Prognostic Marker In Breast Cancer Research Articles

CHEMOTHERAPY INDUCED HISTOPATHOLOGICAL CHANGES IN MASTECTOMY SPECIMENS - STUDY AT A TERTIARY CARE CENTRE

Background: Pathological complete response (pCR) to Neo adjuvant chemotherapy (NACT) is a signicant prognostic marker in breast cancer, so accurate diagnosis and grading of pathological chemo response of tumour is needed for an effective planning of treatment regimen. To study the various histopathological Aims: changes in Mastectomy specimens following NACT and to evaluate the pathological response of tumour to NACT, also to compare the expression of ER, PR, and HER2 in biopsies and post NACT mastectomy specimens by Immunohistochemistry (IHC). Methods And Materials: 50 breast carcinoma cases, who was diagnosed on trucut biopsies and treated with preoperative NACT followed by mastectomy was included. In post NACT mastectomy specimens, the chemoresponse was classied according to Union for International Cancer Control (UICC) guidelines. ER, PR, and HER2/ neu receptor expression was compared between trucut biopsy specimens (pre- NACT) and resection specimens (post- NACT). Results were analysed by Chi square test and McNemar chi square test. Among 50 cases, patients presented with lump, Results: involved more on the right and in upper outer quadrant. Most common stage at presentation is stage III A. Histologically all belong to IDCC -NST histological subtype. Signicant reduction in mean tumour size by 52.7% after NACT was observed. pCR, pPR and pNR was observed in 20 %, 58 % and 22 %of the cases respectively. Most common histological change seen in post NACT is necrosis, desmoplasia, brosis, lymphocytic inammatory response. Discordance in ER expression (17.5 %), PR expression (20%) and HER2 /neu (12.5 %) before and after NACT. But these were statistically insignicant. CONCLUSIONS: Pathological evaluation of chemo response of tumour is the gold standard. The discordance rate of ER, PR and HER-2/neu expression following NACT was statistically insignicant, larger studies needed to analyse their impact in patient management.

Identifying enhancer-driven subtype-specific prognostic markers in breast cancer based on multi-omics data.

Breast cancer is a cancer of high complexity and heterogeneity, with differences in prognosis and survival among patients of different subtypes. Copy number variations (CNVs) within enhancers are crucial drivers of tumorigenesis by influencing expression of their targets. In this study, we performed an integrative approach to identify CNA-driven enhancers and their effect on expression of target genes in four breast cancer subtypes by integrating expression data, copy number data and H3K27ac data. We identified 672, 555, 531, 361 CNA-driven enhancer-gene pairs and 280, 189, 113 and 98 CNA-driven enhancer-lncRNA pairs in the Basal-like, Her2, LumA and LumB subtypes, respectively. We then reconstructed a CNV-driven enhancer-lncRNA-mRNA regulatory network in each subtype. Functional analysis showed CNA-driven enhancers play an important role in the progression of breast cancer subtypes by influencing P53 signaling pathway, PPAR signaling pathway, systemic lupus erythematosus and MAPK signaling pathway in the Basal-like, Her2, LumA and LumB subtypes, respectively. We characterized the potentially prognostic value of target genes of CNV-driven enhancer and lncRNA-mRNA pairs in the subtype-specific network. We identified MUM1 and AC016876.1 as prognostic biomarkers in LumA and Basal-like subtypes, respectively. Higher expression of MUM1 with an amplified enhancer exhibited poorer prognosis in LumA patients. Lower expression of AC016876.1 with a deleted enhancer exhibited poorer survival outcomes of Basal-like patients. We also identified enhancer-related lncRNA-mRNA pairs as prognostic biomarkers, including AC012313.2-MUM1 in the LumA, AC026471.4-PLK5 in the LumB, AC027307.2-OAZ1 in the Basal-like and AC022431.1-HCN2 in the Her2 subtypes. Finally, our results highlighted target genes of CNA-driven enhancers and enhancer-related lncRNA-mRNA pairs could act as prognostic markers and potential therapeutic targets in breast cancer subtypes.

Role of epigallocatechin-3- gallate in the regulation of known and novel microRNAs in breast carcinoma cells.

