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用不同浓度(1、4、8、16、32μmol/L)的DNA甲基化酶抑制剂5-Aza-CdR处理绒毛膜癌JEG-3细胞,显微镜下观察细胞形态的变化,MTT法检测5-Aza-CdR对JEG-3细胞增殖的抑制作用,流式细胞术检测5-Aza-CdR对JEG-3细胞凋亡的影响,Transwell小室检测JEG-3细胞侵袭能力的改变,进一步应用Real Time PCR及Western印迹检测5-Aza-CdR处理细胞后DNA甲基化转移酶3a(DNA methyltransferase 3a,Dnmt3a)基因及其蛋白的表达变化。研究结果显示,5-Aza-CdR能致细胞皱缩、细胞核聚集成块,可明显抑制细胞增殖,促进其凋亡,降低细胞侵袭力。5-Aza-CdR同时还可抑制Dnmt3a mRNA和其蛋白的表达。
Choriocarcinoma JEG-3 cells were treated with 5-Aza-CdR, a DNA Methylase inhibitor of different concentrations (1, 4, 8, 16, 32μmol / L). Cell morphology was observed under a microscope. Aza-CdR on the proliferation of JEG-3 cells. Flow cytometry was used to detect the effect of 5-Aza-CdR on the apoptosis of JEG-3 cells. Transwell chamber was used to detect the invasion ability of JEG-3 cells. Real Time The changes of DNA methyltransferase 3a (Dnmt3a) gene and its protein expression after 5-Aza-CdR treatment were detected by PCR and Western blot. The results show that 5-Aza-CdR can cause cell shrinkage, nuclear aggregates into blocks, can significantly inhibit cell proliferation, promote apoptosis, reduce cell invasiveness. 5-Aza-CdR also inhibits Dnmt3a mRNA and its protein expression.