Cases Of Breast Cancer Tissues Research Articles

DNAJA1 promotes proliferation and metastasis of breast cancer by activating mutant P53/NF-κB pathway

PurposeBreast cancer is one of the most common tumors with high malignancy and metastatic rate. DNAJA1 is closely related to tumor progress in several tumors. However, the role and mechanisms of DNAJA1 in the metastasis and proliferation of breast cancer are unknown. MethodsImmunohistochemistry and western blot were used to detect the protein expression genes. In vivo and vitro experiments were performed to evaluate the proliferation, invasive and metastatic abilities of breast cancer cells. ResultsDNAJA1 was high expressed in 234 cases of breast cancer tissues and associated with metastasis, p53 expression and poor survival for patients. Knock down of DNAJA1 decreased the number of plate clone formation and the OD value of CCK8 assays in breast cancer cells. Depletion of DNAJA1 also in decreased the invasive abilities of breast cancer cells. In vivo, knock down DNAJA1 decreased the growth of subcutaneous tumor and lung metastatic nodes. Mechanically, DNAJA1 could bind with P53-R175H and reduced its degradation. Up regulation of DNAJA1 in mutant P53-R175H breast cancer cell promoted the nuclear translocation of p65, activated NF-κB pathway and enhanced the transcription of its downstream genes such as MMP9, CXCL10 et al. Blockade of NF-κB pathway effectively rescued the effects of DNAJA1 on proliferation and metastasis in breast cancer. ConclusionOur study reveals that DNAJA1 is up regulated in breast cancer and promotes breast cancer cells proliferation and metastasis via P53-R175H/NF-κB pathway. It might be a potential prognosis marker for the breast cancer patients.

Overexpression of PBK/TOPK relates to poor prognosis of patients with breast cancer: a retrospective analysis

BackgroundPDZ-binding kinase/T-lymphokine-activated killer cell-derived protein kinase (PBK/TOPK) is a potential prognostic indicator for patients with breast cancer. The objective of the present study was to explore the relationship between PBK/TOPK expression and clinicopathological indicators as well as the survival of patients with breast cancer.MethodsImmunohistochemical staining was used to detect the expression of PBK/TOPK in 202 cases of breast cancer tissues. The relationship between PBK/TOPK and clinicopathological parameters was evaluated using Spearman’s rank-order correlation. The difference in PBK/TOPK expression among different molecular types was analyzed with the chi-square test. Kaplan-Meier analysis was used to create a survival curve and the log rank test was used to analyze the overall survival (OS) and disease-free survival (DFS). Prognostic correlation was assessed using univariate and multivariate Cox regression analyses.ResultsAmong 202 breast cancer samples, PBK/TOPK was expressed (“+” and “++”) in 182 samples (90.1%). In addition, the histological grade, TNM stages, lymph node metastasis, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), and Ki-67 were positively associated with PBK/TOPK expression. With regard to the molecular type, the expression of PBK/TOPK is different. The expression level of PBK/TOPK was negatively correlated with both the OS and DFS of breast cancer patients. The difference in the above results is meaningful (P < 0.05).ConclusionsPBK/TOPK is overexpressed in breast cancer, and the expression is closely related to the clinicopathological characteristics of the disease. Breast cancer patients with high expression of PBK/TOPK have a poor prognosis. Therefore, healthcare providers can optimize breast cancer management using this indicator.

Circular RNA circWWC3 augments breast cancer progression through promoting M2 macrophage polarization and tumor immune escape via regulating the expression and secretion of IL-4

Interaction between tumor cells and tumor microenvironment (TME) is critical to promote tumor progression and metastasis. As the most abundant immune cells in TME, macrophages can be polarized into M2-like tumor-associated macrophages (TAMs) which further promote tumor progression. However, to date, the molecular mechanisms of TAM polarization in TME are still largely unknown. In the present study, we revealed that circular RNA circWWC3 could up-regulate the expression and secretion of IL-4 in breast cancer cells. Enhanced secretion of IL-4 from breast cancer cells could augment the M2-like polarization of macrophages in TME, which further promotes the migration of breast cancer cells. In addition, increased secretion of IL-4 from breast cancer cells could induce the expression PD-L1 in M2 macrophages. Moreover, up-regulated IL-4 also enhanced the expression of PD-L1 in breast cancer cells, which further facilitates breast cancer immune evasion. Though analyzing the expression of circWWC3, IL-4, PD-L1, and CD163 in 140 cases of breast cancer tissues, we found that high expression of circWWC3 was associated with poor overall survival and disease-free survival of breast cancer patients. Breast cancer patients with circWWC3high/PD-L1high breast cancer cells and CD163high macrophages had a poorer overall survival and disease-free survival. Conclusively, circWWC3 might augment breast cancer progression through promoting M2 macrophage polarization and tumor immune escape via regulating the expression and secretion of IL-4. CircWWC3 might be a potential therapeutic target in breast cancer.

