轴突导向因子netrin-1在滋养层细胞侵袭中的作用  

Correlation of netrin-1 expression with invasion of extra villous trophoblasts

作　　者：杨昀[1] 王勇[1] 杜燕[1] 朱剑文[1] 高慧[1] 邹丽[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院妇产科,武汉430022

出　　处：《中华妇产科杂志》2009年第2期131-134,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的探讨轴突导向因子netrin—1在滋养层细胞侵袭中的作用及其机制。方法采用RT—PCR技术检测绒毛膜外滋养细胞株TEV-1中netrin-1的6种受体（UNC5A、UNC5B、UNC5C、UNC5D、DCC和neogenin）mRNA的表达。将培养的细胞按照加入netrin-1蛋白浓度的不同分为10ug/L组、50ug/L组、100ug/L组、500ug/L组、1000ug/L组、5000ug/L组和阴性对照组（netrin-1的浓度为0ug/L）共7组，分别用细胞计数试剂盒8检测各组细胞的增殖能力[以吸光度（A）值表示]，体外侵袭实验检测各组细胞的侵袭能力。结果（1）RT-PCR技术检测结果显示，netrin-1的6种受体中，仅检测到neogenin和UNC5B mRNA的表达。（2）培养72h后，10ug/L组、50ug/L组、100ug/L组、500ug/L组、1000ug/L组和5000ug/L组细胞的A值分别为1．55±0.29、1．72±0．31、2.15±0．35、1．42±0．25、1．50±0．27和1．38±0．23，分别与阴性对照组（1．00±0．16）比较，差异均有统计学意义（P〈0．05）。（3）培养6h后，10ug/L组、50ug/L组、100ug/L组、500ug/L组、1000ug/L组和5000ug/L组细胞的穿膜细胞数分别为（41±4）、（47±5）、（55±6）、（44±5）、（43±5）和（42±5）个，分别与阴性对照组[（30±4）个]比较，差异均有统计学意义（P〈0．05）。（4）neogenin mRNA的表达水平，10ug/L组、50ug/L组、100ug/L组、500ug／L组、1000ug／L组和5000ug/L组分别为1．50±0．16、1．83±0．19、2．24±0．25、2．12±0．24、2．12±0．23和2．13±0．23，分别与阴性对照组（1．00±0．11）比较，差异均有统计学意义（P〈0．05）；10ug/L组与50ug/L组比较，50ug/L组与100ug/L组比较，差异也均有统计学意义（P〈0．05）；100ug/L组与500ug/L组、1000ug/L组和5000ug/L组之间两两比较，差异均无统计学意义（P〉0．05）。（5）UNC5B mRNA的表达水平，10ug/L组、50ug/L组、100ug/L组、500ug/L组、1000ug/L组和5000ug/L组分别为1．09±0．11、1．47±0．14�Objective To investigate mechanism of netrin-1 regulating invasion of extra villous trophoblasts. Methods RT-PCR was used to detect six receptors expression including UNC5A, UNC5B, UNCSC, UNC5D, DCC and ueogenin in extra villous trophoblast cell line TEV-1. The TEV-1 cells were cultured and devided into seven groups according to the concentration of netrin-1 adding into the medium, which include 10 ug/L, 50 ug/L, 100 ug/L, 500 tug, 1000 tug, 5000 ug/L and the control（the concentration of netrin-1 was 0ug/L） groups. The proliferation and invasion of TEV-1 induced by netrin-1 were determined by CCK-8 assay and transwell invasion assay respectively. Results （ 1 ） Only neogenin and UNC5B were found to be expressed on TEV-1 by RT-PCR method. （2） In CCK-8 proliferation assay, after 72 hours culture, the proliferation of TEV-1 were 1.55 ±0. 29 in 10 ug/L, 1.72 ±0. 31 in 50 ug/L, 2. 15 ±0. 35 in 100 ug/L, 1.42 ±0. 25 in 500 ug/L, l. 50 ±0. 27 in 1000 ug/L, and 1.38 ±0. 23 in 5000ug/L group, which were all higher than 1.00 ± 0. 16 in control group significantly （ P 〈 0. 05 ）. （ 3 ） In matrigel invasion assay, after 6 hours culture, the number of the trans-membrane cells in various netrin-1 group, including 41 ±4 in 10 ug/L, 47 ±5 in 50 ug/L, 55 ±6 in 100 ug/L, 44 ±5 in 500 ug/L, 43 ±5 in 1000 ug/L and 42 ± 5 in 5000 ug/L group, were all higher than 30 ± 4 in control group with statistical significance（ P 〈 0. 05 ）. （4） The fold changes of neogenin were 1.50 ± 0. 16 in 10 ug/L, 1.83 ± 0. 19 in 50 ug/L, 2. 24 ±0.25 in 100 ug/L, 2. 12 ± 0. 24 in 500 ug/L, 2. 12 ± 0. 23 in 1000 Iug and 2. 13 ± 0. 23 in 5000 ug/L group, which were all higher than 1.00 ±0. 11 in control group significantly（P 〈0. 05）. There were significant difference between group 10 tug and 50 ug/L, group 50 iug and 100 ug/L （ P 〈 0. 05 ）. There were no significant difference between group 100 Iug/L and 500 tug, group 1000 tug and 5000 tug （P 〉0. 05）. （5）The fold changes of UNC5B 1.09 �

关 键 词：滋养层 神经生长因子类 肿瘤抑制蛋白质类 受体 细胞表面 

分 类 号：R686[医药卫生—骨科学]