成纤维细胞生长因子受体样蛋白1在口腔鳞状细胞癌中的表达及其对细胞增殖和迁移能力的影响
Expression of fibroblast growth factor receptor like 1 protein in oral squamous cell carcinoma and its influence on tumor cell proliferation and migration
作者：郑琛, 石超吉, 杜琳娟, 江银华, 苏吉梅
Author：Zheng Chen, Shi Chaoji, Du Linjuan, Jiang Yinhua, Su Jimei
收稿日期：2020-02-28 年卷（期）页码：2020,38(5):558-558-565
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：成纤维细胞生长因子受体样蛋白1,口腔鳞状细胞癌,细胞迁移,上皮间充质转化,RNA干扰,
Key words：fibroblast growth factor receptor like 1,oral squamous cell carcinoma,cell migration,epithelial-mesenchymal transition,RNA interference,
基金项目：
中文摘要
目的 检测成纤维细胞生长因子受体样蛋白1（FGFRL1）在口腔鳞状细胞癌（OSCC）中的表达情况,并探究其在OSCC细胞增殖和迁移中的作用。方法 利用Western blot检测FGFRL1蛋白在OSCC组织、癌旁正常组织、OSCC细胞株及正常上皮细胞中的表达;通过FGFRL1小干扰RNA（siRNA）干扰HN4细胞,影响FGFRL1蛋白的表达,利用CCK-8及Ki67实验检测FGFRL1对肿瘤细胞增殖能力的影响;细胞划痕及Transwell实验检测FGFRL1对肿瘤细胞迁移能力的影响;利用Western blot检测其对上皮间充质转化（EMT）相关指标蛋白的影响。结果 FGFRL1蛋白在OSCC组织中的表达量高于癌旁正常组织（t=2.820,P=0.047 8）;FGFRL1蛋白在OSCC细胞系中的表达量高于在HOK细胞中的表达量。实时荧光定量聚合酶链反应（qRT-PCR）结果显示,FGFRL1 RNA在HOK细胞中的表达量低于在OSCC细胞系中的表达量。将FGFRL1 siRNA转染的HN4细胞作为实验组,NC siRNA处理的HN4细胞作为对照组。Ki67免疫荧光试验结果显示,实验组与对照组在48 h（P=0.478 1）及72 h（P=0.334 2）细胞增殖活力差异无统计学意义。细胞划痕实验结果显示,在12 h（P=0.022 8）、24 h（P=0.005 1）及36 h（P=0.009 5）实验组细胞划痕面积百分比均比对照组小。Transwell实验结果显示,实验组在16 h（P=0.008 7）及24 h（P=0.008 6）细胞迁移数较对照组少。FGFRL1 siRNA干扰使得HN4细胞神经性钙黏附素蛋白和波形蛋白的表达量下降,上皮钙黏附素蛋白的表达量上升。结论 FGFRL1蛋白在OSCC组织中的表达量高于癌旁正常组织,在OSCC细胞株中的表达量高于正常上皮细胞。FGFRL1基因沉默对肿瘤细胞增殖无影响,但对肿瘤细胞EMT和细胞迁移具有一定的抑制作用。
英文摘要
ObjectiveThis study aims to investigate the expression of fibroblast growth factor receptor like 1 (FGFRL1) in oral squamous cell carcinoma (OSCC) and reveals its association with tumor cell proliferation and migration.MethodsWestern blot was performed to detect the expression of FGFRL1 protein in OSCC tissues, adjacent normal tissues, OSCC cell lines and normal epithelial cells. After knocking down of FGFRL1 in HN4 cells, CCK-8 and Ki67 assays were performed to detect cell proliferation, wounding healing assay and transwell were performed to detect cell-migration. Western blot was used to detect the expression of protein related to epithelial-mesenchymal transition (EMT).ResultsThe expression of FGFRL1 in OSCC tissues was higher than that in adjacent nontumor tissues, respectively (t=2.820, P=0.047 8). Moreover, the expression of FGFRL1 in OSCC cells was higher than that in HOK cells. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that FGFRL1 expression of FGFRL1 RNA in HOK cells was lower than that in OSCC cells. HN4 cells transfected with FGFRL1 siRNA were included in the experimental group, whereas HN4 cells treated with NC siRNA were included in the control group. CCK-8 experiment showed no significant difference between the experimental and control groups with regard to proliferation ability at 48 h (P=0.478 1) and 72 h (P=0.334 2). Migration experiment showed that the wound healing areas in the experimental group after 12 h (P=0.022 8), 24 h (P=0.005 1), and 36 h (P=0.009 5）were smaller than that in the control group. Transwell invasion assay showed that the number of invaded cells in the experimental group after 16 h (P=0.008 7) and 24 h (P=0.008 6) were lower than that in the control group. Knocking-down FGFRL1 up-regulated the expression of E-cadherin and down-regulated the expression of N-cadherin and Vimentin in HN4 cells.ConclusionFGFRL1 expression in the OSCC tissues was significantly higher than that in the adjacent nontumor tissues. FGFRL1 expression in the OSCC cells was significantly higher than that in the HOK cells, and FGFRL1 had no effect on cell proliferation but promoted tumor cell migration and EMT.
【关闭】
