A Novel miRNA Y-56 Targeting IGF-1R Mediates the Proliferation of Porcine Skeletal Muscle Satellite Cells Through AKT and ERK Pathways.

As a key regulator of gene transcription and post-transcriptional modification, miRNAs play a wide range of roles in skeletal muscle development. Skeletal muscle satellite cells contribute to postnatal growing muscle fibers. Thus, the goal of this study was to explore the effects of novel miRNA Y-56 on porcine skeletal muscle satellite cells (PSCs). We found that Y-56 was highly expressed in porcine muscle tissues, and its expression was higher in Bama Xiang pigs than in Landrace pigs. The EdU assay, cell counting kit-8, and flow cytometry results showed that Y-56 overexpression suppressed cell proliferation and cell cycle, whereas Y-56 inhibition resulted in the opposite consequences. The results of qRT-PCR and Western blot showed that Y-56 remarkably inhibited the expression levels of cyclin-dependent kinase 4 (CDK4), proliferating cell nuclear antigen (PCNA), and cyclin D1. We identified that IGF-1R was a direct target of Y-56 by dual-luciferase reporter assay. Moreover, IGF-1R overexpression promoted the proliferation and cell cycle process of PSCs and upregulated the expression of CDK4, PCNA, and cyclin D1. Conversely, IGF-1R knockdown had the opposite effect. Furthermore, IGF-1R overexpression partially reversed the inhibition of the cell proliferation and cell cycle process of PSCs and the downregulation of the expression of CDK4, PCNA, and Cyclin D1 caused by Y-56 overexpression. Finally, Y-56 inhibited the protein expression levels of p-AKT and p-ERK. Collectively, our findings suggested that Y-56 represses the proliferation and cell cycle process of PSCs by targeting IGF-1R-mediated AKT and ERK pathways.

Balancing Between Polarization and Conduction Loss toward Strong Electromagnetic Wave Absorption of Hard Carbon Particles with Morphology Heterogeneity.

The heterostructure and hierarchical morphology of carbonaceous absorbents play an important role in the construction of high-performance electromagnetic wave absorbing materials. Herein, novel micron-scale hard carbon particles with morphology heterogeneity were developed as lightweight superior electromagnetic wave absorbents via a facile and ecofriendly process. The as-prepared hard carbon particle composed of pseudographite and a highly disordered region shows a unique heterostructure. Concurrently, constructing a multilevel geometric shape and size can cause a decrease of the percolation threshold and an excellent balance between polarization and conduction loss, which enhances the electromagnetic wave absorption significantly. The composites (thickness d = 2.36 mm) filled with morphology-heterogeneity hard carbon particles (15 wt %) achieve an excellent electromagnetic wave absorption with a minimum reflection loss of -78.0 dB at 10.2 GHz and effective absorption bandwidth (<-10 dB) of 3.1 GHz (8.8-11.9 GHz). Compared to the traditional carbonaceous absorbents with complex microstructures and/or multiple chemical components, this work presents a feasible idea for the development of an efficient carbonaceous absorbent to realize practical applications.

DRAIC promotes growth of breast cancer by sponging miR-432-5p to upregulate SLBP.

Mounting evidence suggests that lncRNAs can exert functions in cancer progression in multiple manners. In recent years, competing endogenous RNA (ceRNA) has been widely reported in human cancers as a lncRNA-dominant molecular pathway. The current study aimed at proving the role of lncRNA downregulated RNA in cancer (DRAIC) in breast cancer (BRCA) progression. To be specific, qRT-PCR assay was conducted to measure the expression of DRAIC and other downstream target genes. It was uncovered that DRAIC was expressed at a high level in BRCA cells. Functional analyses, including CCK-8, colony formation, and EdU assays demonstrated that DRAIC depletion suppressed BRCA cell proliferation. In addition, cell apoptosis was promoted due to DRAIC knockdown. The inhibitory effect of DRAIC reduction on BRCA cell migration and invasion was proven by transwell assays. Mechanistically, DRAIC was confirmed to predominantly distribute in the cytoplasm and could interact with miR-432-5p. In addition, stem-loop binding protein (SLBP) was verified to be a downstream target of miR-432-5p and was positively regulated by DRAIC. Taken together, DRAIC sponged miR-432-5p to enhance SLBP expression, by which malignant behaviors of BRCA cells were promoted. Our findings may help to provide a promising therapeutic target for BRCA patients.