Silencing of KIF3B Suppresses Breast Cancer Progression by Regulating EMT and Wnt/β-Catenin Signaling.

Breast cancer is the most common malignant tumors in women. Kinesin family member 3B (KIF3B) is a critical regulator in mitotic progression. The objective of this study was to explore the expression, regulation, and mechanism of KIF3B in 103 cases of breast cancer tissues, 35 metastatic lymph nodes and breast cancer cell lines, including MDA-MB-231, MDA-MB-453, T47D, and MCF-7. The results showed that KIF3B expression was up-regulated in breast cancer tissues and cell lines, and the expression level was correlated with tumor recurrence and lymph node metastasis, while knockdown of KIF3B suppressed cell proliferation, migration, and invasion both in vivo and in vitro. In addition, UALCAN analysis showed that KIF3B expression in breast cancer is increased, and the high expression of KIF3B in breast cancer is associated with poor prognosis. Furthermore, we found that silencing of KIF3B decreased the expression of Dvl2, phospho-GSK-3β, total and nucleus β-catenin, then subsequent down-regulation of Wnt/β-catenin signaling target genes such as CyclinD1, C-myc, MMP-2, MMP-7 and MMP-9 in breast cancer cells. In addition, KIF3B depletion inhibited epithelial mesenchymal transition (EMT) in breast cancer cells. Taken together, our results revealed that KIF3B is up-regulated in breast cancer which is potentially involved in breast cancer progression and metastasis. Silencing KIF3B might suppress the Wnt/β-catenin signaling pathway and EMT in breast cancer cells.

Construction of SIRT1 gene shRNA lentivirus vector and its effect on the proliferation of breast cancer cells

Current experiment aimed to investigate the construction of the SIRT1 gene shRNA lentivirus vector and its effect on proliferation of breast cancer cells. Altogether 80 cases of breast cancer tissues and 80 cases of normal adjacent tissues were collected. qPCR was used for detecting SIRT1 expression. Western blot was used to detect the expression of EMT marker protein. The effect of lentivirus infected sh-SIRT1 on the cell biological function of SK-BR-3 and MDA-MB-231 cells was detected. MTT assay was used to detect cell activity, Transwell cell was used to detect cell invasion and migration, and cell apoptosis detected by flow cytometry. Compared with normal tissues adjacent to cancer, the expression of SIRT1 in cancer tissues increased significantly. Compared with human breast epithelial cells (MCF 10A), SIRT1 expression in breast cancer cells (MDA-MB-231, SK-BR-3) increased significantly. The above results showed that SIRT1 was significant greatly expressed in breast cancer. Compared with the sh-Control group, the cell activity, invasion and migration of the sh-SIRT1 group were enhanced, while cell apoptosis was weakened. In the sh-SIRT1 group infected by lentivirus, cell activity, cell invasion and migration decreased, while cell apoptosis increased. Compared with sh-Control, the expression of α-catenin, PTEN and E-cadherin in the sh-SIRT1 group in SK-BR-3 and MDA-MB-231 cells was down-regulated, while the expression of N- cadherin, β-catenin and Vimentin was up-regulated. Compared with sh-Control, the expression of α-catenin, PTEN and E-cadherin in the sh-SIRT1 group infected by lentivirus was up-regulated, while the expression of N- cadherin, β-catenin and Vimentin was down-regulated. To sum up, SIRT1 is highly expressed in breast cancer cells. The proliferation of breast cancer cells was inhibited after lentivirus infection with sh-SIRT1.

