Proposing a novel molecular subtyping scheme for predicting distant recurrence-free survival in breast cancer post-neoadjuvant chemotherapy with close correlation to metabolism and senescence.

Background: High relapse rates remain a clinical challenge in the management of breast cancer (BC), with distant recurrence being a major driver of patient deterioration. To optimize the surveillance regimen for distant recurrence after neoadjuvant chemotherapy (NAC), we conducted a comprehensive analysis using bioinformatics and machine learning approaches. Materials and methods: Microarray data were retrieved from the GEO database, and differential expression analysis was performed with the R package 'Limma'. We used the Metascape tool for enrichment analyses, and 'WGCNA' was utilized to establish co-expression networks, selecting the soft threshold power with the 'pickSoftThreshold' algorithm. We integrated ten machine learning algorithms and 101 algorithm combinations to identify key genes associated with distant recurrence in BC. Unsupervised clustering was performed with the R package 'ConsensusCluster Plus'. To further screen the key gene signature of residual cancer burden (RCB), multiple knockdown studies were analyzed with the Genetic Perturbation Similarity Analysis (GPSA) database. Single-cell RNA sequencing (scRNA-seq) analysis was conducted through the Tumour Immune Single-cell Hub (TISCH) database, and the XSum algorithm was used to screen candidate small molecule drugs based on the Connectivity Map (CMAP) database. Molecular docking processes were conducted using Schrodinger software. GMT files containing gene sets associated with metabolism and senescence were obtained from GSEA MutSigDB database. The GSVA score for each gene set across diverse samples was computed using the ssGSEA function implemented in the GSVA package. Results: Our analysis, which combined Limma, WGCNA, and machine learning approaches, identified 16 RCB-relevant gene signatures influencing distant recurrence-free survival (DRFS) in BC patients following NAC. We then screened GATA3 as the key gene signature of high RCB index using GPSA analysis. A novel molecular subtyping scheme was developed to divide patients into two clusters (C1 and C2) with different distant recurrence risks. This molecular subtyping scheme was found to be closely associated with tumor metabolism and cellular senescence. Patients in cluster C2 had a poorer DRFS than those in cluster C1 (HR: 4.04; 95% CI: 2.60-6.29; log-rank test p < 0.0001). High GATA3 expression, high levels of resting mast cell infiltration, and a high proportion of estrogen receptor (ER)-positive patients contributed to better DRFS in cluster C1. We established a nomogram based on the N stage, RCB class, and molecular subtyping. The ROC curve for 5-year DRFS showed excellent predictive value (AUC=0.91, 95% CI: 0.95-0.86), with a C-index of 0.85 (95% CI: 0.81-0.90). Entinostat was identified as a potential small molecule compound to reverse high RCB after NAC. We also provided a comprehensive review of the EDCs exposures that potentially impact the effectiveness of NAC among BC patients. Conclusion: This study established a molecular classification scheme associated with tumor metabolism and cancer cell senescence to predict RCB and DRFS in BC patients after NAC. Furthermore, GATA3 was identified and validated as a key gene associated with BC recurrence.

Using Acupuncture for Adjunct Treatment for Cancer-Related Fatigue in Breast Cancer Patients Is Practicable. Reply to Hu et al. Is Acupuncture an Ideal Adjunctive Treatment for Cancer-Related Fatigue? Comment on "Choi et al. Acupuncture for Managing Cancer-Related Fatigue in Breast Cancer Patients: A Systematic Review and Meta-Analysis. Cancers 2022, 14, 4419".

We are grateful for the relevant comments by Hu et al. [...].

Acupuncture for Managing Cancer-Related Fatigue in Breast Cancer Patients: A Systematic Review and Meta-Analysis.

