转化生长因子β1增强糖酵解促进舌鳞状细胞癌细胞上皮间充质转化及迁移与侵袭  

Transforming growth factor-β1 enhances glycolysis and promotes epithelial mesenchymal transition,cell migration and invasion of tongue squamous cell carcinoma

作　　者：李文清[1] 刘海潮[1] 梁建锋[1] 陈冠辉[1] 竺越[1] 张鸣 侯劲松[1] Li Wenqing;LiuHaichao;Liang Jianfeng;Chen Guanhui;Zhu Yue;Zhang Ming;Hou Jinsong(Guanghua School ofStomatology,Hospital of Stomatology,Sun Yat ？ sen University,Guangdong Provincial Key Laboratory ofStomatology,Guangzhou 510055,China)

机构地区：[1]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2018年第3期144-151,共8页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(81572660);广东省自然科学基金(2015A030313034);中山大学临床医学研究5010计划(2015018)

摘　　要：目的研究转化生长因子β1(TGF-β1)对舌鳞状细胞癌(TSCC)糖酵解活性和上皮间充质转化(EMT)及迁移与侵袭的影响。方法利用10 ng/mL TGF-β1处理TSCC SCC9细胞1、2、3 d,收集细胞上清液检测葡萄糖和乳酸表达变化;蛋白免疫印迹法(Western blot)检测TGF-β1处理1、2、3 d后糖酵解关键酶表达变化;应用10 ng/mL TGF-β1处理TSCC SCC9细胞2、4、6 d,在倒置显微镜下定时观察细胞形态;Western blot检测TGF-β1诱导2、4、6 d后EMT上皮标记蛋白E-cadherin、间质标记蛋白Vimentin、Snail和Slug的表达变化;Transwell小室检测细胞迁移和侵袭能力;同等培养条件下未经TGF-β1处理的为对照组。SPSS 13.0统计软件进行数据分析。结果在TGF-β1诱导条件下,TSCC SCC9细胞葡萄糖摄取量在2和3 d时[(34.1±1.2)、(47.1±2.3)mmol]较对照组显著升高(t_(2d)=17.941,P2 d=0.003;t_(3d)=24.430,P_(3d)=0.002),同时SCC9细胞乳酸生成量在2和3 d时[(46.4±1.0)、(60.2±2.0)mmol]较对照组明显增加(t_(2d)=50.230,P_(2d)=0.005;t_(3d)=26.883,P_(3d)=0.004);TGF-β1处理后,糖酵解关键酶HK2表达在2和3 d时(1.21±0.04、1.30±0.06)均高于对照组,差异有统计学意义(t_(2d)=6.111,P_(2d)=0.026;t_(3d)=6.423,P_(3d)=0.023);糖酵解关键酶PKM2表达在2和3 d时(1.048±0.002、1.071±0.010)与对照组相比,差异无统计学意义(t2 d=20.693,P2 d=0.072;t_(3d)=9.875,P_(3d)=0.081);糖酵解关键酶PFKP在2和3 d时(0.820±0.010、0.839±0.036)表达较对照组明显升高(t_(2d)=21.829,P_(2d)=0.020;t_(3d)=9.853,P_(3d)=0.022);糖酵解关键酶GLUT1表达在2和3 d时(0.503±0.007、0.589±0.019)均高于对照,差异具有统计学意义(t_(2d)=30.693,P_(2d)=0.015;t_(3d)=21.173,P_(3d)=0.012)。在TGF-β1诱导下,与对照组相比,TSCC SCC9细胞从鹅卵石状变为长梭形,同时EMT上皮标记蛋白E-cadherin表达在2、4和6 d时(0.69±0.03、0.67±0.04、0.65±0.04)较对照组降低,差异有统计学意义(t_(2d)=7.187,P_(2d)=0.019;t_(4 d)=6.631,P_(4 d)=0.022;t_(6 d)=6.690,Objective To investigate the influence of transforming growth factor-β1（TGF-β1）on glycolysis,inducing epithelial mesenchymal transition（EMT）,cell migration and invasion of tongue squamous cell carcinoma（TSCC）. Methods TSCC SCC9 cells were treated with 10 ng/mL TGF-β1 for 1,2 and 3 days and the cell supernatant was collected to detect changes in glucose and lactate. Western blot was used to detect the expression of glycolysis key enzymes following 1,2,3 days of treatment of TGF-β1.TSCC SCC9 cells were treated with 10 ng/mL TGF-β1 for 2,4,6 days,and the morphology of the cells was observed under inverted microscope. Western blot was used to detect the expression of E-cadherin,Vimentin,Snail and Slug following 2,4,6 days of treatment of TGF-β1. Transwell chamber was performed to detect cell migration and invasion. Statistical analysis was performed with SPSS 13.0 software.Results Under the TGF-β1 induction,the glucose uptake（34.1 ± 1.2,47.1 ± 2.3）of SCC9 cells was significantly higher than that in the control group,and both showed statistically significant difference（t2d =17.941,P2d = 0.003;t3d = 24.430,P3d = 0.002）. The lactic acid production（46.4 ± 1.0,60.2 ± 2.0）in SCC9 cells was significantly higher than that in the control group,and both showed statistically significant difference（t2d = 50.230,P2d = 0.005;t3d = 26.883,P3d = 0.004）. The expression level of HK2（1.21 ± 0.04,1.30 ± 0.06）in TGF-β1 treatment group was higher than control group,and both showed statistically significant difference（t2d = 6.111,P2d = 0.026;t3d = 6.423,P3d = 0.023）;the expression level of PKM2（1.048±0.002,1.071±0.010）in TGF-β1 treatment group showed no significant difference compared with the control group,and both showed no statistically significant difference（t2d = 20.693,P2d = 0.072;t3d =9.875,P3d = 0.081）;the expression level of PFKP（0.820±0.010,0.839±0.036）in TGF-β1 treatment group was higher than control group,and both showed statistically significant difference（t

关 键 词：舌 癌 鳞状细胞 转化生长因子Β1 糖酵解 上皮细胞-间充质转换 细胞迁移分析 肿瘤侵袭 

分 类 号：R739.86[医药卫生—肿瘤]