LINC01234 aggravates cell growth and migration of triple-negative breast cancer by activating the Wnt pathway.

Triple-negative breast cancer (TNBC) is a common cancer with increasing incidence and mortality in female. Increasing studies have revealed that long noncoding RNAs (lncRNAs) are novel molecules regulating tumors. Long intergenic non-protein coding RNA 1234 (LINC01234) has been demonstrated to function as an oncogene in several tumors. However, the role of LINC01234 in TNBC remains unelucidated. Herein, RT-qPCR showed that LINC01234 expression was upregulated in both TNBC tissues and cell lines. Functionally, knockdown of LINC01234 suppressed proliferation, migration, invasion, epithelial-mesenchymal transition (EMT) process, and promoted apoptosis in TNBC cells. Xenograft mouse models revealed that LINC01234 downregulation inhibited TNBC tumor growth in vivo. Furthermore, LINC01234 was transcriptionally elevated by Sp1 transcription factor (SP1) in TNBC cells. Mechanistically, LINC01234 interacted with miR-525-5p and miR-525-5p targeted MEIS2. Rescue assays manifested that MEIS2 overexpression rescued the cellular processes inhibited by silenced LINC01234. Moreover, we validated that LINC01234 regulated the activation of the Wnt pathway through modulating MEIS2 in TNBC cells. In conclusion, LINC01234 aggravated TNBC cell growth, migration, invasion and EMT by modulating the miR-525-5p/MEIS2 axis and activating the Wnt/ß-catenin signaling pathway.

Circ_0000511 accelerates the proliferation, migration and invasion, and restrains the apoptosis of breast cancer cells through the miR­326/TAZ axis.

Circular RNA 0000511 (circ_0000511) has been observed to be dysregulated in breast cancer (BC). However, the functions of circ_0000511 in breast cancer remain unknown. The expression levels of circ_0000511, ribonuclease P RNA component H1, microRNA­326 (miR­326) and transcriptional co­activator with PDZ­binding motif (TAZ) were examined by reverse transcription­quantitative PCR. Colony formation and MTT assays were conducted to analyze the cellular proliferative ability. The apoptotic rate was assessed by flow cytometry. Western blot analysis was used to evaluate the expression levels of B cell leukemia/lymphoma 2 (Bcl­2), Bcl­2 associated X apoptosis regulator, cleaved caspase­3 and TAZ. Transwell assays were performed to evaluate the migration and invasion of BC cells. The target interaction between miR­326 and circ_0000511 or TAZ was confirmed by dual­luciferase reporter assay. Xenograft assay was used to identify the function of circ_0000511 in vivo. Circ_0000511 abundance was abnormally elevated in BC tissue samples and cell lines compared with in matched normal cases. Circ_0000511 interference suppressed the proliferation, migration and invasion, and induced apoptosis of BC cells. miR­326 was a direct target of circ_0000511, and circ_0000511 silencing­mediated effects in BC cells were largely reversed by the knockdown of miR­326. miR­326 directly bound to TAZ mRNA, and TAZ accumulation largely attenuated miR­326 overexpression­induced effects in BC cells. Circ_0000511 upregulated the expression levels of TAZ partly via targeting miR­326 in BC cells. Circ_0000511 silencing restrained tumor growth in vivo. Circ_0000511 accelerated the proliferation, migration and invasion, while inhibiting the apoptosis of BC cells through upregulating TAZ expression via sponging miR­326. The circ_0000511/miR­326/TAZ axis may be a novel therapeutic target for BC treatment.

