Rho Kinase ROCK2 Mediates Acid-Induced NADPH Oxidase NOX5-S Expression in Human Esophageal Adenocarcinoma Cells.
Rho Kinase ROCK2 Mediates Acid-Induced NADPH Oxidase NOX5-S Expression in Human Esophageal Adenocarcinoma Cells.
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DOI：
10.1371/journal.pone.0149735
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Cao W
中科院分区：
文献类型：
--
作者：
Hong J;Li D;Cao W
从Barrett食管（BE）发展为食管腺癌（EA）的机制尚未完全了解。我们已经证明，NOX 5-S可能参与了这一进程。然而，酸如何上调NOX 5-S尚不清楚。我们发现，在BE粘膜活检和FLO-1 EA细胞中，Rho激酶（ROCK）抑制剂Y27632显著降低了酸诱导的NOX 5-S表达增加。此外，酸处理显着增加FLO-1细胞中的Rho激酶活性。敲低Rho激酶ROCK 2可显著降低酸诱导的NOX 5-S表达和H2 O2产生的增加，但敲低ROCK 1则无此作用。相反，组成型活性ROCK 2的过表达，而不是组成型活性ROCK 1，显着增强NOX 5-S的表达和H2 O2的生产。此外，酸诱导的Rho激酶活性和NOX 5-S mRNA表达的增加被FLO-1和OE 33细胞中钙的去除所阻断。钙离子载体A23187显著增加Rho激酶活性和NOX 5-S mRNA表达。我们的结论是，酸诱导的NOX 5-S的表达和H2 O2的生产增加可能取决于ROCK 2的激活，而不是ROCK 1，在EA细胞。酸诱导的Rho激酶的激活可能是通过细胞内钙的增加来介导的。BE患者中存在的持续酸反流可能增加细胞内钙，激活ROCK 2，从而上调NOX 5-S。高水平的活性氧来源于NOX 5-S可能会导致DNA损伤，从而有助于从BE到EA的进展。
Mechanisms of the progression from Barrett’s esophagus (BE) to esophageal adenocarcinoma (EA) are not fully understood. We have shown that NOX5-S may be involved in this progression. However, how acid upregulates NOX5-S is not well known. We found that acid-induced increase in NOX5-S expression was significantly decreased by the Rho kinase (ROCK) inhibitor Y27632 in BE mucosal biopsies and FLO-1 EA cells. In addition, acid treatment significantly increased the Rho kinase activity in FLO-1 cells. The acid-induced increase in NOX5-S expression and H2O2 production was significantly decreased by knockdown of Rho kinase ROCK2, but not by knockdown of ROCK1. Conversely, the overexpression of the constitutively active ROCK2, but not the constitutively active ROCK1, significantly enhanced the NOX5-S expression and H2O2 production. Moreover, the acid-induced increase in Rho kinase activity and in NOX5-S mRNA expression was blocked by the removal of calcium in both FLO-1 and OE33 cells. The calcium ionophore A23187 significantly increased the Rho kinase activity and NOX5-S mRNA expression. We conclude that acid-induced increase in NOX5-S expression and H2O2 production may depend on the activation of ROCK2, but not ROCK1, in EA cells. The acid-induced activation of Rho kinase may be mediated by the intracellular calcium increase. It is possible that persistent acid reflux present in BE patients may increase the intracellular calcium, activate ROCK2 and thereby upregulate NOX5-S. High levels of reactive oxygen species derived from NOX5-S may cause DNA damage and thereby contribute to the progression from BE to EA.