Department of Surgery, Division of Cardiothoracic Surgery, University of Colorado Denver School of Medicine, Aurora, CO, USA.
J Thorac Cardiovasc Surg. 2010 Mar;139(3):591-9; discussion 599. doi: 10.1016/j.jtcvs.2009.10.061.
Esophageal adenocarcinoma is thought to arise from lesions produced by chronic esophageal inflammation. Secretory phospholipase A(2) is an important mediator of mucosal response to gastroesophageal reflux, but its role in the function of mature cancer cells is unclear. We sought to determine the influence of group IIa secretory phospholipase A(2) on proliferation of human esophageal adenocarcinoma cells.
FLO-1 and OE33 cells derived from human esophageal adenocarcinoma were cultured with standard techniques. Cells were treated with 1-, 5-, 10-, and 20-mumol/L doses of 5-(4-benzyloxyphenyl)-4S-(7-phenylheptanoylamino)pentanoic acid, a specific inhibitor of group IIa secretory phospholipase A(2), for 72 hours. Gene for group IIa secretory phospholipase A(2)(PLA2G2A) was overexpressed and silenced with lentiviral infection techniques. Cell proliferation and viability were measured with standard 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and bromodeoxyuridine incorporation assays. All assays were performed in triplicate. PLA2G2A expression was measured with quantitative reverse transcriptase polymerase chain reaction; protein levels were detected with immunofluorescence microscopy. Statistical analysis was by analysis of variance with Fisher post hoc analysis.
Secretory phospholipase A(2) protein was found in both malignant esophageal adenocarcinoma cell lines. Treatment with specific group IIa secretory phospholipase A(2) inhibitor resulted in dose-dependent reductions in growth and cell number in both cell lines. Overexpression of PLA2G2A resulted in enhanced cancer cell growth, whereas gene knockdown attenuated growth.
Group IIa secretory phospholipase A(2) appears significant in growth and proliferation of human esophageal adenocarcinoma cells. Secretory phospholipase A(2) inhibition should be studied further regarding potential chemopreventive and therapeutic properties in esophageal adenocarcinoma.
食管腺癌被认为起源于慢性食管炎引起的病变。分泌型磷脂酶 A2（secretory phospholipase A2，sPLA2）是胃食管反流对黏膜的重要介质反应，但它在成熟癌细胞功能中的作用尚不清楚。我们试图确定 IIa 组分泌型磷脂酶 A2（group IIa secretory phospholipase A2，PLA2G2A）对人食管腺癌细胞增殖的影响。
采用标准技术培养源自人食管腺癌的 FLO-1 和 OE33 细胞。用 1、5、10 和 20μmol/L 剂量的 5-（4-苄氧基苯基）-4S-（7-苯庚酰氨基）戊酸处理细胞 72 小时，这是一种 IIa 组分泌型磷脂酶 A2 的特异性抑制剂。通过慢病毒感染技术过表达和沉默基因 PLA2G2A。用标准的 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐和溴脱氧尿苷掺入测定法测量细胞增殖和活力。所有测定均一式三份进行。用定量逆转录聚合酶链反应（quantitative reverse transcriptase polymerase chain reaction，qRT-PCR）测量 PLA2G2A 表达，用免疫荧光显微镜检测蛋白水平。统计分析采用方差分析和 Fisher 事后分析。
在两种恶性食管腺癌细胞系中均发现分泌型磷脂酶 A2 蛋白。用特异性 IIa 组分泌型磷脂酶 A2 抑制剂处理可导致两种细胞系的生长和细胞数量呈剂量依赖性减少。PLA2G2A 的过表达导致癌细胞生长增强，而基因敲低则减弱了生长。
IIa 组分泌型磷脂酶 A2 在人食管腺癌细胞的生长和增殖中具有重要作用。进一步研究分泌型磷脂酶 A2 抑制在食管腺癌中的潜在化学预防和治疗特性。