Breast cancer (BC) is the most common cancer in women and is a serious threat to women's health. Cancer-related fatigue (CRF) is a distressing symptom in BC patients during and after chemotherapy or radiation therapy that severely affects quality of life (QoL). AT is widely used for fatigue management. However, the effect of AT on CRF is still uncertain. This study aimed to evaluate the efficacy and safety of AT in the management of CRF in patients with BC. Eleven databases were searched through June 2022. Two researchers independently performed the database search, study selection, data extraction, and risk of bias assessment. Study selection was performed based on predefined Participants, Intervention, Comparators, Outcomes, Study design (PICOS) criteria, and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed when reporting the results. A meta-analysis was performed according to the Cochrane systematic review method using RevMan 5.3. A total of 12 studies including a total of 1084 participants were included. The results showed that AT had a beneficial effect compared with sham AT (n = 256, SMD = -0.26, 95% CI [-0.51, -0.01], p = 0.04, I2 = 0%) and a long-term effect on fatigue score (n = 209, MD = -0.32, 95% CI [-0.59, -0.04], p = 0.02, I2 = 0%). Meta-analysis showed that AT had a beneficial effect compared with usual care (UC) on fatigue scores (n = 238, SMD = -0.39, 95% CI [-0.66 to -0.12], p = 0.005, I2 = 0%). Of the 12 articles, 3 articles were judged as having a low risk of bias in all domains and hence were of high quality. No serious adverse effects were identified. AT is an effective and safe treatment for CRF, and AT is more effective than sham AT or UC or wait-list control (WLC). Nevertheless, the methodological quality of most of these studies was low, and the included studies/sample sizes were small, so the ability to derive decisive implications was limited. Further research is needed to confirm these findings.

Chimeric Oncolytic Adenovirus Armed Chemokine Rantes for Treatment of Breast Cancer.

The immunosuppressive state in the tumor microenvironment (TME) of breast cancer makes it difficult to treat with immunotherapy. Oncolytic viruses not only lyse tumor cells but also reshape the TME. Therefore, they can play a multi-mechanism synergistic effect with immunotherapy. In this study, an oncolytic adenovirus Ad5F11bSP-Rantes was constructed and used as a vector to express the chemokine Rantes. The objective of this study was to test the dual mechanisms of the oncolytic effect mediated by virus replication and the enhanced anticancer immune response mediated by Rantes chemotaxis of immune cells. It was found that Ad5F11bSP-Rantes has strong infectivity and effective killing activity against breast cancer cells. In the established triple negative breast cancer (TNBC) xenograft model in NCG mice whose immune system was humanized with human peripheral blood mononuclear cells (PBMCs), Ad5F11bSP-Rantes achieved 88.33% tumor inhibition rate. Rantes expression was high in mouse blood, a large number of CD3+ lymphocytes infiltrated in tumor tissues and E-cadherin was up-regulated in cancer cells, suggesting that Ad5F11bSP-Rantes altered the TME and induced a reversal of cancer cell epithelial-mesenchymal transition (EMT). In conclusion, oncolytic adenovirus can exert the oncolytic effect and the chemotactic effect of immune cells and realize the synergy of multiple anticancer effects. This strategy creates a candidate treatment for the optimization of breast cancer, especially TNBC, combination therapy.

Comment on "Efficacy of ginseng and its ingredients as adjuvants to chemotherapy in non-small cell lung cancer" by H. Zhu, H. Liu, J. H. Zhu, S. Y. Wang, S. S. Zhou, M. Kong, Q. Mao, F. Long, Z. J. Fang and S.-L. Li, Food Funct., 2021, 12, 2225.

This comment raises concerns about the article "Efficacy of ginseng and its ingredients as adjuvants to chemotherapy in non-small cell lung cancer".

Acupuncture and Moxibustion for Cancer-Related Fatigue: An Overview of Systematic Reviews and Meta-Analysis.

Although acupuncture (AT) is used in the treatment of CRF, the evidence from different systematic reviews (SRs) of AT has not yet been comprehensively evaluated. Moxibustion, which is a treatment method that is well established within Traditional East Asian Medicine, applies the heat of burning herbs towards or onto special points on the skin. Commonly, the herb Artemisia vulgaris, is used. It has been used for palliative cancer care, as well as for CRF. The aim of this overview was to evaluate the efficacy of AT and moxibustion in the management of CRF. Eleven databases were searched through for studies that were published from their dates of inception to February 2022. The study selection, the data extraction, and the assessment were performed independently by two researchers. The methodological and report quality were assessed by using the Assessment of Multiple Systematic Reviews-2 (AMSTAR-2) tool. The evidence quality was evaluated by using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. Fifteen SRs on AT (n = 10) and moxibustion (n = 5) treatments for CRF were included, and they include 169 randomized controlled trials and 14,392 participants. All of the SRs that were evaluated by the AMASTAR-2 had more than one deficiency, and so all of the SRs were rated as either low or critically low. For the GRADE, 18 outcomes were rated as very-low-quality evidence, 13 as low-quality evidence, 3 as moderate-quality evidence, and 0 as high-quality evidence. Most of the SRs reached the potential benefits of AT for CRF. No serious adverse effects were identified. In conclusion, the evidence suggests that, despite the advantages of AT in terms of the improvement in and the safety of the treatment of CRF, the methodological quality of most of these studies is low, which limits our ability to draw definitive meanings. Further research of high quality is needed in order to confirm these findings.