Breast cancer comprises 30% of all cancer cases among the world’s women population. MicroRNAs are small, endogenous, non-coding RNAs that regulate cell proliferating and apoptotic pathways by modulating expressions of related genes. Phytochemicals like epigallocatechin-3-gallate (EGCG) are known to have a chemotherapeutic effect on cancer often through the regulation of microRNAs. The aim is to find out the key known and novel miRNAs, which are controlled by EGCG in breast cancer cell line MDA-MB-231. Next-generation sequencing (NGS) revealed 1,258 known and 330 novel miRNAs from untreated and 83 μM EGCG (IC50 value of EGCG) treated cells. EGCG modulated 873 known and 47 novel miRNAs in the control vs. treated sample. The hypothesis of EGCG being a great modulator of miRNAs that significantly control important cancer-causing pathways has been established by analyzing with Kyoto Encyclopedia of Genes and Genomes (KEGG) and Protein Analysis Through Evolutionary Relationships (PANTHER) database. Validation of known and novel miRNA expression differences in untreated vs. treated cells was done using qPCR. From this study, a few notable miRNAs were distinguished that can be used as diagnostics as well as prognostic markers for breast cancer.

Absolute lymphocyte count decreases with disease progression and is a potential prognostic marker for metastatic breast cancer.

Peripheral blood parameters such as the neutrophil-to-lymphocyte ratio (NLR) are prognostic markers for breast cancer patients. For instance, patients with a high NLR have a poor prognosis. Meanwhile, high absolute lymphocyte count (ALC) is reportedly a predictive factor for some chemotherapies. However, the underlying mechanisms behind how these markers relate to patient outcomes and how these markers change during the clinical course of patients with metastatic breast cancer (MBC) remains unknown. We retrospectively investigated 156 patients who were treated for MBC and eventually transitioned to best supportive care (BSC) at our hospital between January 2017 and December 2021. Changes in peripheral blood parameters during MBC treatments and their association with patient outcomes were examined. From the time of MBC diagnosis (baseline) through to the transition to BSC, ALC became significantly lower, while the NLR and platelet-to-lymphocyte ratio (PLR) became significantly higher (p < 0.001 for all). This association was independent of hormone receptor status. Cox proportional hazard modeling found patients with hormone receptor-negative and a lower baseline ALC had a significantly shorter overall survival (p = 0.030 and p = 0.019, respectively). We observed that peripheral blood markers gradually changed with MBC disease progression. Our data suggest that baseline ALC may be a potential prognostic marker after recurrence.

Role of mitochondrial translation in remodeling of energy metabolism in ER/PR(+) breast cancer.

Remodeling of mitochondrial energy metabolism is essential for the survival of tumor cells in limited nutrient availability and hypoxic conditions. Defects in oxidative phosphorylation (OXPHOS) and mitochondrial biogenesis also cause a switch in energy metabolism from oxidative to aerobic glycolysis contributing to the tumor heterogeneity in cancer. Specifically, the aberrant expressions of mitochondrial translation components such as ribosomal proteins (MRPs) and translation factors have been increasingly associated with many different cancers including breast cancer. The mitochondrial translation is responsible for the synthesis 13 of mitochondrial-encoded OXPHOS subunits of complexes. In this study, we investigated the contribution of mitochondrial translation in the remodeling of oxidative energy metabolism through altered expression of OXPHOS subunits in 26 ER/PR(+) breast tumors. We observed a significant correlation between the changes in the expression of mitochondrial translation-related proteins and OXPHOS subunits in the majority of the ER/PR(+) breast tumors and breast cancer cell lines. The reduced expression of OXPHOS and mitochondrial translation components also correlated well with the changes in epithelial-mesenchymal transition (EMT) markers, E-cadherin (CHD1), and vimentin (VIM) in the ER/PR(+) tumor biopsies. Data mining analysis of the Clinical Proteomic Tumor Analysis Consortium (CPTAC) breast cancer proteome further supported the correlation between the reduced OXPHOS subunit expression and increased EMT and metastatic marker expression in the majority of the ER/PR(+) tumors. Therefore, understanding the role of MRPs in the remodeling of energy metabolism will be essential in the characterization of heterogeneity at the molecular level and serve as diagnostic and prognostic markers in breast cancer.

Knockdown of CKAP2 Inhibits Proliferation, Migration, and Aggregate Formation in Aggressive Breast Cancer.

