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。

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。

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_{2 d}= 17.941，P_{2 d}= 0.003；t_{3 d}= 24.430，P_{3 d}= 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_{2 d}= 50.230，P_{2 d}= 0.005；t_{3 d}= 26.883，P_{3 d}= 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_{2 d}= 0.026；t_{3 d}= 6.423，P_{3 d}= 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（t_{2 d}= 20.693，P_{2 d}= 0.072；t_{3 d}= 9.875，P_{3 d}= 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_{2 d}= 21.829，P_{2 d}= 0.020；t_{3 d}= 9.853，P_{3 d}= 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_{2 d}= 30.693，P_{2 d}= 0.015；t_{3 d}= 21.173，P_{3 d}= 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_{2 d}= 7.187，P_{2 d}= 0.019；t_{4 d}= 6.631，P_{4 d}= 0.022；t_{6 d}= 6.690，P_{6 d}= 0.022），间质标记蛋白Vimentin（1.089 ± 0.134、0.706 ± 0.025、0.620 ± 0.010）表达在2、4、6 d处与对照组相比表达升高（t_{2 d}= 6.948，P_{2 d}= 0.020；t_{4 d}= 16.710，P_{4 d}= 0.004；t_{6 d}= 6.157，P_{6 d}= 0.025），EMT转录因子snail在2 d时（1.14 ± 0.17）表达与对照组（0.77 ± 0.10）相比表达升高（t= 3.794，P= 0.015），EMT转录因子slug在2 d时（1.85 ± 0.11）表达与对照组（0.93 ± 0.02）相比表达升高（t= 15.385，P= 0.014）；与对照组（20.0 ± 2.0）相比，TGF-β1处理后细胞迁移能力（45.7 ± 11.6）显著增加（t= 4.529，P= 0.017），细胞侵袭能力（58.7 ± 5.0）较对照组（22.3 ± 1.5）明显升高（t= 15.571，P= 0.015）。

TGF-β1增强糖酵解，并促进TSCC细胞EMT及迁移和侵袭。

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) .

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.

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 (t_{2 d}= 17.941, P_{2 d}= 0.003; t_{3 d}= 24.430, P_{3 d}= 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 (t_{2 d}= 50.230, P_{2 d}= 0.005; t_{3 d}= 26.883, P_{3 d}= 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 (t_{2 d}= 6.111, P_{2 d}= 0.026; t_{3 d}= 6.423, P_{3 d}= 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 (t_{2 d}= 20.693, P_{2 d}= 0.072; t_{3 d}= 9.875, P_{3 d}= 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_{2 d}= 21.829, P_{2 d}= 0.020; t_{3 d}= 9.853, P_{3 d}= 0.022) ; The expression level of GLUT1 (0.503 ± 0.007, 0.589 ± 0.019) in TGF-β1 treatment group was higher than control group, and both showed statistically significant difference (t_{2 d}= 30.693, P_{2 d}= 0.015; t_{3 d}= 21.173, P_{3 d}= 0.012) . Under the TGF-β1 induction, the morphology of TSCC SCC9 cells changed from pebble to long spindle shape compared with the control group; The expression level of E-cadherin (0.69 ± 0.03, 0.67 ± 0.04, 0.65 ± 0.04) in TGF-β1 treatment group was lower than control group, and both showed statistically significant difference (t_{2 d}= 7.187, P_{2 d}= 0.019; t_{4 d}= 6.631, P_{4 d}= 0.022; t_{6 d}= 6.690, P_{6 d}= 0.022) ; The expression level of Vimentin (1.089 ± 0.134, 0.706 ± 0.025, 0.620 ± 0.010) in TGF-β1 treatment group was higher than control group, and both showed statistically significant difference (t_{2 d}= 6.948, P_{2 d}= 0.020; t_{4 d}= 16.710, P_{4 d}= 0.004; t_{6 d}= 6.157, P_{6 d}= 0.025) ; The expression level of snail (1.14 ± 0.17) in TGF-β1 treatment group was higher than control group (0.77 ± 0.10) , and both showed statistically significant difference (t= 3.794, P= 0.014) ; The expression level of slug (1.85 ± 0.11) in TGF-β1 treatment group was higher than control group (0.93 ± 0.02) , and both showed statistically significant difference (t= 15.385, P= 0.015) . The cell migration ability of SCC9 cell (45.7 ± 11.6) was increased under TGF-β1 treatment compared with control group (20.0 ± 2.0; t= 4.529, P= 0.017) ; The cell invasion ability of SCC9 cell (58.7 ± 5.0) was increased under TGF-β1 treatment compared with control group (22.3 ± 1.5; t= 15.571, P= 0.015) .

TGF-β1 enhances glycolysis and promotes EMT, cell migration and invasion of tongue squamous cell carcinoma.