Triple-serotype chimeric oncolytic adenovirus exerts multiple synergistic mechanisms against solid tumors.

BACKGROUND: Oncolytic virotherapy has become an important branch of cancer immunotherapy. This study investigated the efficacy of an oncolytic adenovirus (OAV), OncoViron, with synergistic mechanisms in the treatment of multiple solid tumors. METHODS: An OAV, OncoViron, was constructed and investigated by cytological experiments and implanted tumor models of multiple solid tumor cell lines to certify its anticancer efficacy, the synergistic effects of viral oncolysis and transgene anticancer activity of OncoViron, as well as oncolytic virotherapy combined with immunotherapy, were also verified. RESULTS: The selective replication of OncoViron mediated high expression of anticancer factors, specifically targeted a variety of solid tumors and significantly inhibited cancer cell proliferation. On a variety of implanted solid tumor models in immunodeficient mice, immunocompetent mice, and humanized mice, OncoViron showed great anticancer effects on its own and in combination with programmed death 1 (PD-1) antibody and chimeric antigen receptor (CAR) T cells. Pathological examination, single-cell sequencing, and spatial transcriptome analysis of animal implanted tumor specimens confirmed that OncoViron significantly altered the gene expression profile of infected cancer cells, not only recruiting a large number of lymphocytes, natural killer cells, and mononuclear macrophages into tumor microenvironment (TME) and activated immune cells, especially T cells but also inducing M1 polarization of macrophages and promoting the release of more immune cytokines, thereby remodeling the TME for coordinating PD-1 antibody or CAR T therapy. CONCLUSIONS: The chimeric OncoViron is a novel broad-spectrum anticancer product with multiple mechanisms of synergistic and potentiated immunotherapy, creating a good opportunity for combined immunotherapy against solid tumors.

Effect of a multidisciplinary ward-based intervention on end-of-life care for general medicine patients.

OBJECTIVE: Providing good end-of-life (EOL) care for noncancer patients has been made a national priority in Singapore. A combined medical and nursing ward-based intervention known as the EOL care plan was piloted in a general medicine ward at our institution, aiming to guide key aspects of EOL care. The aim of this study is to assess the EOL care plan's effect on EOL care for general medicine patients. METHOD: We conducted a retrospective cohort study on inpatients who died in a general ward under the discipline "General Medicine" from May to October 2019. We collected data around symptom management, rationalization of care and communication with families. The primary analysis compared care received by patients who died in the pilot ward with that of a control group of patients who died in other wards. RESULTS: In total, 112 records were included in the analysis. Pain assessment was more common in the pilot ward compared with the control group (35.3% vs. 6.3%, p < 0.001), as were anti-psychotic prescriptions for delirium (64.7% vs. 24.4%, p = 0.001). Fewer patients received blood glucose monitoring in the last 48 h of life in the pilot ward (69.5% vs. 35.3%, p = 0.007). There were also less frequent parameters monitoring in the pilot ward (p < 0.004). SIGNIFICANCE OF RESULTS: The implementation of the EOL care plan was associated with process-level indicators of better EOL care, suggesting that it could have a significant positive impact when implemented on a wider scale.

Down-regulation of FOS-like antigen 1 enhances drug sensitivity in breast cancer.