Simple SummaryCancer is a complex disease where cells grow and divide in an uncontrolled manner. It is well established that its development and progression involve major alterations in the activity of mitotic regulators. In order to improve our understanding of the contribution of cell-cycle progression defects to the development of disease, the aim of this study is to identify genes relevant to the proper progression of mitosis that are deregulated in breast cancer. Our findings identified CKAP2 as an important mitotic regulator in BC tumors. Moreover, in vitro experiments showed that gene silencing of CKAP2 blocked cell growth, cell migration, and formation of cell aggregates. These results demonstrated the important role of CKAP2 in breast cancer tumor formation.Loss of mitotic regulation is commonly observed in cancer and is a major cause of whole-chromosome aneuploidy. The identification of genes that play a role in the proper progression of mitosis can help us to understand the development and evolution of this disease. Here, we generated a list of proteins implicated in mitosis that we used to probe a patient-derived breast cancer (BC) continuum gene-expression dataset generated by our group by human transcriptome analysis of breast lesions of varying aggressiveness (from normal to invasive). We identified cytoskeleton-associated protein 2 (CKAP2) as an important mitotic regulator in invasive BC. The results showed that CKAP2 is overexpressed in invasive BC tumors when compared with normal tissues, and highly expressed in all BC subtypes. Higher expression of CKAP2 is also related to a worse prognosis in overall survival and relapse-free survival in estrogen receptor (ER)-positive and human epidermal growth factor receptor type 2 (HER2)-negative BC patients. Knockdown of CKAP2 in SKBR3 cells impaired cell proliferation and cell migration and reduced aggregate formation in a 3D culture. Our results show the important role of CKAP2 in BC tumorigenesis, and its potential utility as a prognostic marker in BC.

CCDC69 is a prognostic marker of breast cancer and correlates with tumor immune cell infiltration.

PurposeBreast cancer (BC) is the most common malignancy and the leading cause of cancer-related death among women worldwide. Early detection, treatment, and metastasis monitoring are very important for the prognosis of BC patients. Therefore, effective biomarkers need to be explored to help monitor the prognosis of BC patients and guide treatment decisions.MethodsIn this study, the relationship between CCDC69 expression levels and tumor clinical characteristics were analyzed using RNA-seq information in BC samples from the TCGA database. Kaplan-Meier survival analysis was performed to analyze the prognostic value of CCDC69 in BC patients. Besides, gene enrichment analysis in BC samples was used to confirm the main function of CCDC69 in BC. The correlation between the expression of CCDC69 and the number of tumor-infiltrating lymphocytes was confirmed by interaction analysis of TIMER and GEPIA.ResultsThe results showed that CCDC69 expression was significantly lower in cancer samples than in normal tissues, and was significantly lower in highly invasive BC than in carcinoma in situ. Meanwhile, low levels of CCDC69 were associated with a further poor prognosis. CDCC69 expression was positively correlated with the amount of different tumor-infiltrating lymphocytes. Mechanically, it could be presumed that the low expression of CCDC69 in BC might be caused by hypermethylation of the promoter region.ConclusionsSummarily, CDCC69 could be used as a potential biomarker to predict the prognosis of BC and the sensitivity to immunotherapy such as PD-1/PD-L1 checkpoint inhibitors.

Identification of UCP1 and UCP2 as Potential Prognostic Markers in Breast Cancer: A Study Based on Immunohistochemical Analysis and Bioinformatics.

Background: Uncoupling protein 1 (UCP1) and UCP2 are associated with tumor metabolism and immunity. However, the prognostic value and molecular mechanisms underlying their action in breast cancer (BC) remain unclear. Materials and methods: In TCGA-BRCA cohort, we investigated the expression characteristics of UCP mRNAs, analyzed their prognostic value by Kaplan-Meier survival analysis, their potential molecular functions by gene set enrichment analysis, and their relationship with immune infiltrating cell types using TIMER and CIBERSORT, along with the assessment of their association with mutational profiles. Kaplan-Meier survival analysis was performed for UCPs in our cohort and their association with BC thermogenesis was assessed by thermal tomography. Results: High expression of UCP1 and UCP2 were positive prognostic markers for BC. UCP1 was associated with the impaired glucose metabolism, while UCP2 with enhanced anti-tumor immunity. High expressions of UCP1 and UCP2 were associated with CDH1 mutations. High UCP1 expression was associated with a high rate of thermogenesis in BC. Conclusions: These results implied a key role of UCP1 and UCP2 in prognosis, metabolism, and immune infiltration in BC. Further investigation of the relevant molecular mechanisms may provide new strategies for BC treatment.

Correlation of 18-fluorodeoxyglucose uptake values on PET-CT scan with histological prognostic markers in breast cancer.