STAT3-induced HLA-F-AS1 promotes cell proliferation and stemness characteristics in triple negative breast cancer cells by upregulating TRABD.

Breast cancer (BC) is one of the most common malignances and is a leading cause of cancer-related deaths in women globally. Triple negative breast cancer (TNBC) is a common subtype of BC. Emerging evidence has indicated the crucial roles of long noncoding RNAs (lncRNAs) in the tumorigenesis of TNBC. Our aim was to explore the role and regulatory mechanism of lncRNA HLA-F antisense RNA 1 (HLA-F-AS1) in TNBC cells. Cell counting kit-8 (CCK-8) assay, colony formation assay, flow cytometry analysis and western blot analysis were used to measure HLA-F-AS1-mediated cellular behaviors in TNBC. Xenograft tumor assay was applied to assess biological function of HLA-F-AS1 in vivo. Luciferase reporter assay and RNA pull down assay were used to verify the binding ability between molecules. Our findings demonstrated that HLA-F-AS1 expression was significantly upregulated in TNBC tissues and cells, and high level of HLA-F-AS1 indicated the poor prognosis of patients with TNBC. HLA-F-AS1 promoted TNBC progression by facilitating cell proliferation and stemness maintenance and inhibiting cell cycle arrest at G0/G1 stage and apoptosis in vitro as well as inducing tumor growth in vivo. HLA-F-AS1. In addition, signal transducer and activator of transcription 3 (STAT3) transcriptionally induced HLA-F-AS1 upregulation in TNBC cells via interacting with HLA-F-AS1 promoter. Moreover, HLA-F-AS1 acted as the molecular sponge of microRNA 541-3p (miR-541-3p) to elevate TRABD (TraB domain containing) expression in TNBC cells. Rescue experiments confirmed that the decrease of cell proliferation and stemness characteristics under silenced HLA-F-AS1 was rescued by TRABD overexpression in TNBC cells. In conclusion, STAT3-induced HLA-F-AS1 facilitates cell proliferation and stemness characteristics in TNBC by miR-541-3p-dependent upregulation of TRABD, which might provide a potential novel direction for the treatment of TNBC.

Regulatory effect of the MAFG­AS1/miR­150­5p/MYB axis on the proliferation and migration of breast cancer cells.

Long noncoding RNA (lncRNA) MAF BZIP transcription factor G antisense RNA 1 (MAFG­AS1) has been demonstrated to serve an important role in the progression of various types of cancer, whereas its role in breast cancer has not been fully elucidated. The present study aimed to explore the potential role and underlying mechanism of MAFG­AS1 in breast cancer. To achieve this, the expression of MAFG­AS1, microRNA (miR)­150­5p and MYB was detected by reverse transcription­quantitative PCR. The binding between miR­150­5p and MAFG­AS1 or MYB was verified using a luciferase reporter assay. Cell proliferation was analyzed by MTS, apoptosis and cell cycle were detected by Annexin V/propidium iodide, and cell migration was analyzed by wound healing assay. The results demonstrated that the expression levels of MAFG­AS1 were significantly upregulated in breast cancer tissues and cells compared with those in normal breast tissues and cells. High MAFG­AS1 expression promoted the proliferation, migration and epithelial­mesenchymal transition of breast cancer cells. By contrast, miR­150­5p expression was reduced in breast cancer tissues compared with that in healthy breast tissues, and low expression of miR­150­5p was associated with poor overall survival in patients with breast cancer. Bioinformatics and luciferase assay revealed that MAFG­AS1 served as a sponge of miR­150­5p, and that miR­150­5p bound to MYB. The functional rescue assay results demonstrated that MAFG­AS1 knockdown suppressed the proliferation and migration of breast cancer cells by regulating miR­150­5p, which in turn targeted MYB. In conclusion, the results of the present study demonstrated that MAFG­AS1 functioned as a novel oncogenic lncRNA in the development of human breast cancer via regulating the miR­150­5p/MYB axis, which suggested that MAFG­AS1 may be a novel biomarker for the diagnosis and prognosis of human breast cancer.

