Role of the Na+/K+/2Cl- cotransporter NKCC1 in cell cycle progression in human esophageal squamous cell carcinoma
Role of the Na+/K+/2Cl- cotransporter NKCC1 in cell cycle progression in human esophageal squamous cell carcinoma
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DOI：
10.3748/wjg.v20.i22.6844
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发表时间：
2014-06-14
影响因子：
4.3
通讯作者：
Otsuji, Eigo
中科院分区：
文献类型：
--
作者：
Shiozaki, Atsushi;Nako, Yoshito;Otsuji, Eigo
目的：探讨Na+/K+/2Cl(-)协同转运蛋白1（NKCC1）在食管鳞状细胞癌（ESCC）细胞周期进程相关基因调控中的作用及其表达的临床病理意义。方法：对68例食管鳞状细胞癌患者的原发肿瘤样本进行免疫组织化学分析。通过蛋白质印迹分析人 ESCC 细胞系中的 NKCC1 表达。使用NKCC1小干扰RNA进行敲低实验，并分析对细胞周期进程的影响。通过微阵列分析细胞基因表达谱。结果：免疫组化染色显示NKCC1主要存在于癌细胞胞浆中，其表达与SCC的组织学分化程度相关。 NKCC1 在 KYSE170 细胞中高表达。这些细胞中 NKCC1 的耗竭通过 G(2)/M 期停滞抑制了细胞增殖。微阵列分析鉴定出 NKCC1 缺失的 KYSE170 中表达水平发生改变的 2527 个基因。通路分析显示，排名第一的经典通路是G(2)/M DNA损伤检查点调控通路，涉及MAD2L1、DTL、BLM、CDC20、BRCA1和E2F5。结论：ESCC中NKCC1的表达可能影响G(2)/M检查点，并可能与SCC的组织学分化程度有关。我们对 NKCC1 作为 ESCC 介质和/或生物标志物的作用有了更深入的了解。 (C) 2014 百事登出版集团有限公司版权所有。
AIM: To investigate the role of Na+/K+/2Cl(-) cotransporter 1 (NKCC1) in the regulation of genes involved in cell cycle progression and the clinicopathological significance of its expression in esophageal squamous cell carcinoma (ESCC).METHODS: An immunohistochemical analysis was performed on 68 primary tumor samples obtained from ESCC patients that underwent esophagectomy. NKCC1 expression in human ESCC cell lines was analyzed by Western blotting. Knockdown experiments were conducted using NKCC1 small interfering RNA, and the effects on cell cycle progression were analyzed. The gene expression profiles of cells were analyzed by microarray analysis.RESULTS: Immunohistochemical staining showed that NKCC1 was primarily found in the cytoplasm of carcinoma cells and that its expression was related to the histological degree of differentiation of SCC. NKCC1 was highly expressed in KYSE170 cells. Depletion of NKCC1 in these cells inhibited cell proliferation via G(2)/M phase arrest. Microarray analysis identified 2527 genes with altered expression levels in NKCC1depleted KYSE170. Pathway analysis showed that the top-ranked canonical pathway was the G(2)/M DNA damage checkpoint regulation pathway, which involves MAD2L1, DTL, BLM, CDC20, BRCA1, and E2F5.CONCLUSION: These results suggest that the expression of NKCC1 in ESCC may affect the G(2)/M checkpoint and may be related to the degree of histological differentiation of SCCs. We have provided a deeper understanding of the role of NKCC1 as a mediator and/or a biomarker in ESCC. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.