Positron emission tomography-computed tomography (PET-CT) scan utilizes 18-fluorodeoxyglucose (18-FDG), based on the principle of higher glycolytic activity and reduced glucose-6-phosphatase levels in cancer cells. This imaging modality is usually advised in the metastatic evaluation of stage III breast cancer patients. The correlation of maximum standard uptake values of primary lesion with different pathological and molecular markers has not been studied extensively. Retrospective analysis of the data was performed from our prospectively maintained breast cancer database. All the patients who had undergone 18-FDG PET-CT scan at initial evaluation for staging between June 2017 and April 2020 were included in the study. One-way ANOVA test or Student's t-test as appropriate was performed to assess the difference of means in maximum standard uptake values (SUVmax) of the primary lesion and axillary nodes with clinical stage, histological grade, molecular subtype. Bonferroni post hoc test was also applied. Out of 388 patients in the breast cancer database, 45 patients met inclusion criteria. There was a significant correlation of molecular subtype (p=0.029) with SUVmax of the primary lesion. Higher primary SUVmax was associated with higher T stage (p=0.01) and higher histological grade (p=0.06). In each molecular subtype, there was an increase in mean SUVmax of the primary lesion with increasing histological grade and vice versa. SUVmax of the primary lesion in breast cancer patients reflects tumor biology. Higher SUVmax can predict patients with triple-negative breast cancers and higher grades in primary tumors. However, further large-scale validatory studies are essential.

Dissecting the Roles of PDCD4 in Breast Cancer.

The human programmed cell death 4 (PDCD4) gene was mapped at chromosome 10q24 and encodes the PDCD4 protein comprised of 469 amino acids. PDCD4 inhibits protein translation PDCD4 inhibits protein translation to suppress tumor progression, and its expression is frequently decreased in breast cancer. PDCD4 blocks translation initiation complex by binding eIF4A via MA-3 domains or by directly binding 5’ mRNA internal ribosome entry sites with an RNA binding domain to suppress breast cancer progression and proliferation. Numerous regulators and biological processes including non-coding RNAs, proteasomes, estrogen, natural compounds and inflammation control PDCD4 expression in breast cancer. Loss of PDCD4 expression is also responsible for drug resistance in breast cancer. HER2 activation downregulates PDCD4 expression by activating MAPK, AKT, and miR-21 in aromatase inhibitor-resistant breast cancer cells. Moreover, modulating the microRNA/PDCD4 axis maybe an effective strategy for overcoming chemoresistance in breast cancer. Down-regulation of PDCD4 is significantly associated with short overall survival of patients, which suggests that PDCD4 may be an independent prognostic marker for breast cancer.

Integrated analysis of pivotal biomarker of LSM1, immune cell infiltration and therapeutic drugs in breast cancer.

The discovery of early diagnosis and prognostic markers for breast cancer can significantly improve survival and reduce mortality. LSM1 is known to be involved in the general process of mRNA degradation in complexes containing LSm subunits, but the molecular and biological functions in breast cancer remain unclear. Here, the expression of LSM1 mRNA in breast cancer was estimated using The Cancer Genome Atlas (TCGA), Oncomine, TIMER and bc‐GenExMiner databases. We found that functional LSM1 inactivation caused by mutations and profound deletions predicted poor prognosis in breast cancer (BRCA) patients. LSM1 was highly expressed in both BRCA tissues and cells compared to normal breast tissues/cells. High LSM1 expression is associated with poorer overall survival and disease‐free survival. The association between LSM1 and immune infiltration of breast cancer was assessed by TIMER and CIBERSORT algorithms. LSM1 showed a strong correlation with various immune marker sets. Most importantly, pharmacogenetic analysis of BRCA cell lines revealed that LSM1 inactivation was associated with increased sensitivity to refametinib and trametinib. However, both drugs could mimic the effects of LSM1 inhibition and their drug sensitivity was associated with MEK molecules. Therefore, we investigated the clinical application of LSM1 to provide a basis for sensitive diagnosis, prognosis and targeted treatment of breast cancer.

