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目的:探讨Notch信号通路在人妊娠滋养细胞肿瘤发生发展过程中的重要作用,及其与EMT之间的关系。方法:采用γ-分泌酶抑制剂DAPT处理JAR和JEG-3两种人绒毛膜癌细胞株,MTT法检测两种细胞株的增殖情况,q PCR法检测Notch1 mRNA表达。Western blot法检测DAPT阻断Notch信号通路后JAR细胞中EMT相关蛋白E-cad的表达情况。结果:DAPT能抑制JEG-3、JAR细胞生长,10μmol/L和15μmol/L DAPT作用48h开始分别抑制JEG-3和JAR细胞生长(P<0.05),且抑制作用呈时间和浓度依赖关系。JEG-3细胞中,1μmol/L和10μmol/L DAPT作用组中Notch1 mRNA表达下调,两组比较差异无统计学意义(P>0.05);JAR细胞中,10μmol/L和15μmol/L DAPT作用组中Notch1 mRNA表达下调,两组比较差异有统计学意义(P<0.05)。DAPT能上调JAR细胞中E-cad表达。结论:Notch信号通路和EMT与妊娠滋养细胞肿瘤的发生发展有关;γ-分泌酶抑制剂DAPT或可成为妊娠滋养细胞肿瘤治疗的一个新靶点。
Objective: To investigate the important role of Notch signaling pathway in the development of human gestational trophoblastic tumor and its relationship with EMT. METHODS: Two human choriocarcinoma cell lines, JAR and JEG-3, were treated with γ-secretase inhibitor DAPT. The proliferation of the two cell lines was detected by MTT assay. The expression of Notch1 mRNA was detected by q PCR. Western blot was used to detect the expression of EMT-related protein E-cad in JAR cells after DAPT blocked Notch signaling pathway. Results: DAPT could inhibit the growth of JEG-3 and JAR cells, and inhibited the growth of JEG-3 and JAR cells at 10μmol / L and 15μmol / L DAPT for 48h (P <0.05). The inhibitory effect was dose- and time-dependent. In JEG-3 cells, the expression of Notch1 mRNA was down-regulated in the groups of 1μmol / L and 10μmol / L DAPT, but there was no significant difference between the two groups (P> 0.05). In JAR cells, the effect of DAPT at 10μmol / L and 15μmol / Notch1 mRNA was down-regulated in the two groups (P <0.05). DAPT up-regulates E-cad expression in JAR cells. Conclusion: Notch signaling pathway and EMT are associated with the occurrence and development of gestational trophoblastic tumor. DAPT, a γ-secretase inhibitor, may be a new target for the treatment of gestational trophoblastic tumor.