Abstract P6-06-18: Promyelocytic leukemia (PML) is induced by tumor-associated macrophages (TAMs) in breast cancer

Abstract Introduction: Tumor-associated macrophages (TAMs) are pivotal orchestrators of the tumor microenvironment. Clinical evidences show that density of TAMs increases with lymph node metastasis, which is associated with poor prognosis in breast cancer patients. The underlying mechanism has not been clearly elucidated. Methods: Transcriptome of MCF-7 cells cocultured with TAMs or monocyte-derived macrophages (MDMs) was analyzed and compared by RNA-seq analysis, which showed that the promyelocytic leukemia (PML) gene was significantly upregulated by TAMs (P&amp;lt;0.001). The upregulation of PML was further verified by qRT-PCR and immunohistochemistry (IHC) in tissue microarrays (TMAs) containing 294 cases of breast cancer tissue with duplicate cores and 28 cases of normal breast tissue with single core. The age, sex, pathological type, histologic grade and stage of the patients were well recorded. Results: RNA-seq analysis revealed that TAMs significantly upregulated PML expression, which was confirmed by qRT-PCR. Further, the protein level of PML was detected by IHC in TMAs. As expected, 57 cores of benign breast tissue, including normal breast tissue, adjacent normal breast tissue, adenosis and fibrous tissue in invasive ductal carcinoma (IDC) and lobular carcinoma in situ (LCIS), showed absent (80.70%) or low (19.30%) staining scores. In 508 (99.03%) cores of invasive breast cancer tissue, PML expression was positively correlated with lymph node metastasis (P= 0.003). However, no significant associations were observed between PML expression and tumor stage, primary tumor size, histologic tumor grade in IDC, or PML cellular localization. Conclusion:Recently, PML, upregulated in a subset of breast cancer, has been identified as a breast cancer oncogene. Here, we demonstrated PML expression was induced by TAMs and positively correlated with lymph node metastasis. Thus, we conclude PML mediates TAMs function in breast tumorigenesis and TAMs stimulates the expression of PML in breast cancer. Citation Format: Meijun Long, Mei Yang, Jun An, Renbin Liu. Promyelocytic leukemia (PML) is induced by tumor-associated macrophages (TAMs) in breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P6-06-18.

Good staining quality ensuring the reproducibility of Ki67 assessment

AimsAlthough Ki67 labelling index (LI) is a prognostic and predictive marker in breast cancer, its accuracy and reproducibility must be validated before its clinical application. We aimed to evaluate the...

Expression of Stanniocalcin 2 in Breast Cancer and Its Clinical Significance.

This study aims to explore the expression of stanniocalcin 2 (STC2) gene in breast cancer and its clinical significance. Female patients with breast cancer from Zhongnan Hospital of Wuhan University admitted during March 2014 to October 2014 were enrolled in this study. All the tissues used in this experiment included 50 cases of breast cancer tissues and corresponding 50 cases of paracancer normal breast tissues with complete patients' information. The real-time quantitative polymerase chain reaction (qPCR) was applied to detect the expression of STC2 gene in 50 cases of breast cancer and paracancer normal breast tissues. The results showed that the expression level of STC2 gene in 50 cases of breast cancer tissues was significantly higher than that in paracancer normal breast tissues (P&lt;0.001). The expression of STC2 gene was correlated with lymph node metastasis, distant metastasis, TNM stage and histological grade (P&lt;0.001). The expression level of STC2 gene was significantly higher in breast cancer tissues with higher expression of Ki-67 (P&lt;0.001). The expression level of STC2 gene was significantly higher in estrogen receptor (ER) positive breast cancer tissues than in ER negative ones (P&lt;0.001). However, different groups of age, pathological type, tumor size, PR expression and human epidermal growth factor receptor-2 (HER2) expression did not show significant differences in STC2 expression (P&gt;0.05). In conclusion, the abnormal overexpression of STC2 gene may play a role in the development and progression of breast cancer, and it can be used as an independent metastasis and prognostic factor of breast cancer. In addition, STC2 gene probably promotes the development and metastasis of breast cancer by interacting with estrogen and ER, and it may become a new direction for breast cancer endocrine therapy.

LncRNA DLEU1/microRNA-300/RAB22A axis regulates migration and invasion of breast cancer cells.