Integrated analysis of pivotal biomarker, immune cell infiltration, and therapeutic drugs for LSM1-mutated in breast cancer.

e13011 Background: The discovery of early diagnosis and prognostic markers for breast cancer can significantly improve survival and reduce mortality. LSM1 is known to be involved in the general process of mRNA degradation in complexes containing LSm subunits, but the molecular and biological functions in breast cancer remain unclear. Methods: In the present study, we investigated the expression profiles, prognostic roles, and genetic alterations of LSM1 in patients with breast cancer through several public databases, containing Oncomine and Gene Expression Profiling Interactive Analysis, Human Protein Atlas, Kaplan–Meier plotter and cBioPortal. Then, we constructed the protein-protein interaction networks of LSM1 proteins and their interactors by using the String database and Cytoscape software. In addition, we performed the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichmente. Finally, we explored the mechanisms underlying LSM1 involvement in BRCA by using gene set enrichment analysis. Results: We found that functional LSM1 inactivation caused by mutations and profound deletions predicted poor prognosis in BRCA patients. LSM1 was highly expressed in both BRCA tissues and cells compared to normal breast tissues/cells. High LSM1 expression is associated with poorer overall survival and disease-free survival. The association between LSM1 and immune infiltration of breast cancer was assessed by TIMER and CIBERSORT algorithms. LSM1 showed a strong correlation with various immune marker sets. Most importantly, pharmacogenetic analysis of BRCA cell lines revealed that LSM1 inactivation was associated with increased sensitivity to refametinib, and trametinib. However, both drugs could mimic the effects of LSM1 inhibition and their drug sensitivity was associated with MEK molecules. Therefore, we investigated the clinical application of LSM1 to provide a basis for sensitive diagnosis, prognosis and targeted treatment of breast cancer. Conclusions: In brief, our finding may contribute to increasing currently limited prognostic biomarkers and treatment options for breast cancer.

Low CRIM1 Levels Predict Poor Prognosis in Breast Cancer Patients.

BackgroundCRIM1 is involved in the development and preservation of the nervous system, capillary development, and vascular maintenance. Although CRIM1 was reported to involve in multiple cancers, its role in breast cancer is unclear.MethodsWe investigated CRIM1 expression levels using Oncomine, HPA, and immunohistochemistry analyses. BC-GenExMiner was employed to evaluate the relationship of CRIM1 expression with the clinicopathological characteristics of breast cancer. Its association with breast cancer prognosis was assessed by Kaplan-Meier analysis and PrognoScan. The correlation of the expression of CRIM1 with tumor immune infiltration was explored via TIMER. Gene set enrichment analysis (GSEA) was utilized to determine the cascades that are linked to CRIM1 in breast cancer. Finally, we explored CRIM1 and its co-expressed genes using R (3.6.3).ResultsHere, we find that CRIM1 expression was downregulated in various subtypes of breast cancer, and it was lowest in triple-negative breast cancers. ER and PR status were positively correlated with CRIM1 expression, while HER-2 expression was negatively correlated with CRIM1 expression. But in our immunohistochemical results in breast cancer specimens collected from our laboratory, HER-2 expression was positively correlated with CRIM1 expression. The expression of CRIM1 was correlated with menopause status, T stage, pathologic stage, histological type, and P53 status but not with age, N-stage, M-stage, Radiation therapy, and BRCA1/2 status. Survival analysis found that low CRIM1 expression was correlated with poorer DMFS, RFS and OS. Notably, CRIM1 expression was positively linked to the level of infiltration by CD8+ T-cells, endothelial cells, and neutrophils, and negatively linked to NK, B-cells, CD4+ T-cells, tumor purity, macrophage M1, and Tregs. Besides, DIXDC1 and PFDN6 were correlated to CRIM1 possibly.ConclusionOur findings demonstrated that low CRIM1 expression predict poor prognosis of breast cancer and CRIM1 might be used as a possible treatment target or prognostic marker in breast cancer. More researches are needed to better understand the prognostic value of CRIM1 in breast cancer.

Toll Like Receptor 4 Inhibition Improves Antitumor Responses to MBQ‐167 in Triple Negative Breast Cancer Cells in vitro