OBJECTIVE: Multidrug resistance (MDR) to chemotherapeutic drugs is an important reason for clinical chemotherapy failure. So far, the relationship between FOS-like antigen1 (FOSL1) and chemotherapy sensitivity of breast cancer remains unclear. This study investigates the relationship between FOSL1 and chemotherapy sensitivity of breast cancer and its molecular mechanism. METHODS: Doxorubicin-resistant MCF-7/ADR breast cancer cells were transfected with NC (control) or FOSL1 siRNA and assayed for cell viability and relative colony number by MTT assay and colony formation, respectively. The expression level of FOSL1 was detected by immunohistochemistry (IHC). The relationship between FOSL1 and chemotherapy sensitivity was analyzed by a one-way of variance analysis and Pearson's chi-square test among a total of 50 patients with stage II and III breast cancer before and after they received epirubicin-based neoadjuvant chemotherapy (NCT) between 2012 and 2017. RESULTS: The expression of FOSL1 was increased in breast cancer tissues compared with normal breast tissues (P<0.05), and the expression of FOSL1 was decreased after NCT treatment compared with breast cancer tissues (or before NCT). This lower expression of FOSL1 was correlated with chemotherapy resistance or chemotherapy sensitivity (P<0.05). Moreover, the expression level of FOSL1 was markedly lower in NCT-sensitive patients than that of NCT-resistant patients (P<0.05). CONCLUSION: Down-regulation of FOSL1 potentiated chemotherapy sensitivity of breast cancer, and its lower expression attenuated chemotherapeutic drug resistance in human breast cancer cells. FOSL1 might be a drug target for predicting chemotherapy effect in breast cancer.

Oncolytic virotherapy armed with an engineered interfering lncRNA exhibits antitumor activity by blocking the epithelial mesenchymal transition in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) has special characteristics of significant aggressiveness, and strong potential for metastasis and recurrence; currently there are no targeted drugs for TNBC. Abnormal activation of epithelial-mesenchymal transition (EMT) plays an important role in these malignant behaviors of TNBC. In the crosstalk among the multiple EMT-associated signaling pathways, many miRNAs participate in regulating pathway activity, where they act as "traffic lights" at the intersection of these pathways. In this study, we used miRNA microarray technology to detect differentially expressed miRNAs related to EMT in TNBC, and we identified and verified 9 highly expressed oncogenic miRNAs (OncomiRs). High expression of these OncomiRs in clinical breast cancer tissues affected the prognosis of patients, and inhibition of their expression blocked EMT in TNBC cell lines and suppressed cancer cell proliferation and migration. We constructed an oncolytic adenovirus (AdSVP-lncRNAi9) armed with an artificially-designed interfering lncRNA (lncRNAi9), which exhibited an activity to block EMT in TNBC cells by disrupting the functions of multiple OncomiRs; the efficacy of such a treatment for TNBC was demonstrated in cytology and animal experiments. This research provides a new candidate oncolytic virotherapy for treating highly malignant refractory TNBC.

Extracellular vesicles-derived OncomiRs mediate communication between cancer cells and cancer-associated hepatic stellate cells in hepatocellular carcinoma microenvironment.

Tumor microenvironment (TME) is a critical determinant for hepatocellular carcinoma (HCC). Hepatic stellate cells (HSCs) are main interstitial cells in TME and play a vital role in early intrahepatic invasion and metastasis of HCC. The potential mechanism on the interactions between HSCs and HCC cells remains unclear. In this study, the effects of extracellular vesicles (EVs)-derived OncomiRs that mediate communication between HCC cells and cancer-associated hepatic stellate cells (caHSCs) and remold TME were investigated. The results found that the HCC cells-released EVs contained more various OncomiRs, which could activate HSCs (LX2 cells) and transform them to caHSCs, the caHSCs in turn exerted promotion effects on HCC cells through HSCs-released EVs. To further simulate the effects of OncomiRs in EVs on construction of pro-metastatic TME, a group of OncomiRs, miR-21, miR-221 and miR-151 was transfected into HCC cells and LX2 cells. These microRNAs in the EVs from OncomiRs-enhanced cells were demonstrated to have oncogenic effects on HCC cells by upregulating the activities of protein kinase B (AKT)/extracellular signal-regulated kinase (ERK) signal pathways. Equivalent results were also found in HCC xenografted tumor models. The findings suggested that the OncomiR secretion and transference by cancer cells-released EVs can mediate the communication between HCC cells and HSCs. HCC cells and caHSCs, as well as their secreted EVs, jointly construct a pro-metastatic TME suitable for invasion and metastasis of cancer cells, all these TME components form a positive feedback loop to promote HCC progression and metastasis.