To explore the possible role of deleted in lymphocytic leukemia 1 (DLEU1) in regulating the metastasis of breast cancer and its underlying mechanism. The expression levels of DLEU1 in 60 cases of breast cancer tissues and adjacent normal tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Survival analysis and receiver operating characteristic (ROC) curves were introduced to analyze the correlation between DLEU1 expression and the clinical features of enrolled breast cancer patients. After altering expressions of DLEU1, RAB22A or microRNA-300 by plasmid transfection, the abilities of breast cancer cells to migrate and invade were evaluated by transwell assay. Western blot was conducted to detect protein level changes of epithelial-mesenchymal transition (EMT)-related genes regulated by DLEU1, RAB22A or microRNA-300. Dual-luciferase reporter gene assay was performed to detect the binding relation between microRNA-300 with DLEU1 and RAB22A. DLEU1 expression remains higher in breast cancer tissues than in normal adjacent tissues, and has a certain diagnostic value. The overall survival of breast cancer patients was negatively correlated with DLEU1 expression. Overexpression of DLEU1 or RAB22A, or microRNA-300 knockdown could enhance the migratory and invasive capacities, as well as increase EMT ofBT549 cells. On the contrary, knockdown of DLEU1 or RAB22A, or microRNA-300 overexpression in MCF-7 cells obtained the opposite trends of cellular behaviors. Dual-luciferase reporter gene assay confirmed that DLEU1 and RAB22A could bind to microRNA-300. Further verification showed that RAB22A expression was positively regulated by DLEU1, but negatively regulated by microRNA-300. Finally, we found that DLEU1 overexpression could reverse the inhibitory effects of microRNA-300 on proliferation, migration, and EMT of breast cancer cells. DLEU1 is highly expressed in breast cancer, which promotes migration, invasion, and EMT of breast cancer cells by targeting microRNA-300 to mediate RAB22A.

Combined high expression of CD47 and CD68 is a novel prognostic factor for breast cancer patients

BackgroundAvoiding the phagocytosis by tumor-associated macrophages (TAMs) is necessary for the growth and metastasis of solid tumors. CD47 binds to the receptor signal-regulatory protein-α (SIRP-α) on the macrophages to avoid normal phagocytosis. In this study, we evaluated the expression and prognostic significance of CD47 and CD68-labeled TAMs in breast cancer solid tumors.MethodsTwo hundred seventeen cases of breast cancer tissues and 40 cases of benign breast lesions were collected for immunohistochemical staining of CD47 and CD68.ResultsBoth of the CD47 and CD68 expression were significantly higher in breast cancer tissues (P < 0.001), and associated with multiple clinicopathological parameters in breast cancer (P < 0.05). However, CD47 or CD68 expression alone was not an independent predictor of poor DFS in multivariate survival analysis (P > 0.05). Interestingly, combined high expression of CD47 and CD68 (CD47highCD68high) not only had a significant association with advanced TNM stage, histological grade, LNM, ER status, PR status and recurrence (P < 0.05), but also displayed a poorer 5-DFS (P = 0.011). Strikingly, CD47highCD68high served as a novel independent prognostic factor for poor DFS compared to the expression of CD47 or CD68 alone (P = 0.045). Furthermore, our study also showed for the first time that the prognostic significance of CD47highCD68high not only in breast cancer in general, but also in hormone receptor-negative breast cancer in particular.ConclusionsCombined detection of CD47 and CD68 may provide guidance for the prognosis of breast cancer, especially hormone receptor-negative breast cancer.

Abstract 1112: CCDC102B promotes metastatic cascade in breast cancer by activating NF-κB via down-regulating RACK1

