Phosphatase and tensin homology deleted on chromosome ten inhibition in improving growth of neurons cultured in inhibitive microenvironment
10.3760/cma.j.issn.1671-8925.2014.10.005
- VernacularTitle:抑制PTEN蛋白对抑制性微环境中神经元生长的影响
- Author:
Jianmin FAN
1
;
Jianhong SHEN
;
Fu YAN
;
Fei ZHOU
;
Qi YAO
;
Zhikai GU
Author Information
1. 南通大学附属妇幼保健院神经内科
- Keywords:
Phosphatase and tensin homology deleted on chromosome ten;
Tumor suppressor;
Nerve regeneration;
Primary cell culture
- From:
Chinese Journal of Neuromedicine
2014;13(10):994-998
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of inhibiting tumor suppressor gene phosphatase and tensin homology deleted on chromosome ten (PTEN) on growth of neurons cultured in inhibitive microenvironment.Methods Primary cortical neurons were separated from fetal rats and cultured with adult rat brain-derived myelin membrane proteins as inhibitory substrate.Cells were divided into 3 groups:normal control group,inhibitory control group (myelin membrane proteins as inhibitory substrate) and bpv treatment group (200 nmol/L bpv+myelin membrane proteins).Real-time quantitative-PCR was performed to detect the expressions of PTEN and mammalian target ofrapamycin (mTOR) genes,and immunofluorescence staining was used to observe the growth ofneurites 24,48 and 72 h,and 5 d after inoculation.Results Immunofluorescence staining revealed that significant differences of the growth of neurites at the four observation time points existed among the 3 groups (P＜0.05); during the early days of culture (24,48 and 72 h),cells in normal control group had the longest neurite (43.5±12.2,94.9±23.6 and 154.3±35.4 μm),those in the bpv treatment group enjoyed the second prize (28.8±10.2,75.4±22.5 and 136.8±40.8 μm) and those in the inhibitory control group was obviously inhibited (17.4±5.8,46.3± 13.7 and 78.4 ±29.8 μm).But the difference on the 5th d of cultivation changed into bpv treatment group＞normal control group＞inhibitory control group with the neurite length of (225.9±50.6),(218.4±58.1) and (173.6±48.7) μm,respectively (P＜0.05).PCR revealed no significant difference in expressions of PTEN and mTOR genes of the three groups between each two time points (P＞0.05); while at the same time point,significantly increased expressions ofmTOR and PTEN in the bpv treatment group were noted as compared with those in the normal control group and inhibitory control group (P＜0.05).Conclusions Growth of neurons in inhibitive microenvironment is improved on condition that the tumor suppressor gene PTEN is inhibited,which is probably related to the activation of PI3K/AKT/MTOR pathway.