Circ-PRMT5 promotes breast cancer by the miR-509-3p/TCF7L2 axis activating the PI3K/AKT pathway.

BACKGROUND: Breast cancer is the most prevalent malignancy occurring in females. In recent years, emerging evidence has suggested that circular RNAs are involved in the development of multiple cancers. Circ-PRMT5 has recently attracted attention as a tumor-promoting circular RNA. In the present study, we focused on exploring the biological effects of circ-PRMT5 in breast cancer. METHODS: A quantitative real-time polymerase chain reaction was used to determine the expression of circ-PRMT5 in breast cancer. In vitro experiments, including cell-counting kit-8, 5-ethynyl-2'-deoxyuridine, flow cytometry and tube formation assays, were performed to test the effects of circ-PRMT5 on the cellular progression of breast cancer. Bioinformatic analysis, luciferase reporter, radioimmunoprecipitation and RNA-pull down assays were performed to predict the potential microRNAs interacting with circ-PRMT5 and mRNAs that can be targeted by miR-509-3p. RESULTS: Circ-PRMT5 is up-regulated in breast cancer tissues and cells. Importantly, an elevation of circ-PRMT5 indicates a poor prognosis in patients with breast cancer. Functionally, knockdown of circ-PRMT5 suppresses cell proliferation and angiogenesis and increases cell apoptosis in breast cancer. Mechanistically, we identified that circ-PRMT5 up-regulates TCF7L2 expression by acting as a miR-509-3p sponge. The negative expression correlation between miR-509-3p and circ-PRMT5 or TCF7L2 in clinical tissues was further demonstrated. Rescue assays showed that TCF7L2 overexpression reverses the antitumoral effects of circ-PRMT5 knockdown on breast cancer cell processes. Additionally, we demonstrated that circ-PRMT5 activates the phosphoinositide 3-kinase (PI3K)/AKT pathway by up-regulation of TCF7L2. CONCLUSIONS: Overall, our data indicate that the circ-PRMT5/miR-509-3p/TCF7L2 axis can aggravate the malignant character of breast cancer cells by the regulation of the PI3K/AKT pathway.

Capsaicin suppresses breast cancer cell viability by regulating the CDK8/PI3K/Akt/Wnt/ß­catenin signaling pathway.

Breast cancer displays high morbidity and mortality. Despite exerting certain effects, traditional treatments cannot eliminate every cancer cell and may kill normal cells due to inaccurate targeting. However, as a traditional Chinese medicine, capsaicin, an active compound extracted from chili peppers, has displayed potent anticarcinogenic activities in vitro and in vivo, but the underlying mechanism is not completely understood. The pharmacological effects of capsaicin on tumors was evaluated in MDA MB 231 breast cancer cells. The MTT, cell scratch assay, cell cycle analysis, cell transfection, reverse transcription­quantitative PCR and western blotting were performed to investigate the potential antitumor mechanisms of capsaicin. In the present study, the potential anticancer mechanism underlying capsaicin in MDA­MB­231 cells in vitro was investigated. Capsaicin significantly inhibited MDA­MB­231 breast cancer cell viability and migration compared with the control group. The flow cytometry results indicated that capsaicin induced G2/M cell cycle arrest in MDA­MB­231 cells. In addition, capsaicin significantly reduced the expression of cyclin­dependent kinase 8 (CDK8) in breast cancer cells compared with the control group. Moreover, LV­CDK8 small interfering RNA­transduced MDA­MB­231 cells displayed lower CDK8 mRNA and protein expression levels compared with LV­negative control­shRNA­transduced cells. Furthermore, capsaicin significantly reduced the expression levels of phosphorylated (p)­PI3K, p­Akt, Wnt and ß­catenin in vitro compared with the control group. Collectively, the results of the present study suggested that capsaicin inhibited breast cancer cell viability, induced G2/M cell cycle arrest, reduced CDK8 expression levels, decreased the phosphorylation of PI3K and Akt and downregulated Wnt and ß­catenin expression levels in MDA­MB­231 cells.