Abstract Background: Breast cancer is one of the leading causes of cancer death among women and the pathological status of axillary lymph nodes is an important predictor of prognosis. However, the mechanism involved in this early stage of metastasis remains largely unknown. Methods: We used microarray analysis to carry out differential genomics analyses between matched pairs of sentinel lymph node metastatic tissues and breast primary tumors. We found 182 up-regulated genes in metastatic tissues (fold change≥1.5). We used CRISPR/Cas9 gene editing system to knockout these genes, each with 6 unique sgRNAs, and screened in vivo by transplanting loss-of-function cell pool into immunocompromised mice. MAGeCK was used to analyze the screening. Immunohistochemistry was performed to examine protein expressions in 212 cases of breast cancer tissues using tissue microarray (TMA) technology. Gene and protein expressions were monitored by qPCR and western blot. The relationship between coiled-coil domain containing 102B (CCDC102B) and receptor for activated C kinase 1 (RACK1) was examined using Co-IP, mass spectrometry, nuclear protein extraction assay and dual-luciferase reporter assay. Functional studies were detected in vitro by cell proliferation, wound-healing and migration, and in vivo by lung metastasis in immunocompromised mice injected CCDC102B-overexpressed cells into tail vein. The primary biological functions and pathways related to CCDC102B were enriched by RNA sequencing. The regulation of NF-κB signaling by CCDC102B and RACK1 was detected using dual-luciferase reporter assay and western blot. Results: We identified CCDC102B through screening and found that it was significantly up-regulated in breast cancer patients with poor prognosis. The expression of CCDC102B were independent predictors in both DFS and OS (P=0.009; P=0.014). CCDC102B expression was found to promote the proliferation and migration of breast cancer cells MDA-MB-231 and BT549, and lung metastasis in mice. Moreover, we found that overexpression of CCDC102B decreased RACK1 expression whereas knockdown of CCDC102B increased RACK1 expression at both protein and mRNA levels, and the aggressive behavior of CCDC102B in breast cancer cells could be reversed by the expression of RACK1. According to gene set enrichment analysis (GSEA), CCDC102B was correlated with significant enrichment focused on NF-κB pathway. The mechanistic study revealed that gain of CCDC102B resulted in decreased expression of RACK1, which increased levels of epithelial mesenchymal transition (EMT) and phosphorylated Rel-A (p65), leading to the activation of NF-κB pathway and progression of breast cancer metastasis. Conclusion: Taken together, our findings suggest that CCDC102B is strongly associated with negative clinical outcomes and may serve as a potential metastasis promotor by regulating the activation of NF-κB pathway via RACK1. Citation Format: Jing Si, Rong Guo. CCDC102B promotes metastatic cascade in breast cancer by activating NF-κB via down-regulating RACK1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1112.

Crumbs protein homolog 3 (CRB3) expression is associated with oestrogen and progesterone receptor positivity in breast cancer.

The crumbs protein homolog 3 (CRB3) regulates the tight junction to help maintain epithelial polarity. Altered CRB3 expression was associated with carcinogenesis of epithelial cells. This study detected CRB3 expression in 192 cases of breast cancer tissues and in the Molecular Taxonomy of Breast Cancer International Consortium (Metabric) and The Cancer Genome Atlas (TCGA) datasets for association with triple negative breast cancer (TNBC) phenotypes. The in vitro experiments confirm the ex vivo data. The data showed that levels of both CRB3 mRNA and protein were associated with TNBC phenotypes, ie, 41.1% (39/95) of ER+ breast cancer was CRB3‐positive, whereas 26.9% (25/93) ER– tumour was CRB3‐positive (P = 0.046). Moreover, 47.6% (30/63) of PR+ breast cancer was CRB3‐positive vs 28.4% (33/116) PR‐ tumours positive for CRB3 (P = 0.013). In addition, 40.1% (27/66) of ER+/PR+ tumour was CRB3‐positive, but only 22.4% (19/85) of TNBC showed CRB3 expression (P = 0.048). Indeed, levels of CRB3 mRNA were higher in non‐TNBC than TNBC in both Metabric (P = 3.682e‐10) and TCGA datasets (P = 2.501e‐07). The in vitro data showed that CRB3 expression was higher in luminal (MCF7 and T47D) than in HER2 (MDA‐MB‐453 and SK‐BR‐3) and basal (MDA‐MB‐231 and BT‐549) breast cancer cell lines. More interestingly, ERα regulated expression of CRB3 protein in MCF7 and BT‐549 cells and ERα expression was associated with CRB3 expression in breast cancer tissues specimens. This study demonstrated that ERα could be a novel regulator for CRB3 expression in breast cancer.

