Effect of phosphatase and tensin homolog on chromosome ten engineered mesenchymal stem cells on DBTRG glioma cells: an in vitro study
10.3760/cma.j.issn.1671-8925.2016.12.001
- VernacularTitle:MSC介导的PTEN基因体外作用于人DBTRG胶质瘤细胞的实验研究
- Author:
Xiangjun TANG
1
;
Zhuoshun YANG
;
Li ZHANG
;
Kuanming HUANG
;
Longjun DAI
;
Hanjun TU
Author Information
1. 442000,十堰市太和医院(湖北医药学院附属医院)神经外科
- Keywords:
Mesenchymal stem cell;
Phosphatase and tensin homolog on chromosome ten;
DBTRG glioma cell
- From:
Chinese Journal of Neuromedicine
2016;15(12):1189-1194
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the gene expressions of phosphatase and tensin homolog on chromosome ten (PTEN) engineered mesenchymal stem cells (MSCs) and observe their effects on DBTRG glioma cells.Methods Primary culture of human umbilical cord MSCs was performed;48 h after PTEN transfection into MSCs by liposomes,transfection results were detected under microscope.(1) Transfected MSCsPTEN were used as experimental group and MSCs as control group.PTEN protein and gene expressions of the cells from the two groups were detected by Western blotting and real time-PCR;soluble PTEN protein in the culture supematants was measured using ELISA.(2) In vitro cultured DBTRG cells were divided into four groups,and MSCs supernatant and 25％,50％ and 100％ MSCsPTEN supernatants were added into the four groups,respectively;24 h after culture,calcein AM/EthD-1 staining was used to detect the activity of DBTRG cells which co-cultured with different percentages of MSCsPTEN medium.(3) In vitro cultured DBTRG cells were divided into three groups,and MSCs supernatant and 25％ and 100％ MSCsPTEN supernatants were added into the three groups;RTCA was used to observe the growth curve of DBTRG cells.Results (1) A large number of red fluorescence masses were noted in the MSCsPTEN cells by microscope;real time-PCR and Western blotting indicated that the gene and protein expressions of PTEN in the experimental group were significantly higher than those in the control group (P＜0.05).PTEN content in the supernatants of the experimental group was significantly higher than that in the control group (P＜0.05).(2) CalceinAM/EthD-1 staining indicated that 25％,50％ and 100％ MSCsPTEN supernatant groups had significantly larger number of death DBTRG cells than MSCs supernatant group (P＜0.05);and with the increase of MSCsPTEN supeenatant percentages,the number of death DBTRG cells became larger (P＜0.05).(3) RTCA indicated that as compared with the MSCs supernatant group,25％ and 100％ MSCsPTEN supernatant groups had decreased DBTRG cell index.Conclusion After MSCsPTEN transfection,MSCsPTEN cells can stablely express targeted gene,and their supernatant can obviously promote the death of DBTRG cells and inhibit the growth of glioma cells;PTEN engineered MSCs may be an effective gene therapy for gliomas.