RETRACTED: LncRNA GAS6-AS1 facilitates the progression of breast cancer by targeting the miR-324-3p/SETD1A axis to activate the PI3K/AKT pathway.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This article has been retracted at the request of the current Editor in Chief, with the agreement of all authors. The article is being retracted due to inappropriate manipulation of the original data for presentation in Figures 2C, 5C and 6C. Upon further investigation and discussion with the authors, insufficient evidence was provided to support a reasonable explanation for these mistakes. In addition, the authors could not provide convincing original data supporting other figures of the manuscript, e.g. Figures 2G, 4 and 6A. Based on these findings, the Editor in Chief and the authors have decided to retract the study. The authors apologize for any inconvenience caused.

TCF3-activated FAM201A enhances cell proliferation and invasion via miR-186-5p/TNKS1BP1 axis in triple-negative breast cancer.

Increasing evidence shows that long non-coding RNAs (lncRNAs) are closely associated with the development of cancers, including triple-negative breast cancer (TNBC). LncRNA FAM201A has been identified as a key regulator in some cancers. However, its role has not been explored in TNBC. In this work, we investigated the biological role and regulatory mechanism of FAM201A in TNBC. The expression pattern of FAM201A was determined by RT-qPCR analysis. The biological effect of FAM201A on cellular process of TNBC was tested using colony formation, EdU, caspase-3 activity detection, flow cytometry, wound healing, and Transwell assays. ChIP and luciferase reporter assays were performed to verify the interaction between transcription factor 3 (TCF3) and FAM201A. The interaction among FAM201A, microRNA-186-5p (miR-186-5p), and tankyrase 1 binding protein 1 (TNKS1BP1) was evaluated by luciferase reporter and RIP assays. The results showed that FAM201A expression was significantly upregulated in TNBC tissues and cells. Functionally, FAM201A knockdown inhibited TNBC cell proliferation, migration and invasion, and accelerated cell apoptosis. In mechanism, it was confirmed that FAM201A was transcriptionally activated by TCF3 and served as a sponge for miR-186-5p to upregulate TNKS1BP1 expression in TNBC cells. Collectively, our study revealed that TCF3-activated FAM201A promoted aggressive phenotypes of TNBC cells by upregulating TNKS1BP1 expression.

POU4F1 confers trastuzumab resistance in HER2-positive breast cancer through regulating ERK1/2 signaling pathway.

Over-expression of the human epidermal growth factor receptor-2 (HER2) is related to aggressive tumors and poor prognosis in breast cancer. Trastuzumab (TRA) resistance leads to tumor recurrence and metastasis, resulting in poor prognosis in HER2-positive breast cancer. POU Class 4 Homeobox 1 (POU4F1) is a member of the POU domain family transcription factors, and has a key role in regulating cancers. However, its effects on TRA-resistant HER2-positive breast cancer are still vague. In the present study, we found that POU4F1 expression was dramatically increased in clinical breast cancer specimens with TRA resistance. Higher POU4F1 was also detected in HER2-positive breast cancer cells with TRA resistance than that of the parental ones. Poor prognosis was detected in breast cancer patients with high POU4F1 expression. Under TRA treatment, POU4F1 knockdown significantly reduced the proliferative capacity of HER2-positive breast cancer cells with TRA resistance. POU4F1 silence also sensitized resistant HER-positive breast cancer cells to TRA treatment in vivo using a xenograft mouse model, along with the markedly reduced tumor growth rate and tumor weight. Moreover, we found that POU4F1 deletion greatly decreased the activation of mitogen-activated or extracellular signal-regulated protein kinase kinases 1 and 2 (MEK1/2) and extracellular-regulated kinase 1/2 (ERK1/2) signaling pathways in breast cancer cells with TRA resistance. Migration and invasion were also effectively hindered by POU4F1 knockdown in TRA-resistant HER2-positive breast cancer cells. Notably, we found that POU4F1 deletion-improved chemosensitivity of HER2-positive breast cancer cells with drug-resistance to TRA treatment was closely associated with the blockage of ERK1/2 signaling. Collectively, our findings reported a critical role of POU4F1 in regulating TRA resistance, and demonstrated the underlying molecular mechanisms in HER2-positive breast cancer. Thus, POU4F1 may be a promising prognostic and therapeutic target to develop effective treatment for overcoming TRA resistance.