Triple negative breast cancer (TNBC) is an aggressive variant of breast cancer that lacks the expression of estrogen, progesterone, and human epithelial growth factor receptors. Due to the scarce availability of receptor‐targeted or hormonal treatments, TNBC patients are submitted to taxane‐based chemotherapies like paclitaxel. Patients initially respond to paclitaxel, but 4 out of 10 patients may develop tumor recurrence along with high metastasis and mortality rates. Several studies demonstrate that paclitaxel may induce chemoresistance and metastasis through the activation of Toll‐Like Receptor 4 (TLR4)/NFKB signaling pathway, a known cancer prognostic marker in breast cancer. Hence, new treatments are needed to improve TNBC prognosis in patients. Recently, our laboratory and collaborators developed MBQ‐167, a potent Rac and Cdc42 inhibitor that decreases tumor burden, cell proliferation, and metastasis in TNBC mouse models. In preliminary studies with TNBC mice, we observed that MBQ‐167/Paclitaxel combination treatment prevented metastasis progression compared to paclitaxel treatment alone. This led us to hypothesize that MBQ‐167 chemosensitizes aggressive breast cancer cells and reduces metastasis by blocking Rac/Cdc4, hence decreasing their downstream target NfkB activity. This research project aims to evaluate cellular and molecular events that explain how MBQ‐167 prevents paclitaxel‐induced metastasis. Here, we generated TLR4‐knockdowns (TLR4‐KD) in MDA‐MB‐231 TNBC cell lines using lentiviral particles with shRNAs against the TLR4 mRNA. After knockdown validation with western blot analysis (WB), Wildtype (MDA‐MB‐231), Scramble control knockdowns (Sc‐KD), and TLR4‐KD cells were treated with different concentrations of MBQ‐167, paclitaxel, doxorubicin, or combinations. After treatments, we evaluated the effect on cell viability from MTT assays at 96 and 120 hours, cell apoptosis through Caspase‐Glo 3/7 Assays at 48 hours, and cellular migration through scratch wound healing assays. Our results demonstrate that partial inhibition of TLR4 decreases the cell viability of cells, increases apoptosis, and decreases cell migration after treatment with MBQ‐167 (250 nM and 500 nM), Paclitaxel (5 nM and 10 nM), or Doxorubicin (250 nM and 500 nM). We also observed an additive response upon combinatorial treatments with MBQ‐167/Paclitaxel and MBQ‐167/Doxorubicin when TLR4‐KD cells were compared to wild‐type and Sc‐KD cells. Altogether, these results suggest that TLR4‐KD chemosensitizes cells to Paclitaxel and Doxorubicin chemotherapies. In addition, TLR4 inhibition improves cellular response to Rac/Cdc42 inhibitor, MBQ‐167. Therefore, TLR4‐KD may reduce Rac and NfkB signaling, improving treatment response to MBQ‐167, MBQ‐167/Paclitaxel, and MBQ‐167/Doxorubicin. Other cellular studies, including Rac and NfkB activation assays, are being evaluated in our laboratory.

TGF-β1 Promotes Autophagy and Inhibits Apoptosis in Breast Cancer by Targeting TP63.

BackgroundBreast cancer (BC) is a prevalent female cancer, which has high morbidity and mortality. However, the pathogenesis of BC has not been fully elucidated. Studies have shown that TGF-β1 plays an important role in regulating the balance between autophagy and apoptosis of tumor. We aim to clarify the specific mechanism of autophagy and apoptosis in breast cancer maintaining the tumor microenvironment.MethodsThe clinical characteristics of 850 BC patients were retrieved from the TCGA database. Differentially expressed autophagy-related genes (DEARGs) between tumor and normal tissues were obtained by the Wilcox test. Through Cox proportional hazard regression analysis, the prognostic risk model was constructed and verified by the ROC curve. We used MDC staining, colony formation assay, CCK-8, flow cytometric analysis to confirm the importance of TGF-β1 on the autophagy and apoptosis of breast cancer cells. Furthermore, western blot was performed to determine the relative expression of protein. The Kaplan-Meier Plotter database was utilized to identify the prognostic value of TP63.ResultsWe successfully constructed a prognostic risk model of breast cancer and screened out an autophagy-related prognostic gene -TP63. We predicted that TGF-β1 and TP63 have a binding site in the JASPAR database as expected. Additionally, TGF-β1 promoted autophagy and inhibited apoptosis of breast cancer cells by inhibiting the expression of TP63.ConclusionOur study demonstrated that the molecular mechanism of TGF-β/TP63 signaling in regulating autophagy and apoptosis of breast cancer and provided a potential prognostic marker in breast cancer.

