Objective To study whether transient cisplatin treatment induces epithelial- mesenchymal transition (EM T) in cervical cancer cells ,and to elucidate the role of Twist 1 ,the master regulator of EMT ,in this process .Methods In HCE1 cells ,EMT after cisplatin treatment was examined in vitro by inverted phase contrast microscope ,MTT assay ,immunohistochemical method and transwell chamber migration assay at 24 ,48 ,72 ,96h ,to examine morphologic changes ,growth trends ,expression of protein and malignant phenotype respectively .The cisplatin-induced Twistl expression was examined by semi-quantitative RT-PCR .Results Low dose cisplatin induced EMT in morphology of HCE 1 cells after 72 h .The expression of E-cadherin was decreased ,the expression of Vimentin and P-gp were all enhanced .The average number of penetrating cells in the control group and EMT group were (39 .000 0±2 .828 4) and (100 .166 7±3 .868 7) respectively .The OD ratio of Twistl / internal reference measured by RT-PCR method was 0 .200 2±0 .000 9 in the control group and 0 .615 6±0 .005 1 in the EMT group ,the difference was statistically signifi-cant(P<0 .05) .Conclusion Low-dose cisplatin could induced EMT in early stage .The invasive capacity in vitro and drug resistance of HCE1 cells with EMT occurrence are all increased .Twist 1 is the key regulator of cisplatin-induced EMT of cervical cancer cells .%目的 研究短时间应用顺铂是否会引起宫颈癌细胞株HCE1发生上皮间质转化(EMT),明确Twist1是否为调控顺铂诱导宫颈癌细胞EMT的关键转录因子.方法 采用倒置相差显微镜、四甲基偶氮唑盐(MTT)法和免疫组织化学法分别从细胞形态、生长曲线和标志性蛋白E-cadherin、vimentin的表达情况.以HCE1细胞株为对照组,加顺铂组(终浓度0.1 mg/L)为实验组,观察加入顺铂24、48、72、96 h后,宫颈癌细胞株HCE1是否发生EMT及其时间点.采用Transwell小室实验检测肿瘤细胞体外侵袭能力,免疫组织化学方法检测P糖蛋白(P-gp)蛋白表达证实EMT发生.半定量RT-PCR方法比较EMT前、后Twist1表达.结果 低剂量顺铂处理HCE1 72 h后,HCE1细胞形态发生EMT的改变;上皮细胞标志性蛋白细胞黏附分子(E-cadherin)表达减少、间质细胞标志性蛋白Vimentin表达增强,P-gp蛋白表达增强;对照组HCE1平均穿膜细胞数为(39.000 0±2.828 4)个,EMT组HCE1平均穿膜细胞数为(100.166 7±3.868 7)个;RT-PCR法测得Twist1/内参OD比值在对照组0.200 2±0.000 9,EMT组0.615 6±0.005 1,组间比较差异有统计学意义(P＜0.05).结论 低剂量顺铂早期即可诱导HCE1发生EMT,发生EMT的HCE1细胞体外侵袭能力和耐药性增强,Twist1是调控顺铂诱导宫颈癌细胞EMT的关键转录因子.
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