Long non-coding RNA LRRC75A-AS1 facilitates triple negative breast cancer cell proliferation and invasion via functioning as a ceRNA to modulate BAALC.

As a common female malignancy, triple-negative breast cancer (TNBC) is the most serious subtype in breast cancer (BC). BAALC binder of MAP3K1 and KLF4 (BAALC) is a common oncogene in acute myelocytic leukemia (AML). We sought to explore the role of BAALC in TNBC. In this study, BAALC was significantly upregulated in TNBC tissues and cells. Then, the results of functional assays disclosed that BAALC facilitated cell proliferation, invasion, and epithelial-mesenchymal transition (EMT) processes, but repressed cell apoptosis in TNBC. Next, miR-380-3p was identified as the upstream of BAALC in TNBC cells. Moreover, LRRC75A-AS1 (also named small nucleolar RNA host gene 29: SNHG29) was verified to act as the sponge of miR-380-3p to elevate BAALC expression in TNBC. Besides, LRRC75A-AS1 could negatively regulate miR-380-3p but positively regulate BAALC expression. Finally, rescue assays elucidated that LRRC75A-AS1 facilitated cell proliferation, invasion, and EMT processes in TNBC by targeting miR-380-3p/BAALC pathway. Taken together, our study revealed a novel ceRNA network of LRRC75A-AS1/miR-380-3p/BAALC in accelerating TNBC development, indicating new promising targets for TNBC treatment.

Opening label, dynamic prospective cohort study on the small focus less than 1.0 cm shown by type B ultrasound in breast.

BACKGROUND: A consensus has not been achieved regarding the treatment of small nonpalpable breast lesions, and the purpose of this study was to prospectively investigate nonpalpable lesions less than 1.0 cm in diameter to explore the risk factors for such lesions and determine appropriate treatment of such kind of lesions. METHODS: A total of 1039 patients with small lesions less than 1.0 cm in diameter who underwent mammography and ultrasound from 2009 to 2010 in our institution were prospectively enrolled. Among them, 80 patients underwent biopsy, whose lesions grew by more than 30% of its original size, with an unclear boundary or irregular shape. All patients were followed-up for an average of 24 months, and lesions identified as high-risk types, such as cancer or atypical hyperplasia, of tumors on pathological examination were labeled "meaningful lesions." Then relevant factors affecting the detection of meaningful lesions were analyzed. RESULTS: In total, 40 meaningful lesions including 2 breast cancers were detected, accounting for 3.8% and 0.2% of all patients, respectively. Univariate analysis identified smoking (P = .030), irregular shape (P = .018), unclear boundary (P = .024), and vascularization (P = .023) as risk factors for the detection of meaningful lesions (P < .05). On multivariate analysis, smoking and irregular shape were further identified as independent risk factors for the detection of meaningful lesions. CONCLUSION: The overall incidence of cancer among nonpalpable lesions with a diameter less than 1.0 cm is low. Biopsies are strongly recommended for patients who are smokers or who have small lesions with an irregular shape, whereas regular follow-up observation is likely safe for other patients with small, non-palpable breast lesions.

A prognostic nomogram for overall survival in male breast cancer with histology of infiltrating duct carcinoma after surgery.

