Objective To investigate the effect of soluble Netrin-1 receptor on proliferation and apoptosis of human tro-phoblast TEV-1 cells. Methods Cultured TEV-1 cel s were treated with different concentrations of Neogenin (0, 1, 5, 10 and 50ng/ml). After 24h of treatment cel viability was measured by MTT assay;cel apoptosis were assessed by flow cytometry (FCM) assay;the expression of Netrin-1 in TEV-1 cel s was observed by indirect cel ular immunofluorescence;the mRNA and protein of Netrin-1 were detected by real-time PCR and Western blot, respectively. Results The results of MTT and FCM showed that proliferation of TEV-1 cel s was independent of Neogenin. Meanwhile, cel apoptosis increased with the increasing concentrations of Neogenin;the apoptosis rate was 18.70%at 0 ng/ml Neogenin and significantly increased to 56.65%at 50ng/ml Neogenin (P<0.01). Immunoreactivity of Netrin-1 in cytoplasm of TEV-1 cel s was increased along with the increasing concentration of Neogenin. Real-time PCR showed the level of Netrin-1 mRNA, increased by 40.38 times compared to control group when TEV-1 cel was exposed to 50ng/ml Neogenin(P<0.05);and the same tendency was not seen by using Western blot(P>0.05). Con-clusion The soluble Netrin-1 receptor Neogenin enhances Netrin-1 mRNA expression and promotes cel apoptosis in TEV-1 cel s, which indicates that Netrin-1/Neogenin pathway may play a role during regulating cell apoptosis of trophoblasts.% 目的观察可溶性Netrin-1受体对人滋养细胞增殖凋亡的影响,探究Netrin-1/Neogenin通路在滋养细胞凋亡过程中的机制.方法行早孕期滋养细胞株TEV-1培养,细胞预处理24h后,改用含不同浓度Neogenin的培养液,以浓度为0ng/ml的培养液为对照组,其余不同浓度Neogenin(1、5、10、50ng/ml)干预的培养液均为实验组.应用噻唑蓝(MTT)比色法、流式细胞术(FCM)检测细胞增殖指数、凋亡率；采用细胞免疫荧光法检测Netrin-1蛋白在TEV-1细胞内的表达,real-time PCR及Western印迹检测Netrin-1 mRNA及Netrin-1蛋白表达情况.结果不同浓度的Neogenin对TEV-1细胞增殖无明显影响,但TEV-1细胞凋亡率随 Neogenin浓度的增加而增加.Netrin-1表达于滋养细胞胞质.实验组当 Neogenin浓度达到50ng/ml时,TEV-1细胞表达Netrin-1 mRNA水平较对照组增加约40.38倍,两组间差异有统计学意义(P<0.05).Netrin-1蛋白表达情况受Neogenin浓度影响不明显(P>0.05).结论可溶性Netrin-1受体能促进TEV-1细胞凋亡及Netrin-1 mRNA表达,Netrin-1/Neogenin通路参与了TEV-1细胞凋亡过程.
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