The anoctamin (ANO) family consists of transmembrane proteins in 10 isoforms, and ANOs are broadly expressed in epithelial and non-epithelial tissues. Few studies have reported the function and activation mechanism of ANO9 in esophageal squamous cell carcinoma (ESCC), and the clinical significance of its expression remains unclear. The aims of the present study were to investigate the role of ANO9 in the regulation of tumor progression and its clinicopathological significance in ESCC. Methods In human ESCC cell lines KYSE150 and KYSE790, knockdown experiments were performed using ANO9 siRNA, and the effects on cell proliferation, cell cycle, apoptosis, invasion and migration were analyzed. The gene expression profiles of cells were examined using a microarray analysis. Immunohistochemical (IHC) analysis was performed on 57 primary tumor samples obtained from ESCC patients who underwent curative esophagectomy between 1999 and 2009 in Kyoto Prefectural University of Medicine. Results In an in vitro study, the depletion of ANO9 reduced cell proliferation, invasion and migration in KYSE150 and KYSE 790 cells. And, the depletion of ANO9 increased the number of cells in G0/G1 arrest and induced apoptosis in these cells. The results of the microarray analysis indicated that various centrosome-related genes such as CEP120, CNTRL and SPAST, were up- or down-regulated in ANO9-depleted KYSE150 cells. The IHC results showed that high expression of ANO9 was independent prognostic factor in ESCC patients (p = 0.025). Conclusion ANO9 played the important role in the cell cycle and progression of ESCC cells through the expression of centrosome-related genes. In addition, the high expression of ANO9 was a poor prognostic factor in ESCC patients. The results of the present study indicate that ANO9 has potential as a biomarker for cancer growth and as a therapeutic target for ESCC such as inhibitor or RNA interference of ANO9.

中文翻译：

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156 ANO9在人食管鳞状细胞癌中的表达和作用

anoctamin (ANO) 家族由 10 种同种型的跨膜蛋白组成，ANOs 在上皮和非上皮组织中广泛表达。鲜有研究报道ANO9在食管鳞状细胞癌（ESCC）中的功能和激活机制，其表达的临床意义尚不清楚。本研究的目的是探讨 ANO9 在调节肿瘤进展中的作用及其在 ESCC 中的临床病理学意义。方法在人ESCC细胞株KYSE150和KYSE790中，使用ANO9 siRNA进行敲低实验，分析其对细胞增殖、细胞周期、凋亡、侵袭和迁移的影响。使用微阵列分析检查细胞的基因表达谱。对 1999 年至 2009 年间在京都府立医科大学接受根治性食管切除术的 ESCC 患者获得的 57 例原发性肿瘤样本进行了免疫组织化学 (IHC) 分析。结果在一项体外研究中，ANO9 的消耗降低了 KYSE150 和 KYSE 790 细胞中的细胞增殖、侵袭和迁移。并且，ANO9 的消耗增加了 G0/G1 期阻滞的细胞数量并诱导这些细胞凋亡。微阵列分析的结果表明，各种中心体相关基因，如 CEP120、CNTRL 和 SPAST，在 ANO9 耗尽的 KYSE150 细胞中上调或下调。IHC 结果显示，ANO9 的高表达是 ESCC 患者的独立预后因素（p = 0.025）。结论 ANO9通过表达中心体相关基因在ESCC细胞的细胞周期和进程中发挥重要作用。此外，ANO9 的高表达是 ESCC 患者的不良预后因素。本研究的结果表明，ANO9 具有作为癌症生长的生物标志物和作为 ESCC 的治疗靶标（如 ANO9 的抑制剂或 RNA 干扰）的潜力。