OBJECTIVE: The study was designed to construct and validate a nomogram for predicting overall survival (OS) of male breast cancer (MBC) patients with infiltrating duct carcinoma (IDC). METHODS: The cohort was selected from the Surveillance, Epidemiology, and End Results (SEER) database between January 1, 2004 and December 31, 2013. Univariate and multivariate Cox proportional hazard (PH) regression models were performed. A nomogram was developed based on the significant prognostic indicators of OS. The discriminatory and predictive capacities of nomogram were assessed by Harrell's concordance index (C-index), calibration plots, area under the curve (AUC) and the decision curve analysis (DCA). RESULTS: The median and maximal survival time of 1862 eligible patients were 49 and 131 months, respectively. Multivariate analysis showed that age (P < 0.0001), marital status (P = 0.002), T stage (P < 0.0001), N stage (P = 0.021), M stage (P < 0.0001), progesterone receptor (PR) (P = 0.046), human epidermal growth factor receptor-2 (HER2) (P = 0.009), and chemotherapy (P = 0.003) were independent prognostic indicators of IDC of MBC. The eight variables were then combined to construct a 3-and 5-year nomogram. The C-indexes of the nomogram were0.740 (95% confidence interval [CI] [0.709-0.771]) and 0.718 (95% CI [0.672-0.764]) for the internal validation and external validation, respectively. A better discriminatory capacity was observed in the nomogram compared with the SEER summary stage (P < 0.001) and AJCC TNM staging systems (6th edition; P < 0.001) with respect to OS prediction. Good consistency was detected between the nomogram prediction and actual findings, as indicated by calibration curves. The AUC for 3-and 5-year OS was 0.739 (95% CI [0.693-0.786]) and 0.764 (95% CI [0.725-0.803]) in the training cohort and 0.737 (95% CI [0.671-0.803]) and 0.735 (95% CI [0.678-0.793]) in the validation cohort, respectively. The DCA demonstrated that the survival nomogram was clinically useful. CONCLUSIONS: The nomogram was able to more accurately predict 3-and 5-year OS of MBC patients with IDC histology than were existing models.

Prevalence of BRCA1 and BRCA2 gene mutations in Chinese patients with high-risk breast cancer.

BACKGROUND: Breast cancer is the most common cancer among women worldwide. Here, we report the prevalence of BRCA1/2 mutations in patients with high-risk breast cancer from Inner Mongolia and Jilin, China, which was a part of a nationwide project on the detection of BRCA1/2 mutations in Chinese patients with hereditary breast cancer. METHODS: According to the criteria, index patients from a total of 245 independent families were initially recruited. All 49 exons of BRCA1 and BRCA2 and adjacent noncoding regions were screened for mutations based on next-generation sequencing from collected saliva. RESULTS: We detected 17 BRCA1/2 variants in 18 of 216 (8.3%) index patients with high-risk breast cancer. Among these, seven mutations were novel, including four BRCA1 mutations (c.123_124delCAinsAT, c.5093_5096delCTAA, c.5396-2A>G, and c.2054delinsGAAGAGTAACAAGTAAGAAGAGTAACAAGAAG), and three BRCA2 mutations (c.304A>T, c.7552_7553insT, and c.9548_9549insA). The BRCA1/2 variants were identified in 14% (8/57) of the patients with triple-negative breast cancer and in 6.3% (10/159) of the patients with non-triple-negative breast cancer. There was no significant difference between the two groups (p = 0.07). A higher frequency for BRCA1 mutations was observed in patients with triple-negative breast cancer than in those with non-triple-negative breast cancer (12.3% vs. 2.5%, p = 0.004). The frequencies of the BRCA2 mutations were not significantly different between patients with triple-negative breast cancer and those with non-triple-negative breast cancer (1.8% vs. 3.8%, p = 0.46). CONCLUSION: We found that patients with triple-negative breast cancer had a higher frequency of BRCA1 mutations than those with non-triple-negative breast cancer. In this study, no significant associations between the BRCA1/2 mutation status and age, family history of breast cancer, ovarian cancer, pancreatic cancer and prostate cancer, number of primary lesions, tumor size, or lymph node metastasis were observed.

