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目的探讨转化生长因子β1(transforming growth factor-β1,TGF-β1)联合表皮生长因子(epithelial growth factor,EGF)诱导上皮向间质转化对人喉癌细胞株Hep-2侵袭能力的影响。方法应用TGF-β1单独刺激和TGF-β1联合EGF刺激人喉癌细胞株Hep-2后24、48 h观察细胞形态学的动态变化。应用Transwell检测细胞侵袭能力及RT-PCR和Western blot技术检测细胞上皮钙依赖黏附蛋白(E-cadherin)和波形蛋白(vimentin)的表达。结果①TGF-β1单独处理48 h组和TGF-β1联合EGF处理24、48 h组均能刺激喉癌细胞株Hep-2发生细胞形态改变。②TGF-β1联合EGF处理组无论是24 h还是48 h,其Hep-2细胞的侵袭能力都明显强于相同时段TGF-β1单独处理组(P<0.05);TGF-β1单独处理48 h组细胞侵袭能力又明显强于对照组(P<0.05)。③TGF-β1联合EGF处理组E-cadherin的表达降低,而Vimentin表达却明显上升。结论 EGF能够协同TGF-β1诱导上皮向间质转化,从而使喉癌具有更强的侵袭能力。
Objective To investigate the effect of transforming growth factor-β1 (TGF-β1) and epithelial growth factor (EGF) on the invasiveness of human laryngeal carcinoma cell line Hep-2 induced by epithelial-mesenchymal transition. Methods The morphological changes of human laryngeal carcinoma cell line Hep-2 were observed 24 h and 48 h after TGF-β1 stimulation and TGF-β1 combined with EGF. Transwell assay was used to detect cell invasion ability and the expression of E-cadherin and vimentin were detected by RT-PCR and Western blot. Results ①TGF-β1 treated 48 h group and TGF-β 1 combined with EGF for 24 and 48 h could both stimulate the cell morphology of Hep-2 cells. ② The invasive ability of Hep-2 cells in both TGF-β1 and EGF treated groups was significantly stronger than that of TGF-β1 treated group (P <0.05) at 24 hours and 48 hours respectively. TGF-β1 alone treated 48 hours Invasion ability was significantly stronger than the control group (P <0.05). ③ The expression of E-cadherin in the group of TGF-β1 combined with EGF decreased but the expression of Vimentin increased obviously. Conclusion EGF can induce epithelial-to-mesenchymal transition in combination with TGF-β1, making laryngeal carcinoma more invasive.