1.Study Of Genes Related To Breast Cancer Axillary Lymph Node Metastasis Based On Whole Exome Sequencing 2.Clinicopathological And Prognostic Analysis Of Breast Intraductal Papillary Tumors

Background Breast cancer is the most common malignant tumor in women.Lymph node metastasis is the most important metastatic pathway of breast cancer.Among them,the axillary lymph node is the first station of regional lymph node metastasis.Axillary lymph node metastasis is an important factor affecting the prognosis of breast cancer.There are many factors that affect the axillary lymph node metastasis of breast cancer,but the mechanism of lymph node metastasis is still under exploration.Methods The study recruited ten patients of Chinese Han population with primary breast cancer and metastatic lymph nodes.Whole exome NGS was performed on DNA isolated from fresh-frozen tumor tissue,metastatic axillary lymph node and peripheral blood following the Illumina protocols.Single nucleotide variants(SNVs),insertion/deletions(Indels),and copy-number variants(CNVs)were generated.Comparison between tumor and lymph node provided an estimated of clonal relatedness within individual patients.Through a variety of bioinformatics methods to screen and associated with the genetic database of comparative analysis,we access to possible with the metastatic related mutations.And,the expression of protein in tumor cells,interstitial fibroblasts and metastatic lymph nodes was evaluated by immunohistochemical staining.Results The samples were sequenced to high,uniform coverage(average depth was 123×,with>99%of exons covered at greater than 100×).Over 40000 single nucleotide polymorphisms(SNPs)were detected in each sample.The average frequency of SNVs and InDels of primary tumor and metastatic axillary lymph node were 5066 vs 5141,1400 vs 1476,respectively.After filtering and cross matching,nine genes(ZNF697,AHNAK,TXNDC2,TCHH,UBQLNl,PCDHA8,ZNF254,QRICH2,ZNF888)were identified as significantly specific mutations in primary tumor and fifteen genes(TCHH,HRNR,ZAN,FRG1BP,FRG1DP,AHNAK,KRTAP4-6,AHNAK2,ZNF343,ZAN,RPTN,POM121,KRTAP10-4,GOLGA6L2,TAS2R30)were identified as significantly specific mutations in metastatic lymph node(False discovery rate[FDR]<0.1).We apply MutSigCV to exome sequences and discover extraordinary variation in mutation frequency and spectrum of HRNR and AHNAK2,which were overlapping genes in lymph node metastasis-specific genes.The average optical density of metastatic lymph nodes and normal lymph nodes were 0.0471±0.0185,0.0027±0.0006,respectively.T-test showed that the average optical density of metastatic lymph nodes was significantly higher than the normal lymph nodes(P=0.039).Conclusion The study provide evidence that primary tumor and metastatic lymph node revealed overlapping variants whose expression is associated with tumor invasion,and there were some mutations unique to the involved node,which may be suggested as genetic variability early in metastatic process.The results of different bioinformatics analysis showed that AHNAK2,HRNR and other gene mutations may be closely related to lymph node metastasis.Objective Intraductal papillary tumors are clinically common diseases derived from the ducts.The aim of the study is to investigate intraductal papillary tumors clinicopathological characteristics and influencing factors of canceration.Methods The clinicalpathological data of 674 patients with breast intraductal papillary tumors,who underwent surgery in National Cancer Center/Cancer hospital,Chinese academy of medical sciences from January 2010 to January 2016,were retrospective analyzed.Results The median follow up time was 46 months.The 674 patients were classified into intraductal papilloma(547 cases)and malignant intraductal papillary carcinoma(127 cases).The 3-year recurrence free survival rates in intraductal papilloma and atypical intraductal papilloma were 98.2%and 95%respectively.The log rank test showed a significant difference(P=0.009).Multivariate analysis indicated that the atypical ductal hyperplasia was a major prognostic factor affecting the recurrence free survival of intraductal papilloma(RR=0.183,95%CI=0.054?0.777,P=0.020).4 cases of intraductal papillary carcinoma patients had local recurrence.Logistic analysis results showed that patient aged older than 45,breast mass and maximum tumor diameter greater than 2cm were the possible clinical manifestations of malignant tumor(0R=1.735,95%CI=1.007?2.990,P=0.047;OR=2.849,95%CI=1.207?6.711,P?0.017;OR=3.792,95%CI=2.162?6.653,P<0.001).Conclusion Intraductal papillary tumors with a certain rate of recurrence.Age,clinical features and tumor size may be predictors of Intraductal papillary carcinoma.