Promotion of tumorigenesis by miR‐1260b–targeting CASP8: Potential diagnostic and prognostic marker for breast cancer

MicroRNAs are reported as promising biomarkers for the diagnosis and treatment of breast cancer. miR‐1260b is identified as a tumor‐associated noncoding microRNA in other cancers, although the role of miR‐1260b and its clinical relevance in breast cancer remain unclear. In this study, miR‐1260b as a potential prognostic biomarker was observed by univariate and multivariate Cox regression analyses in 102 breast tumor tissues. The tumorigenic role of miR‐1260b in terms of proliferation, apoptosis, and migration of breast cancer cells was investigated using gain‐ and loss‐of‐function assays in vitro. Additionally, the potential early diagnosis and treatment monitoring marker of miR‐1260b was validated in 129 plasma samples. We found that high miR‐1260b expression was markedly associated with bulky tumor size, advanced stage, and lymph node invasion. Particularly, the high–miR‐1260b‐expression group showed shorter overall survival than the low–miR‐1260b‐expression group. The inhibition of oncogenic miR‐1260b induced apoptosis and decreased migration and invasion of MDA‐MB‐231 cells. CASP8 was revealed as a direct target gene of miR‐1260b, which is closely related to apoptosis. Furthermore, miR‐1260b expression levels in plasma were significantly higher in patients with breast cancer than in healthy controls. The patients who tested positive for miR‐1260b showed 16.3‐ and 18.2‐fold higher risks in the early stage and locally advanced stage, respectively, compared with healthy controls, and the risk was decreased 6.2‐fold after neoadjuvant chemotherapy. Taken together, miR‐1260b may be a potential novel diagnostic, prognostic, and therapeutic target in breast cancer.

Abstract P4-05-07: Assistance with an artificial intelligence-powered tumor infiltrating lymphocytes (TIL) analyzer reduces interobserver variation in pathologic scoring of TIL in breast cancer

Abstract Background Tumor infiltrating lymphocytes (TIL) is a promising prognostic marker in breast cancer. However, TIL is manually scored by pathologists, thus laborious work is required and interobserver heterogeneity exists in the results. In this study, we aimed to evaluate the clinical utility of an artificial intelligence (AI)-powered TIL analyzer in terms of reducing the interobserver variation. Methods Lunit SCOPE IO, AI-powered TIL analyzer was trained and validated with a 2.8 x 109 micrometer2 area and 5.9 x 106 TIL from 3,166 H&amp;E Whole-Slide Images (WSI) of multiple cancer types including breast cancer, annotated by 52 board-certified pathologists. Three independent board-certified pathologists scored TIL% of H&amp;E slides of breast cancer from an external cohort (N = 199). TIL% was calculated referenced on the guideline of Immuno-Oncology Biomarker Working Group on Breast Cancer. For the cases of TIL score difference between each pathologist and AI model more than 15%, the pathologists were asked to revise TIL% in assistance with AI model which displays both stromal area and TIL. Finally, we compared the interobserver variation based on intraclass correlation coefficients (ICC) before and after AI assistance. Results The distribution of TIL score by 3 pathologists was 7% (5-20%), 15% (5-50%), and 20% (10-40%), respectively [median (25%-75% quantile)]. The ICC value of the initial TIL score evaluation was 0.716 (95% confidence interval, 0.560-0.811). Afterward, pathologists revised their initial scoring with assistance of AI model for the cases of difference more than 15% (n = 19, 72, and 73, respectively for each pathologist). After rescoring, number of slides with 15% or more difference of TIL% between raters significantly decreased from 109 slides (54.8%) to 75 slides (37.7%, p &amp;lt; 0.001). The ICC value after re-scoring TIL% was 0.831 (95% confidence interval, 0.725-0.890). Conclusions There was a notable interobserver variation to score TIL% in breast cancer. Assistance with AI-powered TIL analyzer substantially improved the pathologist’s consensus and could be regarded as one of references for the final labeling of TIL%. Citation Format: Soo Ick Cho, Wonkyung Jung, Sangjoon Choi, Seokhwi Kim, Sanghoon Song, Gahee Park, Minuk Ma, Seonwook Park, Sergio Pereira, Sangheon Ahn, Brian Jaehong Aum, Seunghwan Shin, Kyunghyun Paeng, Donggeun Yoo, Chan-Young Ock. Assistance with an artificial intelligence-powered tumor infiltrating lymphocytes (TIL) analyzer reduces interobserver variation in pathologic scoring of TIL in breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-05-07.