Identification of claudin-2, -6, -11 and -14 as prognostic markers in human breast carcinoma.

The development of cancer occurs with various genomic and epigenetic modifications that act as indicators for early diagnosis and treatment. Recent data have shown that the abnormal expression of the claudin (CLDN) tight junction (TJ) proteins is involved in the tumorigenesis of numerous human cancers. Real-time quantitative PCR and western blotting were used to explore the differences in the expression of the CLDN TJ proteins in breast carcinoma tissues and non-neoplastic tissues. The results showed that CLDN5, CLDN9, CLDN12 and CLDN13 were not expressed in breast carcinoma tissues or non-neoplastic tissues. CLDN1, CLDN3, CLDN8 and CLDN10 were expressed in breast carcinoma and non-neoplastic tissues, but there was no significant difference between the expression of these CLDN proteins among them. The expression of CLDN2, -6, -11 and -14 varied between the breast carcinoma and non-neoplastic tissues. Moreover, 86 samples of breast carcinoma and non-neoplastic tissues were examined for the expression of CLDN2, -6, -11 and -14 by streptavidin-peroxidase immunohistochemical staining. The data revealed that the CLDN2, CLDN6, and CLDN14 were expressed in the cell membrane and the expression levels of these proteins were downregulated in breast carcinoma. The CLDN11 was expressed in cell cytoplasm and the expression level of CLDN11 was upregulated compared with those in non-neoplastic tissues. Consistent with these findings, the expression of CLDN2, CLDN6 and CLDN14 were downregulated, while the expression of CLDN11 was upregulated in breast carcinoma compared with those in non-neoplastic tissues. Furthermore, the associations between these CLDNs and clinicopathologic indicators were analyzed, and these CLDN expressions were revealed to be associated with distant metastasis and to predict a poor prognosis. In conclusion, our data showed that the expression levels of CLDN2, -6, -11 and -14 differed between breast carcinoma tissues and histologically non-neoplastic tissues, and the expression levels of these CLDNs may be useful as molecular markers for the diagnosis of breast carcinoma as well as for the determination of metastasis and prognosis.

Linkage disequilibrium and functional analysis of PRE1 insertion together with SNPs in the promoter region of IGFBP7 gene in different pig breeds.

Polymorphisms in regions upstream of transcription initiation site may modify the transcriptional activity of target genes by changing promoter activity. This study aims to determine whether or not polymorphisms at porcine IGFBP7 promoter region affect gene expression. In this study, eight SNPs and one PRE1 insertion in this region were first confirmed. The PRE1 insertion was widespread in 20 Chinese indigenous breeds, but was not observed in three commercial breeds. A perfect linkage disequilibrium, consisting of six of those SNPs and a PRE1, was observed with two haplotypes (h1 and h2) in five pig breeds. The h1 haplotype had an overwhelming superiority distribution in Large White, Landrace, and Bama mini-pig; in turn, the h2 only existed in the PRE1 presence breeds. As the haplotypes and PRE1 were located at gene promoter regions, we further investigated the transfection of plasmids with three different fragments of IGFBP-7 promoter region (H1, H2, RF). The CMV promoter of the pEGFP-N1 was substituted by these three different fragments, respectively. Different transcriptional and translational activities of EGFP in PK-15 cells were observed in these three recombinant plasmids by quantitative real-time PCR and flow cytometric analysis. The results indicated that H1 had the higher transcriptional and translational activities of EGFP as compared to the H2 (P < 0.05, P < 0.05). As compared to the RF group, EGFP mRNA expression level was significantly higher in H1 groups (P < 0.05). The IGFBP-7 promoter polymorphisms detected in this study may be important functional variants and potential genetic markers for pig population genetic study.