人舌鳞状细胞癌顺铂耐药细胞发生上皮-间质转化的研究

Epithelial-mesenchymal transition is associated with chemoresistance in human tongue squamous cell carcinoma cells

目的初步探讨人舌鳞状细胞癌(TSCC)化疗耐药与上皮-间质转化(EMT)的关系。方法以人舌鳞状细胞癌细胞、SCC15及其顺铂耐药细胞株CAL27/CDDP、SCC15/CDDP为研究对象。MTT检测两组亲本细胞及耐药细胞的半抑制率(IC50),显微镜下观察两组亲本细胞及耐药细胞的形态,免疫荧光和Weston blot检测EMT相关分子标记蛋白E-cadherin和Vimentin的表达,Weston blot检测EMT转录因子Twist1和Slug的表达,Transwell及划痕实验检测细胞的侵袭迁移能力,结果采用单因素方差分析。结果 CAL27/CDDP较CAL27 IC50提高7.5倍(χ2=13.8043,P<0.01),SCC15/CDDP较SCC15 IC50提高8.3倍(χ2=10.464,P<0.01)。CAL27和SCC15均为上皮细胞形态,CAL27/CDDP和SCC15/CDDP呈间质细胞形态,且两组耐药细胞上皮标记蛋白E-cadherin表达下调,间质标记蛋白Vimentin表达上调,EMT转录因子Twist1、Slug表达上调,细胞的侵袭迁移能力明显增强。结论顺铂能成功诱导人舌鳞癌细胞发生EMT。

Objective The aim of this study was to make a preliminary exploration on the relationship between chemoresistance of human tongue squamous cell carcinoma （TSCC） and Epithelial- mesenchymal transition. Methods A-MTr-based method was used to analyze the half maximal inhibitory concentration （IC50） values of TSCC cell lines CAL27, SCC15 and their eisplatin-resistant cell lines CAL27/ CDDP, SCC15/CDDP. The morphology change was detected under microscope. Immunofluorescence staining and western blot were used to detect the expression of EMT molecular markers E-cadherin and Vimentin. Western blot was used to detect the expression of EMT transcription factors Twistl and Slug. Transwell assay and scratch test were used to detect the invasion and migration capabilities of cisplatin- resistant cell lines and their parent cell lines, one-way ANOVA was used to analyzed the result. Results The IC50 of CAL27/CDDP increased by 7.5 fold （X^2=13.8043,P 〈 0.01） and that of SCC15/CDDP increased by 8.3 fold（X^2=10.464,P〈 0.01） compared to their parent cell lines. Both CAL27 and SCC15 showed typical epithelial morphology, while CAL27/CDDP and SCC15/CDDP showed mesenchymal morphology. The expression of E-eadherin was down-regulated and that of Vimentin, Twistl and Slug was up-regulated in CAL27/CDDP and SCC15/CDDP ceils compared with their parent cell lines. CAL27/CDDP and SCC15/CDDP ceils showed higher invasion and migration capabilities compared with their parent cell lines. Conclusion Cisplatin induce EMT in human TSCC cells.