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目的:探讨不同浓度Neogenin对滋养细胞侵袭能力改变的影响。方法:行早孕期滋养细胞株TEV-1培养,并随机分为以无血清培养基处理的对照组及含不同浓度Neogenin(1~50 ng/ml)的无血清培养基处理的实验组,培养24 h后利用细胞免疫荧光法行滋养细胞MMP-9定位检测,采用改良Boyden小室法监测不同浓度Neogenin刺激后滋养细胞侵袭能力的变化。利用蛋白免疫印迹技术检测不同浓度Neogenin刺激后滋养细胞MMP9蛋白表达变化情况。结果:MMP-9表达于滋养细胞胞浆。在培养的滋养细胞中加入不同剂量的Neogenin干预滋养细胞24 h后,滋养细胞MMP-9表达量均下降,当Neogenin的干预剂量达到5 ng/ml时,穿膜成功细胞数明显减少(P<0.05)。结论:Neogenin可能通过影响TEV-1细胞MMP-9活性的改变而导致其侵袭能力变化。
Objective: To investigate the effect of different concentrations of Neogenin on the alteration of trophoblast invasion. Methods: The gestational trophoblast cell line TEV-1 was cultured and randomly divided into the control group treated with serum-free medium and the experimental group treated with serum-free medium containing different concentrations of Neogenin (1 ~ 50 ng / ml) Twenty-four hours later, the cells were immunofluorescently assayed for the location of MMP-9 in the trophoblast cells. The modified Boyden chamber method was used to detect the changes of trophoblast invasion after stimulation with different concentrations of Neogenin. Western blotting was used to detect the changes of MMP9 protein expression in trophoblasts after stimulation with different concentrations of Neogenin. Results: MMP-9 was expressed in trophoblastic cytoplasm. Neutrinin intervention of trophoblasts in cultured trophoblasts for 24 h decreased the expression of MMP-9 in trophoblast cells. When Neogenin intervention dose reached 5 ng / ml, the number of successful transplants was significantly decreased (P < 0.05). CONCLUSION: Neogenin may affect the invasiveness of TEV-1 cells by affecting the activity of MMP-9.