Expression Of GAS41in Glioma And Build GAD41RNA Interference Lentiviral Vector

Objective:To detect the expression of the glioma amplified sequence41(GAS41) gene in different pathological grades of glioma specimens and glioma cell lines, to explore the role of GAS41gene in glioma development.Methods:(1) Immunohistochemical staining and Western blot were used to detect the expression of of GAS41protein in the30cases of gliomas specimens of different pathological grades.(2) Western blot was used to detect the expression of of GAS41protein in the glioma cell lines, and molecular cloning technique were used to build the GAS41RNA interference lentiviral GV118-GAS41-RNAi, the GV118-GAS41-RNAi、pHelper1.0and pHelper2.0vectors were cotransfected into293T incasing cells. Harvesting recombinant lentivirus and detecting viral titer, and the recombinant lentivirus was able to express lentiviral vector, which targeting interference GAS41gene.Results:(1) The expression of GAS41protein in the high-grade glioma was stronger than in the low-grade glioma, and the difference was statistically significant (P=0.011<0.05).(2) The expression of GAS41protein in the A172cell is higher, and the recombinant lentiviral vector GV118-GAS41-RNAi targeting interference GAS41gene was successfully constructed and identified by PCR and DNA sequencing. GV118-GAS41-RNAi、pHelper1.0and pHelper2.0vectors were cotransfected into293T cell lines, the recombinant lentivirus was harvested from supernatant of293T cells with a viral titer of5×108TU/ml.Conclusion:(1) The expression of GAS41protein in gliomas is enhanced with the increasing degree of malignancy, which suggests that the expression of GAS41may be used as a criterion of the high level of glioma.(2) The lentivirus interference vector of GAS41was constructed successfully, the A172cell can be used for futher study of interference GAS41gene expression on the biological activity of glioma cells, which lay the foundation for the follow study.