Discordance In The ER,PR And Her-2between Primary Tumor And Paired Axillary Lymph Nodes Metastasis Of Triple Negative Breast Cancer

Objective:Breast cancer is one of the most serious cancer problems currently facing women's health.About 100,000 people die of breast cancer each year.Compared with the data of developed countries,although the current incidence rate is still relatively low,the current incidence rate has continued to increase,ranking first in female cancer.Breast cancer,as a common tumor in women,has a high degree of heterogeneity^[1-2].There is a significant difference in terms of molecular,tissue,or response to treatment.Scientific understanding can achieve this difference.Targeted treatment.With the current deeper understanding of molecular typing,triple-negative breast cancer?TNBC?as an independent subtype has gained increasing attention.TNBC is a special type of breast cancer that belongs to both ER and PR receptors and corresponding to the negative category.The proportion in all breast cancers is about 10%-20%.As a group,patients with triple-negative tumors have a relatively poor outcome because it cannot be treated with endocrine therapy or therapies targeted to human epidermal growth factor receptor2.Breast cancer is a kind of higly hetrongenous diseases,not only on the molecular,but also on the biology charaters and the reaction of therapies,even between primary tumor and paired axillary lymph node metastasis.The heterongenous or discordance is also found in TNBC.Can it impact the therapy and prognosis of TNBC?Methods:Seventy cases of breast cancer admitted to the First Affiliated Hospital of China Medical University from 2004 to 2009 were retrospectively collected,all of them were female,and ER and PR were negative with axillary lymph node metastasis confirmed by pathology.The patients did not undergo radiotherapy and chemotherapy before operation.The primary focus and lymph nodes were removed by operation,and subsequent embedding treatment was carried out.Methods:1.HE staining;2.Immunohistochemical SP staining;3.Fluorescence immunoin situ hybridization;4.RT-PCR;5.Western blot;6.Web database.Follow-up:All 70 patients were followed up by telephone.The follow-up deadline was September 2013.The median follow-up lasted80 months.During follow-up,28 patients suffered from recurrence,metastasis and death.Of all the cases,25 died of breast cancer,while 3 died of related cardiovascular problems or traffic accidents.Those who died of other causes during follow-up,lost interviews or survived until the end of the study were counted as censored data.The actual concept of disease-free survival?DFS?is to start at the time after surgery,to follow-up recurrence or metastasis;the specific concept of total survival?OS?belongs to the beginning of the initial diagnosis,to the end of the death time.Statistical processing:SPSS 18.0 and prism7.0 software were used for data processing.The data analysis program includes basic descriptive statistics,chi-square test,t-test and multivariate analysis,which are expressed by the Cox proportional risk regression model.Survival period is calculated by the method followed up,while in the process of inter-group analysis,it is tested by the method.The difference in the generation P<0.05 table has certain statistical value.Result:1.Median follow up was 80 months.Discordance between primary tumors and paired lymph node metastases was 21.4%?15/70?for ER and PR,5.7%?4/70?for.2.Statistical significant of DFS was not found between lymph nodes ER+and ER-groups??2=0.2094,P=0.6472,?2=0.3205,P=0.5713?3.Statistical significance of OS and DFS was not found between lymph nodes PR+and PR-groups??2=0.7218,P=0.3956,?2=0.6314,P=0.4268?4.Statistical significance of OS and DFS was found between lymph nodes+and-groups??2=16.2313,P<0.0001,?2=22.0983,P=<0.0001?5.Cox multivariable analysis revealed metastatic lymph nodes number is independent factor of DFS6.Cox analysis shows the number of transitions and positives as an important independent factor in OS7.The expression of GATA3 in metastatic lymph nodes of 70 patients was detected by immunohistochemistry.Among the 15 patients with ER positive,12 were GATA3positive,the positive expression rate was 80%.Among the patients with ER negative,11were GATA3 positive,the positive rate was 20%.Fisher's exact test showed that there was a significant difference between them?P<0.01?.8.ER expression in ER positive metastatic lymph nodes was significantly higher than that in primary and ER negative metastatic lymph nodes?P=0.0002?.9.The expression level of GATA3 in ER-positive metastatic lymph nodes was significantly higher than that in primary and ER-negative metastatic lymph nodes?P<0.0001?.10.RT-PCR showed that the expression of microRNA-519a in ER-positive metastatic lymph nodes was significantly lower than that in primary and ER-negative metastatic lymph nodes?P=0.0092?.11.Data mining in GEO database,TCGA database and Target scan database showed that microRNA cluster c19mc had regulatory effect on ERa and GATA3 in triple negative breast cancer?P<0.0001?.12.The MDA-MB-231 cell lines were transfected with microRNA519a inhibitor and NC,GATA3 plasmid and empty PCMV6,and detected by RT-PCR and Western blot.It can be seen that the expression of GATA3 and ERalpha increased after the transfection of microRNA-519a inhibitor with statistical significance?P<0.01?.13.GATA3 plasmid was transfected into triple-negative breast cancer cell line MDA-MB-231 for culture.Western blot analysis showed that there was very little ER alpha expression in MDA-MB-231 cells compared with empty cells.Conclusion:1.The differential expression of ER and PR in lymph nodes and primary lesions has no significant effect on DFS and OS in patients with TNBC.2.In breast cancer patients with differentially expressed primary lesions and metastatic lymph nodes,there were significant differences in DFS and OS between groups.The positive expression group had a higher risk of recurrence/metastasis or death and a shorter survival time.3.The number of axillary lymph node metastases and total lymph nodes were correlated with DFS in TNBC patients.4.The number of axillary lymph node metastases and positive expression in lymph nodes were correlated with OS in TNBC patients.1.The expression of GATA3 in ER-positive metastatic lymph nodes was higher than that in ER-negative metastatic lymph nodes and primary lesions.2.The expression level of microRNA-519a in ER-positive metastatic lymph nodes was significantly lower than that in primary and ER-negative metastatic lymph nodes?P=0.0092?.3.The expression level of microRNA519a in tissues was negatively correlated with GATA3 and ERalpha.4.Data mining and prediction found that the expression level of c19mc of microRNAs was negatively correlated with GATA3 and ERERalpha.5.After the transfection of microRNA-519a inhibitor and NC into triple-negative breast cancer cell line MDA-MB-231,it can be seen that the expression of GATA3 and ERalpha in cells changed significantly after inhibiting the expression of microRNA-519a,but Western blot indicated that the protein levels of GATA3 and ERalpha did not change significantly.Considering that only microRNA-519a is not enough to support the whole process of ER conversion.6.After transfection of GATA3,ERA changed from Yin to yang,but weakly.It is not directly proportional to the level of GATA3,which indicates that there are other factors involved in the regulation of ER expression by GATA3.7.c19mc is involved in the regulation of GATA3 expression in upstream ER and ER expression,which affects the final expression level of ER.