LncRNA RP11-316M1.12 promotes invasion of breast cancer cells

Objective To explore the role and mechanism of long non-coding RNA (lncRNA) RP11-316M1.12 in breast cancer cells. Methods Bioinformatics analysis was performed to varify RP11-316M1.12 expression pattern in breast cancer tissues and normal tissues. Quantitative real time polymerase chain reaction (qRT-PCR) assay was used to check the expression level of RP11-316M1.12 in breast cancer cells and tissues. Further, the correlationship between RP11-316M1.12 expression and clinical paremeters of breast cancer was analysed according to RP11-316M1.12 level. RP11-316M1.12 was overexpressed in MCF-7 cells, and RP11-316M1.12 was knocked down in MDA-MB-231 cells. Transwell method was used to detect the invasive ability of these cells. Western blot was used to detect the expression of epithelial-mesenchymal transition (EMT) markers in these cells. Results Gene Expression Profiling Interactive Analysis (GEPIA) database suggested that RP11-316M1.12 was highly expressed in breast cancer tissues than that in normal tissues. The similar results were got in 65 cases of breast cancer tissues and 23 cases of normal tissues by qRT-PCR assay. Meanwhile, we found that RP11-316M1.12 was enhanced in breast cancer cells than that in normal epithelial cell and RP11-316M1.12 expression is related to TNM stage and distant metastasis in breast cancer. Transwell assay demonstrated that RP11-316M1.12 significantly enhanced breast cancer cells invasion. Mechanismly, over expression of RP11-316M1.12 can remakably downregulated E-cadherin, enhanced ZEB1 and Vimentin expression in these cells. Conclusions RP11-316M1.12 was enhanced in breast cancer, and RP11-316M1.12 could accelerate invasion of breast cancer cells. Key words: RNA, untranslated; Breast neoplasms; Cell migration assays; In vitro

TRIM59 overexpression correlates with poor prognosis and contributes to breast cancer progression through AKT signaling pathway.

TRIM59 has been recently implicated in the carcinogenesis of several cancers such as lung cancer, gastric cancer, and bladder cancer. However, its expression pattern and clinical significance has not been investigated in human breast cancer. In the present study, we examined TRIM59 protein expression in 95 cases of breast cancer tissues using immunohistochemistry. We found that TRIM59 was upregulated in 42 out of 95 cases and correlated with TNM stage (P = 0.0056), lymph node metastasis (P = 0.0088) and poor prognosis (P = 0.0092). Importantly, TRIM59 level was higher in triple-negative breast cancer (TNBC) (P = 0.0157). Expression of TRIM59 protein was also upregulated in breast cancer cell lines compared to normal MCF-10A cell line. TRIM59 plasmid and shRNA transfection was performed in MCF-7 and SK-BR-3 cells respectively. TRIM59 overexpression promoted cell proliferation, invasion, migration, cell cycle transition, and paclitaxel resistance, whereas TRIM59 depletion showed the opposite results. Further analysis showed that TRIM59 overexpression upregulated expression of cyclinA, cyclinE, Bcl-xl, Bcl-2, p-AKT, and downregulated expression of p21, p27, p53. AKT inhibitor treatment abolished the effect of TRIM59 on Bcl-2 expression. TRIM59 overexpression also upregulated the level of p53 ubiquitination. In conclusion, TRIM59 overexpression correlates with poor prognosis and promotes malignant behavior through regulation of AKT pathway in human breast cancer.

Relationship between expression of RBM38 and distant metastasis and prognosis of breast cancer

Objective: To investigate the relationship between the expression of RNA-binding protein 38 (RBM38) and distant metastasis and prognosis of breast cancer. Methods: A total of 161 cases of breast cancer tissues were selected, and the expression of RBM38 was detected by immunohistochemical SP method. The relationship between the expression of RBM38 and the distant metastasis and prognosis of breast cancer patients were analyzed. Cox regression model was applied to analyze the influence factors of distant metastasis of breast cancer. Results: Among the 161 breast cancer patients, high expression of RBM38 was observed in 85 cases and low expression was observed in 76 cases. Log rank test showed that the distant metastasis rate of RBM38 highly expressed group was significantly lower than that of RBM38 lowly expressed group (P=0.010), while the survival rate was significantly higher (P=0.009). Univariate Cox regression analysis showed that the pathological stage (P=0.001), axillary lymph node positive (P=0.001), and the expression of RBM38 (P=0.015) were dramatically related to the distant metastasis of breast cancer patients. Multivariate Cox regression analysis showed that positive axillary lymph node was an independent risk factor (RR=4.222, 95% CI: 1.235-14.432), and high expression of RBM38 was an independent protective factor (RR=0.328, 95% CI: 0.128-0.839) of distant metastasis of breast cancer patients. Conclusion: High expression of RBM38 is positively related with low distant metastasis rate and good prognosis of breast cancer patients. Hence, high expression of RBM38 is an independent protective factor of distant metastasis.

