| BACKGROUND AND OBJECTIVE: Glioma is the most common intracranial malignant tumor with high mortality and mobility.Long non-coding RNAs(LncRNA)is a kind of non-coding RNA whose length is more than 200 nucleotides,which plays an important role in malignant tumors and may become a key molecular marker of glioma.Searching key LncRNA in glioma is one of the hotspots in basic research of glioma.Current studies have shown that glioma is a group of heterogeneous tumors with various molecular mechanisms.The treatment and prognosis of glioma with different molecular mechanisms are quite different.Therefore,exploring the molecular mechanism of glioma,explaining the genesis and development of glioma at the gene level,and classifying and treating glioma with molecular pathological characteristics are the key directions of clinical research of glioma at present.In this study,the expression profile of LncRNA was constructed by using LncRNA microarray,key LncRNA was screened,and biological function assay were carried out in cell lines and glioma samples.Meanwhile,we analyzed the molecular pathological classification of glioma retrospectively and constructed a new risk stratification of glioma based on molecular pathology to evaluated the prognosis of glioma.Research contents and methods: In the first section,we used 9 glioma specimens and3 normal brain tissues to analyze the expression profile of LncRNA,searching the differentially expressed LncRNA in normal tissues and glioma specimens,and carried out relevant bioinformatics analysis.In the second section,the function and clinical significance of LncRNA MCM3AP-AS1 were studied.First,U251 cells were transfected with MCM3AP-AS1 overexpression vector.After successful transfection,the glioma cell lines expressing MCM3AP-AS1 and the control group transfected with empty plasmid were used to carry out cloning formation test,CCK-8 cell proliferation test,Transwell cell invasion and migration test,and flow cytometry was used to detect the change of cell cycle.MCM3AP-AS1 was detected in 55 glioma specimens.The expression level of MCM3AP-AS1 and its relationship with clinicopathological features and prognosis of glioma were analyzed.The relationship between MCM3AP-AS1 expression and clinical and prognosis of glioma was analyzed by using case and chip data from TCGA and CGGA databases.In the third section the molecular pathology of glioma cases from January 2016 to December2018 were retrospectively analyzed,focusing on the expression of IDH gene status,1p/19 q loss of heterozygosity in diffuse glioma and their clinical significances,and reviewed the special types of glioma,and explored the clinical practice of molecular classification of glioma.In the fourth part,based on the new molecular pathological features,WHO classification,surgical resection and post-operative treatment,the diffuse glioma was re-stratified and divided into low-risk group,medium-risk group,high-risk group and extremely high-risk group.According to the risk stratification criteria,110 cases of diffuse glioma with complete molecular pathological data from2016 to 2018 were retrospectively analyzed.Clinical cases from CGGA database were also used to validate the new risk stratification.RESULTS: There were 185 LncRNAs with significant differences in glioma,90 of which were highly expressed and 95 were low expressed.Bioinformatics analysis showed that these differentially expressed LncRNAs had extensive network connections with micro RNAs and RNA,and participated in signal pathways of cell growth and apoptosis.U251 cells were transfected with MCM3AP-AS1 overexpression vector.Overexpression of MCM3AP-AS1 in U251 cell line suppressed tumor cell proliferation,migration and invasion,and induced apoptosis.The expression of MCM3AP-AS1 was confirmed in 55 glioma specimens.The low expression of MCM3AP-AS1 in glioma tissue specimens was associated with the higher WHO grade and worse prognosis of,while low expression of MCM3AP-AS1 were independent factors of poor prognosis.The analysis of TCGA and CGGA databases confirmed that MCM3AP-AS1 was associated with poor prognosis of glioma.The analysis of CGGA databases further confirmed that the decrease of MCM3AP-AS1 expression was more common in IDH wild-type tumors and related to molecular typing of tumors.This study retrospectively analyzed 188 cases of glioma.Using IDH mutation and 1p/19 q FISH,glioma can be divided into diffuse glioma and other types of glioma.Molecular pathology has important clinical significance in various subtypes of glioma.110 cases of diffuse glioma were divided into four groups by new risk stratification: 24 cases in low-risk group,with a median follow-up period of 17.6 months without death;12 cases in medium-risk group,with an median follow-up period of 18.4 months,only 1 case died;15 cases in high-risk group,with a total survival period of 31.7 months;59 cases in extremely high-risk group,with a median overall survival period of 11.5 months.CGGA database also used risk stratification: 101 cases in low-risk group,median follow-up time was 70.4 months,only 27 cases died,5-year survival rate was 78.38%;152 cases in middle-risk group,median overall survival period was 68 months,5-year survival rate was 53.93%;46cases in high-risk group,median overall survival period was 22.5 months,5-year survival rate was 23.44%;180 cases in extremely high-risk group,median overall survival period was only 12 months,with 5-year survival rate of 11.14%.CONCLUSION: There are a lot of differentially expressed LncRNA in normal tissues and tumor tissues.The construction of differentially expressed LncRNA profiles can provide a basis for searching for key molecular markers.LncRNA MCM3AP-AS1 is a differentially expressed LncRNA,which plays a role in inhibiting tumor proliferation in U251 cell line.Low expression of MCM3AP-AS1 is associated with high grade and poor prognosis of glioma.The molecular classification of glioma can be based on IDH mutation and 1p/19 q LOH.Molecular pathological classification has become an important basis for the classification of glioma.Risk stratification based on molecular pathology and clinical conditions can be used to judge the overall prognosis of diffuse glioma. |
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