Department of Gastroenterological Surgery (Surgery II), Nagoya University Graduate School of Medicine, Nagoya, Japan.
Department of Innovative Research and Education for Clinicians and Trainees (DiRECT), Fukushima Medical University Hospital, Fukushima City, Fukushima, Japan.
Ann Surg Oncol. 2020 Jul;27(7):2559-2568. doi: 10.1245/s10434-019-08189-8. Epub 2020 Feb 12.
We aimed to clarify the role of potassium voltage-gated channel subfamily J member 15 (KCNJ15) in esophageal squamous cell carcinoma (ESCC) cells and its potential as a prognosticator in ESCC patients.
KCNJ15 transcription levels were evaluated in 13 ESCC cell lines and polymerase chain reaction (PCR) array analysis was conducted to detect coordinately expressed genes with KCNJ15. The biological functions of KCNJ15 in cell invasion, proliferation, migration, and adhesion were validated through small interfering RNA-mediated knockdown experiments. Cell proliferation was further evaluated through the forced expression experiment. KCNJ15 expression was detected in 200 ESCC tissues by quantitative real-time reverse transcription PCR (qRT-PCR) and analyzed in 64 representative tissues by immunohistochemistry. Correlations between KCNJ15 expression levels and clinicopathological features were also analyzed.
The KCNJ15 expression levels varied widely in ESCC cell lines and correlated with COL3A1, JAG1, and F11R. Knockdown of KCNJ15 expression significantly repressed cell invasion, proliferation, and migration of ESCC cells in vitro. Furthermore, overexpression of KCNJ15 resulted in increased cell proliferation. Patients were stratified using the cut-off value of KCNJ15 messenger RNA (mRNA) levels in 200 ESCC tissues using receiver operating characteristic curve analysis; the high KCNJ15 expression group had significantly shorter overall and disease-free survival times. In multivariable analysis, high expression of KCNJ15 was identified as an independent poor prognostic factor. Staining intensity of in situ KCNJ15 protein expression tended to be associated with KCNJ15 mRNA expression levels.
KCNJ15 is involved in aggressive tumor phenotypes of ESCC cells and its tissue expression levels may be useful as a prognosticator of patients with ESCC.
本研究旨在阐明钾电压门控通道亚家族 J 成员 15(KCNJ15)在食管鳞状细胞癌(ESCC)细胞中的作用及其作为 ESCC 患者预后标志物的潜力。
通过聚合酶链反应(PCR)阵列分析评估 13 种 ESCC 细胞系中 KCNJ15 的转录水平,并检测与 KCNJ15 协调表达的基因。通过小干扰 RNA 介导的敲低实验验证 KCNJ15 在细胞侵袭、增殖、迁移和黏附中的生物学功能。通过强制表达实验进一步评估细胞增殖。通过实时定量逆转录 PCR(qRT-PCR)检测 200 例 ESCC 组织中的 KCNJ15 表达,并在 64 例代表性组织中通过免疫组织化学检测 KCNJ15 表达。还分析了 KCNJ15 表达水平与临床病理特征之间的相关性。
ESCC 细胞系中 KCNJ15 的表达水平差异很大,与 COL3A1、JAG1 和 F11R 相关。敲低 KCNJ15 表达显著抑制 ESCC 细胞的体外侵袭、增殖和迁移。此外,KCNJ15 的过表达导致细胞增殖增加。使用 200 例 ESCC 组织的 KCNJ15 信使 RNA(mRNA)水平的截断值对患者进行分层;高 KCNJ15 表达组的总生存期和无病生存期明显缩短。在多变量分析中,高表达 KCNJ15 被确定为独立的不良预后因素。原位 KCNJ15 蛋白表达的染色强度倾向于与 KCNJ15 mRNA 表达水平相关。
KCNJ15 参与 ESCC 细胞的侵袭性肿瘤表型,其组织表达水平可能作为 ESCC 患者的预后标志物。