SphK1在舌鳞癌中的表达及对Tca8113P160舌鳞癌细胞增殖、迁移及侵袭影响的体外研究

发布时间：2018-04-10 14:20

本文选题：舌鳞状细胞癌 + 鞘氨醇激酶1　；参考：《天津医科大学》2017年硕士论文

【摘要】：研究目的:检测鞘氨醇激酶1(sphingosine kinase 1,Sph K1)在舌鳞状细胞癌(tongue squamous cell carcinoma,TSCC)中的表达及其与临床病理特征和淋巴管生成之间的关系,并探索其在舌鳞状细胞癌发生发展中的作用及可能的分子机制。研究方法:1.收集天津市口腔医院50例舌鳞状细胞癌手术标本作为研究对象,应用免疫组化法,检测Sph K1蛋白的表达水平以及淋巴管密度(lymphatic vessel density,LVD),并分析Sph K1蛋白的表达水平与临床病理特征及淋巴管生成间的关系;2.体外实验使用Sph K1抑制剂N,N-二甲基鞘氨醇(N,N-dimethylsphingosine,DMS)干预Tca8113P160舌鳞状细胞癌细胞系,CCK-8法检测DMS对细胞增殖的影响,Transwell小室模型检测DMS对细胞迁移和侵袭能力的影响,Western blot检测DMS对Sph K1、FAK、p-FAK、E-cadherin和N-cadherin蛋白表达的影响;另外用FAK抑制剂PF573228干预Tca8113P160舌鳞状细胞癌细胞系,CCK-8法检测PF573228对细胞增殖的影响,Transwell小室模型检测PF573228对细胞迁移和侵袭能力的影响,Western blot检测PF573228对E-cadherin和N-cadherin蛋白表达的影响。研究结果:1.Sph K1在舌鳞状细胞癌组织中阳性表达率(62.0%)明显高于白斑(42.9%)、正常舌粘膜组织(0%),差异有统计学意义(P0.05)。Sph K1表达水平与患者的临床分期(P=0.002)、有无淋巴结转移(P=0.001)、T分期(P=0.019)及组织学分级(P=0.024)密切相关,与患者的性别(P=0.341)、年龄(P=0.786)、吸烟情况(P=0.547)及饮酒情况(P=0.566)无显著相关性(P0.05);淋巴结转移阳性组LVD(10.76±3.33)明显高于淋巴结转移阴性组LVD(7.84±2.53),差异有统计学意义(P0.05);Sph K1高表达组LVD(10.81±2.86)明显高于Sph K1低表达组LVD(6.84±2.32),差异有统计学意义(P0.05)。2.DMS和PF573228均能抑制Tca8113P160细胞的增殖,呈时间和剂量依赖性;DMS和PF573228均能抑制细胞的迁移和侵袭能力(P0.05);Western blot结果显示DMS抑制Sph K1蛋白的表达,同时下调FAK、p-FAK、N-cadherin蛋白的表达,上调E-cadherin蛋白的表达(P0.05),PF573228下调N-cadherin蛋白的表达,上调E-cadherin蛋白的表达(P0.05)。结论:1.Sph K1与舌鳞状细胞癌的发生发展及转移密切相关,可能通过促进肿瘤淋巴管生成影响舌鳞状细胞癌颈淋巴结转移。2.Sph K1通过FAK通路调控上皮间质转化(epithelial-mesenchymal transition,EMT)进而影响Tca8113P160舌鳞状细胞癌细胞的迁移和侵袭。
[Abstract]:Objective: to detect the expression of sphingosine kinase (1(sphingosine kinase 1) in tongue squamous cell carcinoma (TSCC) and its relationship with clinicopathological features and lymphangiogenesis.To explore its role in the development of squamous cell carcinoma of tongue and its possible molecular mechanism.Research method: 1.Fifty cases of squamous cell carcinoma of tongue were collected from Tianjin Stomatology Hospital.The expression level of Sph K1 protein and lymphatic vessel density were detected. The relationship between the expression level of Sph K1 protein and clinicopathological features and lymphangiogenesis was analyzed.To influence the expression of E-cadherin and N-cadherin protein in Sph K1FAKP p-FAKT.In addition, FAK inhibitor PF573228 was used to detect the effect of PF573228 on cell proliferation in Tca8113P160 squamous cell carcinoma cell line (Tca8113P160) by CCK-8 method. Transwell chamber model was used to detect the effect of PF573228 on cell migration and invasion. Western blot was used to detect the effect of PF573228 on the expression of E-cadherin and N-cadherin protein.P0. 019) and histologic grading P0. 024) were closely related.In K1 low expression group, LVD(6.84 卤2.32, the difference was statistically significant (P 0.05). 2. DMS and PF573228 could inhibit the proliferation of Tca8113P160 cells.In a time-and dose-dependent manner, both DMS and PF573228 could inhibit cell migration and invasion. The results of Western blot showed that DMS inhibited the expression of Sph K1 protein, down-regulated the expression of FAK-p-FAK-N-cadherin protein, up-regulated the expression of E-cadherin protein and down-regulated the expression of N-cadherin protein.The expression of E-cadherin protein was up-regulated.Conclusion: 1. SphK1 is closely related to the occurrence, development and metastasis of tongue squamous cell carcinoma.SphK1 may affect the cervical lymph node metastasis of tongue squamous cell carcinoma by promoting lymphangiogenesis. 2. SphK1 regulates epithelial mesenchymal transition via FAK pathway and then affects the migration and invasion of Tca8113P160 squamous cell carcinoma.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.86

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