17-烯丙氨基格尔德霉素对人绒癌JAR细胞迁移和黏附的影响

Effects of 17-AAG on Migration and Adhesion of Human Choriocarcinoma JAR Cells

目的探讨17-烯丙胺-17-脱甲氧格尔得霉素(17-AAG)对人绒癌JAR细胞迁移和黏附的影响。方法 MTT实验:以5、10、20、40μg/m L浓度的17-AAG处理绒癌JAR细胞(实验组),设立空白对照组,空白对照组为与实验组相同体积的培养基,培养24 h后观察细胞形态,MTT法检测细胞增殖抑制率,并计算IC50。划痕治愈实验:以5、10μg/m L浓度的17-AAG及空白对照组处理绒癌JAR细胞(实验组),进行细胞划痕治愈实验,划痕后即刻及48 h后,拍照记录细胞划痕治愈情况,并计算划痕治愈率。Transwell实验:以5、10、20、40μg/m L浓度的17-AAG处理绒癌JAR细胞(实验组),设立空白对照组,培养12 h后,Transwell小室迁移实验检测细胞迁移能力。Western blot实验:以5、10、20、40μg/m L浓度17-AAG处理绒癌JAR细胞(实验组),设立空白对照组,培养24 h后,蛋白免疫印迹(Western blot)法检测细胞迁移黏附相关蛋白表达水平。结果 MTT实验结果显示,与空白对照组比较,实验组细胞增殖抑制率明显增加,呈浓度依赖性(P<0.05),24 h后IC50均值为11.951;划痕治愈实验结果显示,与对照组比较,实验组划痕覆盖面积明显降低(P<0.05);Transwell实验结果显示,与空白对照组比较,实验组细胞穿膜数目逐渐降低,呈浓度依赖性(P<0.05);Western blot实验结果显示,与空白对照组比较,实验组E-cadherin蛋白表达水平显著增加(P<0.05),同时N-cadherin、β-catenin蛋白表达水平降低(P<0.05)。结论 17-AAG能够抑制人绒癌JAR细胞增殖,并通过EMT信号通路上调E-cadherin蛋白,下调N-cadherin、β-catenin蛋白的表达抑制人绒癌JAR细胞的迁移和黏附。

Objective To investigate the effect of 17 - AAG on migration and adhesion of JAR cells. Methods JAR cells were cultured in vitro with 17 - AAG at different doses. The inhibitory effects of 17 - AAG was measured by MTr assay for calculation of the IC50. Cells migration and adhesion scratches abilities were assessed by cell scratches method and Transwell chamber migration test. The expression of E - cadherin, N - eadherin and 13 - eatenin was assessed by western blotting analysis. Results The results of MTT assay showed that 17 - AAG significantly inhibited the prolifer- ation of JAR cells in dose - dependent manner （ P 〈 0. 05 ）. The results of scratches method showed that the coverage rate of the experimental group was significantly lower than that of control group （ P 〈 0. 05 ）. The results of Transwell method showed that the number of cell penetrating cells was decreased in dose - dependent manner compared with control group （ P 〈 0. 05 ）. E - cadherin protein expression was significantly increased in experimental group than control group, while N - eadherin and th theon was signifieanreduced （ P 〈 0. 05 ）. Conclusion Our results suggest that 17 - AAG inhibits JAR cells proliferation. It can inhibit JAR migration and adhesion through mediating EMT signaling pathway by decreasingthe expression of N - cadherin and 13 - catenin protein and increasing the expression of E - cadherin. These results indi- cate that 17 - AAG could act as an molecular target and may serve as a promising therapeutic strategy for JAR.