Clinicopathologic Research Of Ductal Carcinoma In Situ With Microinvasion

Purpose:1.To investigate the value of preoperative neutrophil/lymphocyte ratio（NLR）on predicting invasion in ductal carcinoma in situ and the prognosis of early breast cancer.2.To investigate the clinical,imaging,NLR,pathological characteristics,and prognosis of DCIS-MI and the difference with ductal carcinoma in situ（DCIS）and T1a-invasive ductal carcinoma（T1a-IDC）respectively,so that we could clarify the biological behavior of DCIS-MI and its stage in breast cancer’s development.We also sought to determine factors predicting microinvasion in DCIS to facilitate the doctors’clinical treatment strategies.3.To find new targets genes or its related proteins or miRNAs which could predict invasion.Method:1.338 cases of DCIS diagnosed by preoperative vacuum-assisted biopsy or rapid intraoperative frozen biopsy were identified and then divided into DCIS group（201）and IDC group（137）according to whether invasion was existed in postoperative pathology.NLR and clinicopathologic features were analyzed by Chi-square test or Mann-Whitney U testand Logistic regression for Univariate and multivariate analysis.ROC curve of NLR was made according to the dependent variable of invasion,AUC was applied to estimate the prediction performance of preoperative NLR.Then patients were reclassified as high-level NLR（H-NLR）（NLR>2.105）and low-level NLR（L-NLR）（NLR<2.105）.Chi-square test was used to compare clinical pathological characteristics between the two groups of patients.Survival analysis was performed by the Kaplan-Meier.P<0.05 was considered statistically significant.2.131 cases of DCIS,74 cases of DCIS-MIand73 cases of T1a-IDC were retrospectively analyzed using Chi-square test,or Mann-Whitney U test for Univariate analysis and Kaplan-Meier for survival analysis to compare DCIS-MI with DCIS and T1a-IDC in clinical performance,imaging,pathology and prognosis.The statistically significant factors in univariate analysis between DCIS and DCIS-MI were included into Logistic regression for multivariate analysis.The risk prediction model of DCIS with microinvasion was constructed.The risk threshold was established based on the ROC curve.Information of 43new cases of DCIS（35）and DCIS-MI（9）were included in this model to evaluate the accuracy of it.P<0.05 was considered statistically significant.3.Screen co-differentially expressed genes between DCIS and IDC based on the database Oncomine,bioinformatics softwares were used for GO enrichment analysis,KEGG Pathway analysis and PPI analysis to obtain key genes or proteins and predict their related miRNAs.Result:1.There was significant difference in preoperative NLR between DCIS group and IDC group.The AUC of preoperative NLR was 0.580,and the best cutoff value was 2.105.Univariate analysis showed that for patients diagnosed as DCIS preoperatively,NLR>2.105,tumor diameter>4.15cm,high nuclear grade and acne-like necrosis in the component of DCIS were predictors of invasion,and multivariate analysis confirmed NLR>2.105 as an independent predictor of invasion.The difference in tumor diameter,axillary lymph node status and family history between different NLR levels in early stage breast cancer were statistically significant.The differences of rest clinical pathological features,DFS and OS were not statistically significant between different NLR levels.2.The difference in tumor diameter,axillary lymph node status,morphology and distribution of calcification on mammography,distorted structure on mammography,nuclear grade and acne-like necrosis in component of DCIS,merely calcification and calcified mass on mammography between DCIS and DCIS-MI was statistically significant.Univariate and multivariate analysis showed that tumor diameter>4.15 cm,NLR>2.105,structural distortion,acne-like necrosisin DCIS components and merely calcification on mammography were statistical significance between the two groups.A microinvasion risk prediction model was constructed based on these with a probability of Pi=1/（1+e-Logit（P））,where Logit（P）==-1.169+1.233X₁（Tumor size）+0.724X₂（NLR level）+2.150X₃（acne-like necrosis）+2.212X₄（structural distortion）-1.088X₅（simply calcification）-0.525X₆（high nuclear grade）,the best cut-off value of Pi was 1.6357.The verification group tested the model for a specificity of 80%,a sensitivity of 86.9%and an accuracy of 81.8%.Compared with T1a-IDC,DCIS-MI had a higher positive rate of HER-2 and lower rate of mass performance in ultrasound,less blood flow,lower aspect ratio,while no significant differences in tumor size,axillary lymph node status,and immunohistochemistry.There was no statistical difference between DCIS-MI and DCIS and T1a-IDC in DFS and OS.3.A total of 52 co-differentially expressed genes were obtained.The GO enrichment,KEGG Pathway and PPI analysis showed these genes were mainly enriched in extracellular matrix-related pathways and functions and were closely related to the occurrence and development of invasion.Key genes was COL1A1and 9 related miRNAs were obtained.Conclusion:1.For patients with preoperative diagnosis of DCIS,preoperative NLR>2.105 may be an independent predictor of invasion.2.Tumors in patients with preoperative NLR>2.105 had a more aggressive biological performance,which may indicate poorer prognosis.3.Preoperative NLR>2.105,tumor diameter>4.15 cm,acne-like necrosis,structural distortion and simply calcification on mammography were independent impact factors of microinvasion.Risk prediction model based on these factors could help clinicians to effectively predict microinvasion.4.Compared with DCIS,DCIS-MI had a larger tumor size,higher axillary lymph node metastasis rate,more acne-like necrosis and higher grade,which made DCIS-MI know from DCIS as a kind of invasive carcinoma.DCIS-MI and T1a-IDC had some differences in morphology,but no significant differences was found in prognostic indicators.DCIS-MI could be considered as a subgroup of T1a-IDC.5.The differentially expressed genes between DCIS and IDC play an important role on breaking down of extracellular matrix and cell adhesion which were closely related to the occurrence of invasion.High expression of COL1A1or down-regulation of its related miRNAs might suggest DCIS with invasion even microinvasion.