| Objective:1.Detect the abundance of Pericarcinomatous tissue infiltrating lymphocytes and tumor infiltrating lymphocytes in adjacent breast cancer with different molecular subtypes;2.Detect the expression of negative co-stimulating factors in adjacent breast cancer Pericarcinomatous tissue infiltrating lymphocytes and tumor infiltrating lymphocytes;3.To explore the relationship between the expression of negative co-stimulating factors in the tumor microenvironment and different molecular subtypes,clinical stages and clinical pathological features of invasive breast cancer,and to analyze the clinical significance.Methods:1.According to the inclusion and exclusion criteria,51 patients admitted to the First Hospital of Shanxi Medical University for breast cancer modified radical resection from October 2020 to October 2021 were selected to collect tumor tissue and paracancer tissue(paracous tissue is defined as tissue within 2 cm from the edge of cancerous tissue);2.SPSS25.0 software was used to statistically analyze the clinical data of each case,mainly including patient age,menstrual conditions,histological grading,immunohistochemical indicators,tumor size,axillary lymph node metastasis,presence or absence of distant metastasis,tumor molecular typing,and clinical TNM staging;3.Application of flow cytometry to detect the abundance of CD4+and CD8+infiltrating T cell subsets in tumor tissues and paracancerous tissues of patients;4.By flow cytometry,the co-expression levels of negative co-stimulators CTLA-4,PD-1,TIM-3 and TIGHT in tumor tissues and paracancer tissues of patients were detected,and the relationship between T lymphocyte negative sympimulatory factor coexpression and clinical case data in the tumor microenvironment of different molecular subtypes of breast cancer was analyzed.Results:1.Breast cancer patients with high histological grade,tumor diameter≧2 cm,axillary lymph node metastasis≧4,and distant metastases,the percentage of T lymphocytes with mono-expression PD-1,CTLA-4,TIM-3,TIGHT and co-expressing PD-1+ CTLA-4+,TIM-3+AND TIM-3+TIGHT+T lymphocytes in tumor tissue invasive lymphocytes(TIL)and adjacent tissue invasive lymphocytes(PIL)was significantly increased(P<0.001);2.Triple negative breast cancer(TN)TIL and PIL monoexpression PD-1,CTLA-4,TIM-3,TIGHT and co-expression of PD-1+CTLA-4+,TIM-3+TIGHT+ T lymphocyte percentage increased significantly(P<0.001),hormone receptor positive/human epidermal growth factor negative breast cancer(HR+/ERBB2-)and human epidermal growth factor positive(ERBB+)breast cancer did not observe a significant increase in the percentage of positive lymphocytes(P>0.05);3.Compared with stage Ⅰ.stage Ⅰ,the percentage of positive lymphocytes in TIL and PIL in stage Ⅱ.stage Ⅱ,Ⅲ.and Ⅳ.patients was significantly increased and was statistically significant(P<0.05),but for stage Ⅱ and PHASE Ⅲ patients,although with the increase of clinical stage,TIL and PIL positive lymphocytes showed a certain upward trend,but there was no obvious statistical difference between the two(P>0.05).Therefore,for patients with breast cancer who are at the beginning of diagnosis,it is a new clinical choice to use immunotherapy as soon as possible;4.Compared with PIL,the percentage of PD-1+CTLA-4+and TIM-3+TIGHT+double-positive T lymphocytes in TIL was higher(P<0.05).Conclusion:1.Negative co-stimulating factors PD-1,CTLA-4,TIM-3,TIGHT monoexpression in breast cancer tissue infiltrating lymphocytes and the coexpression of PD-1+ CTLA-4+,TIM-3+TIGHT+are related to the patient’s histological grade,tumor diameter,number of lymph node metastases,and distant metastases;2.There are differences in the expression of negative co-stimulating factors in breast cancer tissues in different molecular typings.Among them,the single expression and co-expression of negative co-stimulating factors in three-negative breast cancer were significantly increased(P<0.001),and the negative co-stimulating factor of hormone receptor positive/human epidermal growth factor-negative breast cancer and human epidermal growth factor positive breast cancer were not significantly increased(P>0.05).Therefore,in the clinical diagnosis and treatment process,patients with triple-negative breast cancer are more likely to benefit from immunotherapy;3.There are differences in the expression of negative co-stimulating factors in breast cancer tissues in different clinical stages.The percentage of positive lymphocytes in TIL and PIL in stage Ⅳ patients was significantly increased(P<0.001),and the positive expression of stage Ⅱ and STAGE Ⅲ patients showed an increase trend,but there was no obvious statistical difference(P>0.05);4.Invasive lymphocytes in the tumor microenvironment can observe more pronounced failure than paracancer tissue,which may indicate that the closer to the tumor tissue,the gradual increase in the concentration of related cytokines that induce T lymphocyte failure,we can reactivate the immune surveillance function of T lymphocytes by further blocking the secretion of related cytokines. |
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