自噬体与溶酶体之间的融合至关重要。
We studied the effects of bafilomycin A(1), a potent and specific inhibitor of vacuolar H+ ATPase (V-ATPase), on the process of autophagy in rat hepatoma cell line, H-4-II-E cells. To induce autophagy, cells were transferred from Dulbecco's modified Eagle medium containing 12% fetal calf serum into Hanks' balanced salt solution. When bafilomycin A(1) was added to Hanks' balanced salt solution, endogenous protein degradation was strongly inhibited and numerous autophagosomes accumulated in H-4-II-E cells, whereas autolysosomes decreased in number. Acid phosphatase activity was not detected ire the autophagosomes which accumulated in the presence of bafilomycin A(1), suggesting that fusion between autophagosomes and lysosomes was disturbed by this drug. Inhibition of the fusion was reversible, and the autophagosomes changed into autolysosomes after the removal of the inhibitor. Bafilomycin A(1) also prevented the appearance of endocytosed HRP in autophagic vacuoles. These results suggested that acidification of the lumenal space of autophagosomes or lysosomes by V-ATPase is important for the fusion between autophagosomes and lysosomes.