| Purpose: Glioma is the most common primary malignant tumor of the central nervous system in adults.In recent years,the incidence of primary glioma has increased year by year,and the annual growth rate has reached 1.2%.One of the most important features of glioma is invasive growth.With the continuous development and large-scale application of high-throughput sequencing technology and gene microarray technology in the field of life sciences,and the in-depth research and application of molecular targeted drug therapy in tumor field,more and more evidence indicates that Lnc RNA is abnormal expression in malignant tumors such as glioma,and regulates the development of tumor through its malignant biological behavior.The length of Lnc RNAs is more than 200 nucleotides,no function coding protein,initially considered genome byproduct of transcription.Multiple Lnc RNAs are found to play the role of oncogene(and)or tumor suppressor genes in glioma,and are closely related to clinical manifestations,diagnosis,molecular targeted therapy,patient reclassification and prognosis.At present,the expression level and possible biological function of long non-coding RNA LINC01116 in human brain glioma are rarely reported.Therefore,we used the combined bioinformatics analysis to screen the long non-coding RNA LINC01116 in glioma;To explore the expression level and relevant clinical significance of LINC01116 in glioma clinical samples and glioma cells level;LINC01116 was down-regulated to study the proliferation,apoptosis,invasion and migration ability,cell cycle ability and the effect on the growth of glioma cell lines in U251,Ln229 and U87;To increase the expression of LINC01116 in U251 glioma cell line,the effect of regulating tumor cell proliferation and invasion and migration was studied.Methods:(1)LINC01116 was screened by chip database(GEO)and sequencing database(TCGA)analysis.TCGA clinical data were used to analyze the relationship between the expression level of LINC01116 and the prognosis of patients with glioma.(2)A total of 27 cases of glioma tissue samples and 10 cases of non-tumor brain tissue samples were collected.The expression of LINC01116 in glioma tissues and non-tumor brain tissues was compared by q RT-PCR,and the correlation analysis was conducted with the collected clinicopathological data.(3)The expression of LINC01116 in U251,Ln229,U87 and SVG cells was compared by q RT-PCR.Experiments were carried out to perform a function deletion and a function acquisition experiment.(4)In vitro,the expression level of LINC01116 was down-regulated in U251,Ln229 and U87 cells lines.CCK8,clone formation and EDU were performed to detect the effection of LINC01116 on proliferation of tumor cells;Transwell was performed to detect the effection of LINC01116 on invasion and migration of tumor cells.In vivo,the expression level of LINC01116 in U87 cells was down-regulated to observe the tumor formation of U87 cells in immunodeficient nude mice.(5)The expression level of LINC01116 was up-regulated in U251 cells by overexpression plasmid;CCK8,clone formation and EDU were performed to detect the effection of LINC01116 on proliferation of tumor cells.Transwell was performed to detect the effection of LINC01116 on invasion and migration of tumor cells.Results: The first Part:(1)The results of bioinformatics analysis indicated that LINC01116 was significantly higher in glioma(P<0.01).Survival analysis suggested that the higher the expression of LINC01116,the more unfavorable the survival of patients.(2)The results of q RT-PCR showed that the expression of LINC01116 in human glioma tissues was significantly higher than that of non-tumor brain tissues,and the results were statistically significant(P<0.01).The clinical pathological grade,tissue type and LINC01116 expression of glioma tissues were positively correlated,and the results were statistically significant(P<0.01).The second part:(1)The results of q RT-PCR showed that the expression of LINC01116 in U251,Ln229 and U87 cells was significantly higher than that of SVG cells,and the results were statistically significant(P<0.01).(2)The expression of LINC01116 was down-regulated in U251,Ln229 and U87 cells,the results of CCK8,clone formation and EDU suggested that the expression of low LINC01116 inhibited the proliferation ability of glioma cells,and the results were statistically significant(P<0.05).(3)The results of cell cycle by flow cytometry analysis indicated that the cell cycle progression was significantly blocked after the expression of low LINC01116 expression and the results were statistically significant(P<0.05);The results of apoptosis by flow cytometry analysis indicated that the percentage of apoptosis of the cells was significantly increased after the expression of low LINC01116 expression,the results were statistically significant(P<0.05).(4)The results of invasion and migration through transwell suggested that after the down-regulation of LINC01116 expression,the invasion and migration ability of U251,Ln229 and U87 cells decreased,the results were statistically significant(P<0.05).(5)In vivo,after the expression of LINC01116 was down-regulated,the tumor growth slowed down in the nude mice with immunodeficiency.The results of q RT-PCR indicated that the expression of LINC01116 in the down-regulated group decreased significantly,which was statistically significant compared with the control group(P<0.01).And the third part:(1)The expression of LINC01116 was up-regulated in U251 cells.(2)The results of CCK8,clone formation and EDU suggested that the increase of LINC01116 expression promoted the proliferation ability of glioma cells,and the results were statistically significant(P<0.01).(3)The results of invasion and migration by transwell indicated that after the increase of LINC01116 expression,the invasion and migration ability of U251 cells was enhanced,and the results were statistically significant(P<0.01).Conclusion: The expression of LINC01116 in human glioma samples is significantly higher than that of non-tumor brain tissues,and the clinical pathology level of glioma tissues are positively correlated with the expression of LINC01116.In glioma cells U251,Ln229 and U87,high expression of LINC01116 can promote tumor cells proliferation,invasion and migration,and inhibit tumor cells apoptosis.LINC01116 plays the role of oncogene in glioma. |
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