Zhang Yanli, Wang Qiuyan, Ma Ailing, Li Yunhong, Li Rui, Wang Yin
Basic Medical College, Center of Scientific Technology, Ningxia Medical University, Ningxia Key Laboratory of Cerebrocranial Diseases, Yinchuan, Ningxia 750004, P.R. China.
The General Hospital of Ningxia Medical University, Yinchuan, Ningxia 750004, P.R. China.
Oncol Rep. 2014 May;31(5):2298-304. doi: 10.3892/or.2014.3095. Epub 2014 Mar 19.
The Toll-like receptor 9 (TLR9) plays a crucial role in both innate and adaptive immune responses against infection and danger signals. Stimulation of TLR9 has been linked to invasion in various cancer cells in vitro. The present study evaluated the expression of TLR9 in human esophageal cancer (EC) cells and normal and malignant esophageal squamous epithelium, and examined the association between TLR9 expression, clinicopathological variables, and EC patient outcome. We further characterized the direct effects of TLR9 agonist CpG oligonucleotides (CpG ODN) and inhibitor chloroquine (CQ), on the proliferation and invasion of EC cells in vitro. RT-PCR, western blot, flow cytometry and immunohistochemical analysis were used to determine the expression of TLR9 in EC cell line TE10, and 90 cases of esophageal squamous cell carcinoma, including 30 cases of adjacent esophageal epithelium. The TLR9 expression was compared with tumor size, location, grade, stage and proliferation. We found basal expression of TLR9 in TE10 cells. Esophageal carcinomas exhibited TLR9 expression that was positively associated with tumor size, location and TNM stage (P<0.05). CpG ODN significantly enhanced the invasion of TE10 cells, which could be abrogated by a TLR9 inhibitor CQ. CpG ODN led to activation of NF‑κB and enhanced expression of matrix metalloproteinase (MMP)-2, MMP-7 and cyclooxygenase-2 (COX-2) mRNA. Expression of TLR9 in EC suggests a role of TLR9 related to cell proliferation and differentiation. Our findings indicate that TLR9 may represent a novel therapeutic target in this disease.
Toll样受体9（TLR9）在针对感染和危险信号的先天性和适应性免疫反应中均发挥着关键作用。TLR9的刺激已与体外各种癌细胞的侵袭相关联。本研究评估了TLR9在人食管癌（EC）细胞以及正常和恶性食管鳞状上皮中的表达，并研究了TLR9表达、临床病理变量与EC患者预后之间的关联。我们进一步表征了TLR9激动剂CpG寡核苷酸（CpG ODN）和抑制剂氯喹（CQ）对EC细胞体外增殖和侵袭的直接影响。采用逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法、流式细胞术和免疫组织化学分析来确定TLR9在EC细胞系TE10以及90例食管鳞状细胞癌（包括30例相邻食管上皮）中的表达。将TLR9表达与肿瘤大小、位置、分级、分期和增殖情况进行比较。我们发现TE10细胞中有TLR9的基础表达。食管癌中TLR9的表达与肿瘤大小、位置和TNM分期呈正相关（P<0.05）。CpG ODN显著增强了TE10细胞的侵袭能力，而这种作用可被TLR9抑制剂CQ消除。CpG ODN导致核因子κB（NF-κB）激活，并增强了基质金属蛋白酶（MMP）-2、MMP-7和环氧化酶-2（COX-2）mRNA的表达。TLR9在EC中的表达表明其在细胞增殖和分化方面发挥作用。我们的研究结果表明，TLR9可能是这种疾病的一个新的治疗靶点。
