Study On The Expression And Correlation Of ATRX And TERT In Different Grades Of Glioma

Objective:The expressions of ATRX and TERT in gliomas and normal brain tissue were detected by immunohistochemistry(IHC)and real-time fluorescence quantitative PCR(RT-PCR),combined with gene sequencing to analyze their expression and tumor WHO grade,patient age,patient gender The relationship between the ATRX and TERT promoters in the experimental research and clinical application of gliomas.Methods:1.30 cases of WHO ?,?,and ? grade astrocytoma wax specimens were selected,and 3 normal brain tissue specimens from patients with craniocerebral trauma were made into wax specimens,and sliced for HE staining to compare their tumor characteristics.The expressions of ATRX and TERT were examined by IHC method.Sanger sequencing detected the mutation status of ATRX and TERT promoters.2.Twenty-one clinical specimens of WHO ?,?,and ?astrogliomas and 3 normal brain tissue specimens from patients with traumatic brain injury were selected,and the expression of ATRX and TERT promoters at the nucleotide level was determined by RT-PCR.3.Using IHC method and RT-PCR method to detect the results,SPSS 20.0 and GraphPad Prism 5 statistical analysis were used to compare the expression of ATRX and TERT at the nucleotide level and protein level and the gene mutation status with the tumor WHO grade and patient age And the relationship between gender;comparative analysis of the correlation and degree of ATRX and TERT expression in gliomas.Results:1.IHC and sequencing results:(1)HE staining showed that the cancer cell infiltration in tumor tissue specimens was significantly increased compared with normal brain tissue specimens,and there was obvious heteromorphism;(2)the expression of tumor tissue ATRX and TERT at the protein level Significantly higher than normal brain tissue(P<0.05));(3)The tumor tissues were divided into three groups according to WHO grades ?,?,and ?.The results showed that there was a negative correlation between ATRX mutation and WHO classification(P<0.05),There is a positive correlation between mutations in the TERT promoter and WHO classification(P<0.05);(4)The tumor tissues were divided into three groups A,B,and C according to the age of 35 years old and below,35-54 years old,and 55 years old and above.:There is a negative correlation between ATRX mutation and age classification(P<0.05);there is no significant difference in TERT promoter mutation between different age groups(P>0.05);(5)The tumor tissue is divided into male and female groups according to gender,The results show that:ATRX mutation and TERT promoter mutation have no significant gender difference(P>0.05);2.RT-PCR experimental results:(1)The expression of ATRX and TERT at the nucleotide level of tumor tissue is similar to that of normal brain tissue The ratio was significantly increased(P<0.05);(2)The tumor tissues were divided into three groups according to WHO grades ?,?,and ?.The results showed that there was a positive correlation between ATRX expression and WHO classification(P<0.05),and TERT expression and WHO There is a negative correlation between grades(P<0.05);(3)The tumor tissues are divided into three groups A,B,and C according to the age of 35 years old and below,35-54 years old,55 years old and above.The results show that:ATRX expression and age grade There is a positive correlation(P<0.05);there is a negative correlation between TERT expression and age grading(P<0.05);(4)The tumor tissue is divided into male and female groups according to gender,and the results show that neither ATRX nor TERT expression Obvious gender difference(P>0.05);3.Two methods(IHC and RT-PCR method)in the detection of glioma ATRX and TERT expression data consistency is good(P<0.05);4.ATRX and TERT expression Correlation and degree of correlation:(1)IHC experiment:there is a negative correlation between the expressions of ATRX and TERT(P<0.05);(2)RT-PCR experiment:there is a significant negative correlation between the expressions of ATRX and TERT(P<0.001)Conclusions:1.In gliomas,the expression of ATRX and TERT at both the nucleotide level and the protein level are significantly higher than in normal brain tissue.2.The ATRX mutation rate decreases with the rise of glioma grade.The ATRX mutation is higher in the younger age group,but will decrease with age.The ATRX mutation has nothing to do with gender;the TERT promoter mutation rate varies with the glioma grade Ascending and increasing,the TERT promoter mutation is lower in the younger age group,but will increase with age.The TERT promoter mutation has nothing to do with gender.3.The expression of ATRX increases with the rise of glioma grade,and the expression is lower in the younger age group,but will increase with age.The expression of ATRX has nothing to do with gender;the expression of TERT increases with the rise of glioma grade Decreased,while the expression in the younger age group is higher,but will decrease with age,TERT expression has nothing to do with gender.4.Using IHC method and RT-PCR method to detect the expression of ATRX and TERT in glioma,the results are consistent.5.In gliomas,there may be a negative correlation between ATRX expression and TERT expression.ATRX mutations and TERT promoter mutations are likely to be mutually exclusive.