MicroRNA-130a reduces drug resistance in breast cancer.

OBJECTIVE: Although the advent of chemotherapy has made some progress in the comprehensive treatment of breast cancer, drug resistance of tumor cells remains to be one of the main challenges for the treatment of breast cancers. Several microRNAs have been implicated in the resistant process, but the role of miR-130a in drug resistance in breast cancer remains unclear. The present study aims to investigate the role and mechanisms of miR-130a in drug resistance in breast cancer cells and tissues. PATIENTS AND METHODS: miR-130a mimics was used to up-regulate miR-130a expression in Doxorubicin-resistant MCF-7/Adr breast cancer cell line, followed by MTT assay and colony formation to determine cell viability and relative colony number. The relationship between the expression of miR-130a and drug resistance was detected by in situ hybridization in the formalin-fixed paraffin-embedded (FFPE) tissues from 50 breast cancer patients before and after Epirubicin-based neoadjuvant chemotherapy. RESULTS: Up-regulation of miR-130a level in MCF-7/Adr cells decreased the cell viability and colony number, and reversed Doxorubicin resistance of MCF-7/Adr cells. In breast cancer tissue from patients, the miR-130a level was lower before neoadjuvant chemotherapy than that after neoadjuvant chemotherapy (P < 0.05). Moreover, a significant increase in the expression of miR-130a was observed in breast tumor tissues from patients sensitive to neoadjuvant chemotherapy compared to the patients who were resistant to neoadjuvant chemotherapy (P < 0.05). CONCLUSION: We concluded that miR-130a might weaken drug resistance of human breast cancer cells, and act as an important factor in prediction of therapeutic responses in chemotherapy of breast cancer.

Primary epithelioid angiosarcoma of right hip joint: A case report and literature review.

RATIONALE: Epithelioid angiosarcoma (EA) is a rare, highly invasive tumor. The histopathological features of EA are not distinct and less reported in the literature, and most of the medical records are incomplete. PATIENT CONCERNS: A 61-year-old woman who came to the hospital because of pain in her right hip. This patient had had surgery for right hip tuberculosis 30 years ago. DIAGNOSES: The present study reports a case of primary EA of bone with aneurysmal bone cyst (ABC) that was diagnosed by 3 experienced pathologists. INTERVENTIONS: The patients had undergone 2 surgeries; however, an early recurrence of the tumor was caused the death of the patient. OUTCOMES: Ten EA cases from other literature were reviewed in this article; all the symptoms were found in different parts of bone, and the case data were relatively complete. The primary clinical features and nonspecific histopathological morphology of the disease were summarized from the 11 cases mentioned in the literature, and the main immunohistochemistry characteristics and diagnostic traps of EA were reviewed. LESSONS: Because the tumor has no characteristic diagnostic index in imaging and laboratory examination, the histopathologic features are not typical, especially in the case of obvious secondary lesions. It is easy to miss and misdiagnose. If possible, the diagnosis should be combined with immunohistochemical results.

Risk factors for lower extremity injuries in young badminton players / Fatores de risco para lesões de extremidade inferior em jovens jogadores de badminton

AIMS: Based on the limited evidence available about the intrinsic factors causing lower extremity injuries among Malaysian badminton players, this study was aimed to determine the relationship of demographic and physical characteristics to lower extremity injuries in young badminton players. METHODS: A cross-sectional study included badminton players between 14 and 24 years of age, categorized into case and control groups. Participants diagnosed with lower limb injuries were designated as cases, and those with no reported injuries were designated as controls. Personal information including demographic data, level of athlete and injury history was collected using a questionnaire. Independent t-test was used to analyze the differences between intrinsic characteristics in cases and controls. Pearson's χ2 was applied to evaluate the association between risk factors and general lower limb injuries, knee injuries and ankle injuries, with 95% confidence interval (CI). A p value of ≤0.05 was considered significant. RESULTS: A total of 106 young badminton players (83 males, 23 females) were recruited, of whom 42 participants were allocated as the case group, and 64 participants were allocated as the control group. A total of 60 lower extremity injuries were reported among the 42 players of the case group. The overall mean age of the sample was 18.7±5 years (minimum 14 years and maximum 24 years). Mean age of the participants in the case group was 16.92±2.99 years. The most common injuries reported were ankle joint injuries, followed by knee and hip injuries. Participants of the younger age group (14-19 years old) were found to have a higher risk for lower extremity injures compared to those of the older age group (20-24 years old) (odds ratio [OR], 3.39; 95%CI, 1.15-10.01; p=0.023). Increased true limb length discrepancy was identified among the participants with lower extremity injuries (OR, 4.57, 95%CI, 1.2-17.24; p=0.016) and this discrepancy was strongly associated with ankle injuries (OR, 7.25; 95%CI, 1.85-28.57; p=0.002). There was no significant relationship between lower extremity injuries and gender, limb dominance or Q-angle. CONCLUSIONS: Lower extremity injuries in young badminton players were predominantly located in ankle and knee joints. Younger age and increase in true limb length discrepancy were identified as risk factors for lower extremity injuries in the study sample.

