Effect of different concentrations of Netrin-1 on proliferation and apoptosis of TEV-1 cells
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摘要: 目的 探讨不同浓度Netrin-1对滋养细胞株TEV-1增殖凋亡的影响。 方法 行经0、10、100及1 000 ng/ml Netrin-1干预TEV-1 24 h后,采用噻唑兰MTT比色法检测细胞增殖指数、流式细胞法检测细胞凋亡率,采用免疫荧光组织化学方法鉴定TEV-1表达Netrin-1情况,采用实时定量PCR及western印迹分别检测各组细胞Netrin-1mRNA及蛋白变化情况。 结果 以递增浓度的Netrin-1预处理TEV-1细胞24 h后,TEV-1细胞增殖行为无影响,但TEV-1细胞凋亡率随Netrin-1浓度增加而降低,当Netrin-1刺激浓度达1 000 ng/ml时,TEV-1细胞总凋亡率下降至(6.63±0.07)%,较空白组(0 ng/ml)凋亡率(14.44±0.85)%下降明显(P<0.01)。Netrin-1表达于滋养细胞细胞质。在培养的滋养细胞株TEV-1中加入不同剂量的Netrin-1干预24 h后,TEV-1 Netrin-1蛋白表达水平无明显变化。 结论 Netrin-1能明显降低滋养细胞的凋亡程度,期间涉及机制与滋养细胞Netrin-1表达水平无关。Abstract: Objective To explore the role of different dose of Netrin-1 on regulating the proliferation and apoptosis of human trophoblast TEV-1 cells. Methods TEV-1 cells were treated with 0,10,100 and 1 000 ng/ml Netrin-1 for 24 h.Cell viability was measured by MTT assay;cell apoptosis were assessed by flow cytometry;the expression of Netrin-1 in TEV-1cells was observed by indirect cellular immunofluorescence;the mRNA and protein of Netrin-1 expression in TEV-1 cells was performed by real-time PCR and Western blot. Results Immediately after 24 hour of treatment with Netrin-1,the proliferation of TEV-1 cells was independent of Netrin-1,meanwhile,cell apoptosis decreased with the accrescence of concentration.The apoptosis rate significantly decreased to (6.63±0.07)% at 1 000 ng/ml Netrin-1 than (14.44±0.85)% without Netrin-1(P<0.05).Immunoreactivity of Netrin-1 was observed in cytoplasm of trophoblast.The expression amount of Netrin-1 protein in TEV-1 cells was independent of the accrescence of concentration by using Western blot. Conclusion Netrin-1 can reduce cell apoptosis in TEV-1 cells,but the expression level of Netrin-1 may not involve in it.
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Key words:
- Netrin-1 /
- Trophoblast /
- Cell apoptosis /
- Proliferation
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