食管癌相关基因4在体外纯化培养大鼠脉络丛上皮细胞中的表达研究

Expression of esopageal cancer related gene 4 in choroid plexus epithelial cells of purified rat in vitro

摘要:

目的：研究食管癌相关基因4（esopageal cancer related gene 4，ECRG4）在体外培养条件下大鼠脉络丛上皮细胞（choroid plexus epithelial cells，CPECs）中表达的情况。方法：分离纯化培养新生1 d斯泼累格·多雷大鼠（Sprague-Dawley rats，SD Rats）CPECs，并进行传代培养，分为原代培养组（A组）、传1代培养组（B组）和传2代培养组（C组）、原代培养干预组（D组，使用氯化锰给予毒化），并应用qRT-PCR法、Western blot法及ELISA法分别检测ECRG4在各组CPECs中的表达情况。结果：①qRT-PCR法检测ECRG4 mRNA在原代培养和传代培养的CPECs中的表达：B组ECRG4 mRNA表达高于A、C 2组，差异具有统计学意义（P=0.000），A、C 2组差异无统计学意义（P=0.127）。D组与A组比较表达明显降低，差异有统计学意义（P=0.001）。②Western blot法ECRG4蛋白在原代培养和传代培养的CPECs中的表达：B组ECRG4蛋白表达水平明显高于A组及C组，C组ECRG4蛋白表达水平明显高于A组，差异具有统计学意义（P=0.000）。A组ECRG4蛋白表达水平明显高于D组，A、D 2组ECRG4蛋白表达水平的差异具有统计学意义（P=0.000）。③ELISA方法检测ECRG4在原代培养和传代培养的CPECs中的分泌情况：B组高于A、C 2组，A组高于C组，差异具有统计学意义（P=0.000）。而A、B 2组差异无统计学意义（P=0.131）。结论：通过对ECRG4在CPECs中的表达研究，提示ECRG4可能参与CPECs对损伤的反应，并影响了CPECs的生物学特性，为ECRG4在神经再生中的作用研究奠定理论和实验基础。

Abstract:

Objective：To detect expression of esopageal cancer related gene 4（ECRG4） in choroid plexus epithelial cells of purified rat in vitro. Methods：Primary and passage choroid plexus epithelial cells were obtained from 1 d newborn Sprague-Dawley rats. Expres－sion of ECRG4 was measured by qRT-PCR and Western blot. Secretion of ECRG4 was detected by ELISA. Cells were divided into primary cells（A），passage 1 cells（B） and passage 2 cells（C） groups. Cells which using MnCl2 were divided into intervening primary cells（D） groups. Results：①Expression of ECRG4 mRNA was higher in group B than in groups A and C with statistical differences（P=0.000）. There was no statistical difference in expression of ECRG4 mRNA between group A and group C（P=0.127）. Expression of ECRG4 mRNA was significantly reduced in group D than in group A（P=0.001）. ②ECRG4 protein expression was significantly higher in group B than in group A and group C. ECRG4 protein expression was significantly higher in group C than in group A（P=0.000）. ECRG4 protein expression was significantly higher in group A than in group D（P=0.000）. ③Secretion of ECRG4 detected by ELISA was significantly higher in group B than in group A and group C. Secretion of ECRG4 was significantly higher in group A than in group C（P=0.000）. There was no statistically significant difference between group A and group B（P=0.131）. Conclusions：ECRG4 might participate in the responding of choroid plexus epithelial cells to injury and influence bionomics of choroid plexus epithelial cells. The work on ECRG4 expression in choroid plexus epithelial cells can establish theoretical and experimental basement for clarifying the function of choroid plexus epithelial cell in nerve re－generation promoting.