| ObjectivesThis study used qrt-PCR and Western blot to detect the expression of EGFR and EZH2 in breast cancer tissues and benign breast tumors of different pathological types,and to explain the expression and correlation of these markers in breast cancer.Clinical and pathological indicators to explore the significance of EGFR and EZH2 in the prognosis evaluation of adenocarcinoma;through bioinformatics software,to find common pathways in which EGFR and EZH2 may play a role,and to provide new possible evaluation basis and therapeutic target for accurate treatment of breast cancer point.MethodsWe collected 120 specimens of breast cancer tissue diagnosed before surgery from September 2017 to September 2018(including Luminal A type,Luminal B Her2 positive type,Her2 overexpression type,TNBC type,30 cases each)and benign breast tumors.qRT-PCR and western blot were used to detect the expression of EGFR and EZH2 mRNA and protein in breast cancer tissues and benign breast tumor tissues.The traditional clinicopathological parameters such as tumor size,lymph node metastasis,clinical stage and histological grade of breast cancer patients were collected,as well as prognostic indicators such as Ki67 and P53 in breast cancer immunohistochemistry.The results were analyzedby statistical software SPSS22.0.The measurement data were expressed as mean±standard deviation.The data between two groups or groups were compared by t test or analysis of variance.The comparison between count data was analyzed by chi-square test or rank sum.Pearson correlation analysis between EGFR and EZH2;P=0.05 was used as the significance test level.The bioinformatics software DAVID 6.7 and KEGG were used to find the common pathways in which EGFR and EZH2 may act,and the results of qRT-PCR and western blot were combined.Results1?The average age of patients with benign breast tumors was 41.41±11.31 years old,and the average age of breast cancer patients was 50.26±10.63.There was a statistically significant difference in age distribution between the two(P<0.05),but there was no significant difference in height and weight(P<0.05).>0.05);2?The mean ages(years)of Luminal type A,Luminal B Her2 positive type,Her2 overexpression type,and TNBC type were 48.10±10.57,51.31±13.03,52.96±8.96,49.03±8.71,and the average body weight(Kg)were 56.74±8.27,respectively.55.77±7.31,56.12±8.81,56.83±9.58,the average height(cm)were156.17±5.39,156.73±7.31,155.38±4.58155.93±6.09,respectively.There was no significant difference in age,height and body weight between different molecular breast cancer patients(P>0.05).The tumor size distribution of breast cancer patients with different molecular classification was compared by chi-square test.Statistical significance(P>0.05);3?The expression rates of EGRF protein in benign breast tumor tissues and breast cancer tissues were 23.3%(7/30)and 45.8%(55/120),respectively.The expression rates of EZH2 protein in benign breast tumor tissues and breast cancertissues were respectively 30%(9/30),65%(78/120).The expression of EGFR and EZH2 protein in breast cancer tissues was significantly higher than that in benign breast tumor tissues,and the difference was statistically significant(P<0.05);The expression levels of EGFR mRNA and EZH2 mRNA in benign breast tumors were 0.471±0.239 and 0.328±0.114,respectively.The expression levels in breast cancer were 0.684±0.317 and 0.493±0.294,respectively.Thus,EZH2 and EGFR in breast cancer tissues were observed.The expression of protein and mRNA was significantly higher than that of benign breast tumor tissues,and the difference was statistically significant(P<0.05).4?In 55 EGFR-positive breast cancer tissues,Luminal type A,Luminal B Her2 positive type,Her2 over-expressed type,and TNBC type accounted for 9cases,11 cases,13 cases,and 22 cases,respectively,and the expression levels were 0.392±0.152 and 0.584±0.217,0.922±0.547,1.474±0.556,respectively.The expression of EZH2 protein in triple-negative breast cancer was significantly higher than that of other types of breast cancer,and the difference was statistically significant(P<0.05).while in non-triple negative breast cancer There was no significant difference in the expression between the any two types(P>0.05).In 78 cases of EZH2 protein-positive breast cancer tissues,Luminal type A,Luminal B Her2 positive type,Her2 over-expressed type and TNBC type accounted for 14 cases,17 cases,19 cases and 28 cases,respectively,and their expression levels were 0.367±0.154,0.628±0.204,0.659±0.227,1.079±0.330 respectively.The expression of EZH2 protein in triple-negative breast cancer was significantly higher than that of the other three types of breast cancer,the difference was statistically significant(P<0.05),while it wasnon-three negative.There was no significant difference in the expression between the two types of breast cancer(P>0.05).5?Luminal A,LuminalB Her2 positive,Her2 overexpression,TNBC breast cancer,EGFR mRNA expression was 0.429 ± 0.350,0.602 ± 0.373,0.805 ±0.130,0.902 ± 0.098,EGFR mRNA in triple negative breast cancer The expression level was significantly higher than that of the other three types of breast cancer,and there was a statistical difference between TNBC and LuminalA,and between LuminalA and Her2 overexpression(P<0.05),but there was no difference between the other two types.Statistical significance(P>0.05);EZH2mRNA expression in Luminal type A,LuminalB Her2 positive type,Her2 overexpression type,TNBC type breast cancer were 0.235±0.115,0.375±0.246,0.559±0.251,0.805±0.191,EZH2 mRNA,respectively.The expression level in triple-negative breast cancer was significantly higher than that in the other three types of breast cancer,and there was a statistical difference between TNBC and LuminalA,LuminalB Her2 positive type,and LuminalA and LuminalB Her2positive(P<0.05).There was no statistical difference between the other two types(P>0.05);6?According to Pearson correlation analysis,there was a positive correlation between EGFR protein and EZH2 protein in breast cancer tissues,r=0.47,P=0.007,showing a moderate correlation;EGFR mRNAwas positively correlated with EZH2 mRNA expression level,r=0.388,P =0.013,showing a low correlation.7 ? The expression levels of EGFR and EZH2 protein and mRNA were correlated with lymph node metastasis and clinical stage(P<0.05),but not with age,menopausal status,tumor size,histological grade and ki-67 protein and P53 protein.(P>0.05),the expression of EZH2 and EGFR protein and mRNA in breast cancer tissues with high number of lymph node metastasis and highclinical stage were increased;8?Bioinformatics analysis showed that EGFR and EZH2 are co-enriched in one molecular function,three biological processes and one cancer miRNA pathway;one of the classical pathways activated by EGFR phosphorylation is PI3 K pathway.HER2 overexpression and an important pathway in TNBC breast cancer;the downstream signaling pathway initiated by EGFR is the FOXO pathway,and EZH2 regulates the expression of FOXC1 gene.Conclusions1?The expression of EGFR and EZH2 in human breast cancer tissues was higher than that in benign breast tumor tissues.The high expression of EZH2 and EGFR may participate in the malignant transformation of breast cancer tissues and promote the progression of breast cancer.2?Among different molecular types of breast cancer,EGFR and EZH2 were the highest in triple-negative breast cancer,and poor prognosis of triple-negative breast cancer may be associated with high expression of EGFR and EZH2.3?The higher the clinical stage and the higher the number of lymph node metastasis,the higher the expression of EGFR and EZH2.The combined detection of EGFR and EZH2 in human breast cancer tissue is expected to be effective method for early diagnosis,targeted therapy and prognosis of breast cancer.4?EGFR,EZH2 may act together in the FOXO signaling pathway,thereby promoting the occurrence and progression of breast cancer,and is associated with poor prognosis in breast cancer,especially in triple-negative breast cancer patients. |
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