TGF-β1对口腔癌相关成纤维细胞在二维和三维共培养条件下迁移的影响

Effects of TGF-β1 on the migration of oral cancer-associated fibroblasts in two and three dimensional co-cul-ture models

目的通过二维和三维条件下的细胞共培养模型观察转化生长因子-β1(transforming growth factor-β1,TGF-β1)对口腔癌相关成纤维细胞(carcinoma associated fibroblasts,CAFs)迁移的影响。方法二维培养条件下,以培养基中加入10 ng/mL TGF-β1刺激的CAFs为实验组,以未经处理的CAFs细胞为对照组,通过划痕实验、Transwell实验观察CAFs的迁移现象;通过逆转录病毒转染,培养绿色荧光蛋白(green fluorescent protein,GFP)阳性的CAFs细胞,建立人舌鳞癌细胞SCC25、GFP(+) CAFs和CAFs三维细胞共培养模型,以培养基中加入10 ng/mL TGF-β1刺激下培养的三维模型为实验组,以未加的TGF-β1的三维模型为对照组,观察在三维模型的细胞迁移现象。结果在二维培养条件下验证了10 ng/mL TGF-β1可促进口腔CAFs的迁移;成功建立SCC25、GFP(+) CAFs和CAFs三维细胞共培养模型,观察到CAFs和口腔癌细胞的迁移现象,但10 ng/mL的TGF-β1对上述细胞的迁移在三维条件下无明显促进作用。结论体外TGF-β1对口腔CAFs迁移的影响与二维和三维不同培养模式有关。

Objective To observe the effect of transforming growth factor-β1(TGF-β1)on the migration of oral car-cinoma associated fibroblasts(CAFs)with two-dimensional culture model and three-dimensional model.Methods Under two-dimensional culture conditions,CAFs stimulated by TGF-β1 with the addition of 10 ng/mL medium were used as the experimental group,and untreated CAFs were used as the control group.The migration of CAFs with the stimulation of TGF-β1 was measured by cell scratch assay and transwell assay.CAFs positive for green fluorescent pro-tein(GFP)were cultured by retrovirus transfection.Human tongue squamous cell carcinoma cells SCC25,GFP(+)CAFs and CAFs with three-dimensional cell co-culture models were established.The three-dimensional model cultured under the stimulation of TGF-β1 with 10 ng/mL medium was used as the experimental group,and the three-dimensional model without TGF-β1 was used as the control group.The migration of CAFs with the stimulation of TGF-β1 was also mea-sured by the three-dimensional models.Results It was verified that 10 ng/mL TGF-β1 promoted the migration of CAFs in the two-dimensional culture model.The three-dimensional co-culture models of SCC25,GFP(+)CAFs and CAFs were successfully established.The migration of SCC25 and CAFs was detected in the three-dimensional model.However,10 ng/mL TGF-β1 had little effect on their migration.Conclusion The effect of TGF-β1 in vitro on the migration of oral CAFs was associated with different culture models in two and three dimensions.