OBJETIVOS: Com base na limitada evidência disponível sobre os fatores intrínsecos que causam lesões de extremidade inferior entre os jogadores de badminton da Malásia, este estudo teve como objetivo avaliar as relações de características demográficas e físicas com lesões nas extremidades inferiores em jovens jogadores de badminton. MÉTODOS: Um estudo transversal incluiu jogadores de badminton entre 14 e 24 anos de idade, categorizados em grupos de casos e controles. Os participantes diagnosticados com lesões dos membros inferiores foram designados como casos, e aqueles sem lesões relatadas foram designados como controles. Informações pessoais, incluindo dados demográficos, nível de atleta e histórico de lesões foram coletadas usando um questionário. O teste t independente foi utilizado para analisar as diferenças entre características intrínsecas em casos e controles. O χ2 de Pearson foi aplicado para avaliar a associação entre fatores de risco e lesões de membro inferior em geral, lesões no joelho e lesões no tornozelo, com intervalo de confiança (IC) de 95%. Um valor de p ≤0,05 foi considerado como significativo. RESULTADOS: Um total de 106 jovens jogadores de badminton (83 do gênero masculino e 23 do gênero feminino) foram recrutados, dos quais 42 participantes foram alocados no grupo de casos e 64 participantes foram alocados no grupo controle. Um total de 60 lesões nas extremidades inferiores foram relatadas entre os 42 jogadores do grupo de casos. A média de idade da amostra total foi de 18,7±5 anos (mínima 14 anos e máxima 24 anos). A média de idade dos participantes no grupo de casos foi de 16,92±2,99 anos. As lesões mais comumente relatadas foram as localizadas na articulação do tornozelo, seguidas das lesões de joelho e de quadril. Os participantes da faixa etária mais jovem (14-19 anos de idade) apresentaram maior risco de lesões nas extremidades inferiores em comparação com os de faixa etária mais velha (20-24 anos de idade) (odds ratio [OR], 3.39; 95% CI, 1,15-10,01; p=0,023). Maior discrepância verdadeira no comprimento dos membros inferiores foi identificada entre os participantes com lesões nas extremidades inferiores (OR, 4,57, IC 95%, 1,2-17,24; p=0,016) e esta discrepância foi fortemente associada a lesões no tornozelo (OR, 7,25; IC95%, 1,85 -28,57; p=0,002). Não houve relações significativas entre lesões das extremidades inferiores e gênero, dominância dos membros ou ângulo Q. CONCLUSÕES: As lesões das extremidades inferiores em jovens jogadores de badminton foram predominantemente localizadas nas articulações do tornozelo e do joelho. A faixa etária mais jovem e o aumento da discrepância verdadeira no comprimento dos membros foram identificados como fatores de risco para lesões das extremidades inferiores na amostra estudada.

Expression of and correlation between BCL6 and ZEB family members in patients with breast cancer.