Prediction of tumor mutation burden in breast cancer based on the expression of ER, PR, HER-2, and Ki-67.

ObjectiveCancer immunoediting is the process of eliminating highly immunogenic tumor cells by somatic evolution and protecting the host from tumor development in the host immune system. Frequencies of somatic mutations or tumor mutation burden (TMB) were associated with immunogenicity of breast cancer. This study aimed to predict the level of TMB in patients with breast cancer by the expression of estrogen (ER), progesterone (PR), HER-2, and Ki-67, thereby anticipating the prognosis of patients and the possible response to immunotherapy.Patients and methodsIn 53 patients with breast cancer, the 453 multigenes panel based on NGS was used to determine the TMB value of breast cancer in the patient’s primary tumor tissues. The predicted TMB value was divided into 4 groups: A (0–3.33), B (3.33–5.56), C (5.56–8.89), and D (>8.89), according to the quartile method, with group A as reference level. Logistic regression was used to analyze the risk ratio of each molecule type, and the prediction model was established. Survival probabilities by covariates were assessed using Kaplan–Meier estimator survival analysis and Cox’s proportional hazards models.ResultsIn 53 patients, the TMB value measured by the NGS polygenic panel was between 0 and 14.4/Mb. TMB distribution in 53 cases of breast cancer tissue: 18 cases in A group, 22 cases in B group, 10 cases in C group, and 3 cases in D group. HER-2 expression positivity was significantly associated with TMB (HER-2 positive vs HER-2 negative, odds ratio [OR] =34.81, 95% confidence interval [CI]: 3.711–821.689, P=0.0065). Higher TMB was distributed in the patients who were Ki-67 expression positive (>14%) than those who were Ki-67 expression negative (≤14%) (OR =0.217, 95% CI: 0.054–0.806, P=0.0242). However, no significant differences of TMB were found between ER-positive group and ER-negative group (OR =3.133, 95% CI: 0.124–127.687, P=0.4954) and between PR-positive group and PR-negative group in terms of TMB (OR =1.702, 95% CI: 0.162–20.335, P=0.6492). The predicted model is TMB = −1.14×ER +0.53×PR +3.55×HER-2-1.53×Ki-67+ CONSTANT (INTERCEPT). Patients with low TMB had a better disease-free survival (DFS) than those with high TMB (83 vs 59 m, P=0.002). In a multivariate analysis, high TMB (>5.56) was an independent predictive factor for decreased DFS (adjusted hazard ratio [HR], 5.594; 95% CI: 1.694–18.473; P = 0.005).ConclusionThe preliminary results suggest that the level of TMB value in patients with breast cancer can be predicted based on the expression levels of ER, PR, HER-2, and Ki-67, which may indicate the prognostic and predictive value of immunotherapy in patients with breast cancer.

The expression of LTBP2 in breast cancer and its clinical significance

Objective: To explore the expression of latent transforming growth factor (TGF)-beta binding protein 2 (LTBP2) in breast cancer and its clinical significance. Methods: Immunohistochemistry was used to detect the expression of LTBP2 in 125 cases of breast cancer tissue and normal breast tissue. Relationship between the expression of LTBP2 and estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), Ki67, and its relationship with clinicopathological parameters of breast cancer were analyzed. Results: The positive expression of LTBP2 in breast cancer tissues (55.20%) was significantly higher than that in adjacent normal breast tissues (28.80%) (P<0.001). The expression of LTBP2 in breast cancer tissues was correlated with the clinical stage (χ(2)=4.516, P=0.034), over-expression of ER (χ(2)=7.112, P=0.008) and Ki67(χ(2)=4.024, P=0.045) and other adverse prognostic factors. Conclusion: Positive expression of LTBP2 in breast cancer plays an important role in the development and progression of breast cancer, and may be a marker of prognosis of patients.

