Effects Of FAS-AS1 On Proliferation,migration,and Invasion Of Glioma Cells

Objective : Glioma is a common malignant tumor affecting the central nervous system,accounting for about 40% of intracranial tumors.In 2016,the classification of central nervous system tumors,which integrates histological and genomic phenotypes,was introduced for the first time by the World Health Organization.Gliomas are divided into four grades: grade I is benign glioma,II grade is low grade malignant glioma,and III-IV grade is highly malignant glioma.Glioblastoma(GBM)is the most common and fatal primary brain tumor.Surgical resection plus radiotherapy and chemotherapy are the most common clinical treatments for gliomas.Even under the current ideal conditions,the recurrence rate and mortality rate of glioma patients are still high,the prognosis is generally poor,and the median survival time is less than one year.Therefore,understanding the molecular mechanism of glioma,looking for biomarkers for early diagnosis and exploring effective treatment targets are of great clinical significance to conquer glioma.In recent years,more and more studies have shown that epigenetic regulation,especially long-stranded non-coding RNA,plays an important role in gliomas.Long-strand non-coding RNAFAS-AS1 has been previously reported in breast cancer,non-small cell lung cancer and B lymphoma.However,the study of FAS-AS1 in glioma cells has not been reported.In order to explore the effect of FAS-AS1 on glioma cells,we took clinical glioma tissue specimens and human glioma cell lines U251 and SF126 cells as research objects to explore the effect of FAS-AS1 on the proliferation,migration and invasion of glioma cells.Methods :(1)RNA,extracted from clinical glioma patients and normal brain tissue samples were used to detect the expression of FAS-AS1 in the two groups by real-time quantitative reverse transcription-polymerase chain reaction(q RT-PCR).(2)The expression of FAS-AS1 in different malignant glioma cell lines was detected by q RT-PCR.(3)FAS-AS1 was used to transfect glioma cell lines SF126 and U251,and q RT-PCR was used to verify the transfection efficiency.The cell model of FAS-AS1 overexpression was successfully constructed.(4)The effect of FAS-AS1 on the proliferation of glioma cells was detected by MTT and clone formation assay.(5)The effect of FAS-AS1 on the migration ability of glioma was detected by Transwell experiment.(6)The biofilm constructed by matrix glue combined with Transwell experiment was used to detect the effect of FAS-AS1 on the invasive ability of glioma cells.(7)Western blotting(Weatern Blot)was used to detect the effect of FAS-AS1 on the expression of glioma cell proliferation and invasion related proteins.Result :(1)Compared with normal brain tissues,the expression of long chain noncoding RNA FAS-AS1 in different grades of human gliomas decreased.(2)The expression of long-chain non-coding RNA FAS-AS1 decreased with the increase of glioma grade.(3)The glioma cell model with overexpression of FAS-AS1 was successfully constructed by transfection of FAS-AS1 plasmid.(4)The overexpression of long chain non-coding RNA FAS-AS1 can inhibit the proliferation,migration and invasion of glioma cells.(5)Overexpression of FAS-AS1 could inhibit the expression of proliferation-related protein,proliferating cell nuclear antigen(PCNA)and G1 ram Sspecific cyclin-D1(Cyclin D1),migration and invasion-related protein,matrix metalloproteinase-9(MMP9).Conclusion : All the experimental data show that the expression of FAS-AS1 is low in glioma cells,and the lower the expression is with the increase of glioma grade.Overexpression of FAS-AS1 can inhibit the proliferation,migration,invasion and other cellular biological functions of glioma cells.The expression level of glioma cell proliferation,migration,invasion and other related proteins also decreased with the overexpression of FAS-AS1,which may provide a new direction for targeted therapy of glioma in the future.