SERUM IRON PARAMETERS AS PROGNOSTIC MARKERS IN BREAST CANCER PATIENTS

BACKGROUND AND OBJECTIVE: Breasts are an important aspect of a woman's femininity and all aberrations in their functional and cosmetic attributes are essential for scientic evaluation as breast cancer has become the most common cancer among women worldwide. Its risk increases with age and menopausal status. Serum parameters like Ferritin and TIBC concentration is altered in sera of these patients. Iron homeostasis is inuenced by estrogen and overload impacts cellular proliferation and physiological dysfunction in electron and oxygen transport, energy production and DNA synthesis. A vicious cycle between breast cancer, iron homeostasis deregulation, menopausal status and serum parameters derangement enables us to prognosticate such patients. METHOD: Histopathologically conrmed, 50 newly diagnosed cases were analyzed with age matched 50 clinically healthy controls with no family history. Level of serum ferritin was estimated by Sandwich Elisa using Ferritin SAElisa kit and serum TIBC level measured with CL-1000i Chemiluminescence immunoassay analyzer. RESULT: Serum ferritin level in breast cancer cases (300.73±25.33ng/ml) was statistically higher than in controls (85.22±41.80ng/ml). In breast cancer, ferritin level was higher in postmenopausal (300.73±25.33ng/ml) compared to premenopausal (228.059±11.24ng/ml) patients and even in their healthy counterparts.Serum TIBC level in breast cancer (772.99±127.93) was higher than in controls (329.41± 69.40).In postmenopausal breast cancer female TIBC level was higher (815.39±104.32) compared to premenopausal females (652.31±110.86) and similarly it was higher in postmenopausal healthy controls than in premenopausal controls and signicant statistically. CONCLUSION: Serum ferritin and TIBC parameters can be used as prognostic markers for breast cancer and their levels are elevated in postmenopausal females of both breast cancer patients and healthy cases.

Common Sensitive Prognostic Marker in Breast Cancer and their Clinical Significance: A Review Article

The most common cancer in women is breast cancer. Also, it is the biggest cause of death in women with cancer. The exact cause is not known but there are risk factors such as women above 55, Caucasian races, late age of first pregnancy, genetic factors and, family history. It has two classifications histological and molecular. Tumor biomarkers are either prognostic markers or predictive markers. We here present a review for breast cancer prognostic markers. Breast cancer prognostic markers are d

Comparative Analysis of Ki-67 in Different Scoring Patterns and its Association with other Prognostic Markers of Breast Carcinoma

Introduction: Immunohistochemical evaluation of Ki-67 is widely used for estimation of tumour proliferation in breast cancer. Till date, no specific method or a cut-off point for Ki-67 exists. Aim: To perform a comparative analysis between different scoring patterns and mean Ki-67 value and association of mean Ki-67 with other prognostic markers like tumour size, lymph node status, tumour stage and grade and different molecular subtypes of breast cancer. Materials and Methods: A cross-sectional study was conducted at Dr. D.Y Patil Medical College, Hospital and Research Centre, Pune, Maharashtra, India, between August 2019 to August 2021. Total of 50 new diagnosed cases of breast cancer were studied for the histologic type, grade and stage of the tumour. Immunohistochemistry for Oestrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal Growth Factor Receptor 2 (HER2) and Ki-67 was performed. Association of Ki-67 with other prognostic markers like tumour size, lymph node status, tumour stage and grade and different molecular subtypes of breast cancer was evaluated by expressing Ki-67 as a continuous variable (mean±SD) and also by dividing Ki-67 into different scoring patterns (I: ≤14%, &gt;14%, II: ≤15%, 16- 30%, &gt;30% and III: &lt;20%, 20-50%, ≥50%). Statistical tests like Kruskal-Wallis test (mean Ki-67 with tumour size, tumour grade and molecular subtypes), Mann-Whitney Rank Sum Test (mean Ki-67 with lymph node status) and Analysis of variance (ANOVA) (mean Ki-67 with staging) respectively. Results: Out of 50 patients, 40 (80%) were older than 50-yearold. Twenty six (52%) cases affected the left breast. A total of 49 (98%) were diagnosed as Invasive Ductal Carcinoma (IDC). Among them 26 (52%) cases were of grade III and 25 (50%) cases were of Luminal A. Mean Ki-67 and molecular subtypes of breast cancer had statistically significant association (p= 0.002). No association was found between mean Ki-67 and tumour size (p=0.608), lymph node status (p=0.506) stage (p=0.979) and grade (p=0.095) of the tumour. Although scoring pattern I and III had no remarkable findings. Notably, scoring pattern II showed higher tumour sizes, lymph node positivity, higher stage and grade and basal-like tumours demonstrated a higher Ki-67 index. Conclusion: Evaluation of Ki-67 as a continuous variable yielded significant association with other prognostic markers of breast cancer. There was no single “best” scoring pattern identified. A direct association of Ki-67 was found with molecular subtypes of breast cancer.