Expression and clinical significance of OTUB1 in breast cancer

Objective To explore the expression level of OTUB1 and its clinical significance in breast cancer. Methods Immunohistochemistry was used to detect the expression level of OTUB1 in 78 cases of breast cancer tissues and 30 cases of normal breast tissue adjacent to carcinoma, and the relationships between OTUB1 and the clinical pathological features of breast cancer were analyzed. Results The positive expression rate of OTUB1 in breast cancer tissues [66.7% (52/78)] was significantly higher than that in adjacent normal breast tissues [30.0% (9/30)], with a statistically significant difference (χ2=11.851, P=0.001). OTUB1 expression level was related to the lymph node metastasis (χ2=5.029, P=0.025), postoperative TNM staging (χ2=4.478, P=0.034), expression of human epidermal growth factor receptor-2 (HER-2) (χ2=8.775, P=0.003), expression of P53 (χ2=4.708, P=0.030), expression of estrogen receptor (ER) (χ2=10.364, P=0.001) and molecular subtypes (χ2=10.934, P=0.012). However, OTUB1 expression level in breast cancer was not related to the age (χ2=2.194, P=0.139), menopausal status (χ2=1.843, P=0.175), tumor size (χ2=0.643, P=0.423), histological grade (χ2=3.580, P=0.167), expression of progestin receptor (PR) (χ2=3.371, P=0.066) and expression of Ki-67 (χ2=1.345, P=0.246). Conclusion OTUB1 expression level increases in breast cancer, which is associated with the lymph node metastasis, TNM staging, expressions of HER-2, P53, ER and molecular subtypes of breast cancer. It suggests that the expression of OTUB1 is related to the progression and metastasis of breast cancer. Key words: Breast neoplasms; Immunohistochemistry; OTUB1

Expression and clinical significance of SATB1 and TLR4 in breast cancer.

This study investigated the expression of special AT-rich sequence-binding protein 1 (SATB1) and toll-like receptor 4 (TLR4) protein in breast cancer and its clinical significance. We collected breast cancer tissues from 120 patients and adjacent non-cancerous tissue from 53 patients. SATB1 was expressed in 89 cases of breast cancer (74.17%) and in 7 cases of adjacent non-cancerous tissue (13.21%). TLR4 was expressed in 70 cases of breast cancer tissues (58.33%) and in 48 cases of adjacent non-cancerous tissue (90.57%). The differences of SATB and TLR4 in breast cancer and adjacent non-cancerous tissue were statistically significant. We found a negative correlation between the expression of SATB1 and TLR4 (r=−0.624, P<0.05). The expression of SATB1 and TLR4 were not significantly correlated with age, menopause, and PR and HER-2 protein expression, but were significantly correlated with tumor size, local lymphatic metastasis, histopathological grade, tumor stage, and ER protein expression (P<0.05). Overall, SATB1 and TLR4 proteins are involved in the development of breast cancer, a finding of great significance to identify therapeutic targets and prognosis markers for breast cancer.

LncRNA MEG3 inhibits cell epithelial-mesenchymal transition by sponging miR-421 targeting E-cadherin in breast cancer

BackgroundMEG3, a lncRNA, has been verified in several tumors to function as tumor suppressors including breast cancer development and progression, however, the expression pattern and underlying mechanisms of MEG3 involved in breast cancer progression is still need to be further explored. MethodsThe expression of MEG3 was confirmed in 90 cases of breast cancer tissues compared to adjacent normal tissues by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The association between clinicopathological factors and MEG3 expression was evaluated by chi-square test. Kaplan–Meier curve and log rank test was performed to assess disease-free survival (DFS) and overall survival (OS) time in patients. CCK8 and transwell invasion assays were used to assess cell proliferation and invasion capacity. Luciferase report assay and RNA pull down assay were used to detect the association between miR-421 and MEG3 in breast cancer. ResultsIn the study, we demonstrated that the expression of MEG3 was significantly down-regulated in breast cancer tissues compared to adjacent normal tissues. Reducing MEG3 expression was significantly associated with TNM stage and lymph nodes metastasis in patients. Survival analysis showed that lower MEG3 predicted a poor DFS and OS for patients. In vitro, we showed that up-regulated MEG3 inhibited cell proliferation and cell invasion capacities. We further revealed that endogenous miR-421 expression was negatively regulated by MEG3 in breast cancer cells and MEG3 regulated E-cadherin expression by sponging to miR-421 in breast cancer cells. ConclusionsOur results showed that MEG3/miR-421/E-cadherin regulatory axis may be a novel therapeutic target for breast cancer.