B-cell lymphoma 6 (BCL6), a proto-oncogene, is an evolutionarily conserved zinc finger protein that functions as a transcriptional repressor. BCL6 is the master regulator of B-lymphocyte development, and it has been reported that BCL6 may serve an important role in breast cancer progression. The aim of the present study was to investigate the expression of BCL6, zinc finger E-box-binding homeobox (ZEB)1 and ZEB2 and their associations in breast cancer. The mRNA and protein expression of BCL6, ZEB1 and ZEB2 was assessed using in situ hybridization and immunohistochemistry, respectively, in 228 patients with breast cancer and 80 patients with benign breast disease. In addition, the association between BCL6, ZEB1 and ZEB2 expression and the clinicopathological characteristics and survival of patients with breast cancer were analyzed. The mRNA and protein expression of BCL6, ZEB1 and ZEB2 were significantly higher in breast cancer tissues compared with benign breast disease tissues (P<0.05). The expression of BCL6, ZEB1 and ZEB2 were significantly positively correlated with tumor size, lymph node metastasis and a higher tumor stage (P<0.05). Furthermore, patients with BCL6, ZEB1 and ZEB2 protein-positive primary tumors had significantly lower overall survival (P=0.001, 0.002 and 0.001, respectively) and relapse-free survival (P=0.002, 0.001 and 0.003, respectively) rates. The mRNA expressions of ZEB1 (rs=0.326, P<0.001) and ZEB2 (rs=0.382, P<0.001) were significantly positively correlated with BCL6 mRNA expression, and the protein expressions of ZEB1 ((rs=0.449, P<0.001) and ZEB2 (rs=0.669, P<0.001) were significantly positively correlated with BCL6 protein expression. These results suggest that BCL6, ZEB1 and ZEB2 are potential biomarkers for the invasion, metastasis and prognosis of breast cancer, and that BCL6 may be a regulator of the ZEB family.

MicroRNAs regulate the epithelial-mesenchymal transition and influence breast cancer invasion and metastasis.

MicroRNAs are small RNA molecules that play a major role in the post-transcriptional regulation of genes and influence the development, differentiation, proliferation, and apoptosis of cells and the development and progression of tumors. The epithelial-mesenchymal transition is a process by which epithelial cells morphologically transform into cells with a mesenchymal phenotype. The epithelial-mesenchymal transition plays a highly important role in tumor invasion and metastasis. Increasing evidence indicates that microRNAs are tightly associated with epithelial-mesenchymal transition regulation in tumor cells. In breast cancer, various microRNA molecules have been identified as epithelial-mesenchymal transition inducers or inhibitors, which, through different mechanisms and signaling pathways, participate in the regulation of breast cancer invasion and metastasis among various biological behaviors. The epithelial-mesenchymal transition-related microRNAs in breast cancer provide valuable molecules for researching cell invasion and metastasis, and they also provide candidate targets that may be significant for the targeted therapy of breast cancer.

Autoregulatory feedback loop of EZH2/miR-200c/E2F3 as a driving force for prostate cancer development.

The histone methyltransferase enhancer of zeste homolog 2 (EZH2) has recently attracted considerable attention because of its dysregulation in prostate cancer (PCa) and its important function in PCa development. To date, little is known about the underlying cellular function and regulatory networks of EZH2 in PCa. This study aims to determine whether or not the autoregulatory feedback loop of EZH2/miR-200c/E2F3 serves key functions in PCa development. Bioinformatics and integrative analytical approaches were employed to identify the relationships of EZH2 to specific cancer-related gene sets. Results indicated that the enrichment of gene sets about cell cycle progression was associated with EZH2 expression. The depletion of EZH2 in cell experiments inhibited PCa cell growth and blocked cell cycle accompanying the downregulation of E2F3 expression. Furthermore, miR-200c served as an important mediator between EZH2 and E2F3. Compared with scrambled control cells, sh-EZH2 cells showed lower H3K27me3 expression and higher miR-200c expression. Western blot and luciferase reporter assays showed that miR-200c inversely modulated E2F3 by directly targeting the binding site within 3'UTR. Moreover, decreased miR-200c expression largely abrogated the effect of sh-EZH2 on E2F3 expression and E2F3-induced cell cycle progression. EZH2 was positively regulated by E2F3 at the transcriptional level. Immunohistochemistry and in situ hybridization revealed a significant correlation among EZH2, miR-200c, and E2F3 expression in human PCa tissues. In conclusion, the autoregulatory feedback loop of EZH2/miR-200c/E2F3 served an important function in PCa development. Targeting this aberrantly activated feedback loop may provide a new therapeutic strategy against PCa.

Hypoxic damage to the periventricular white matter in neonatal brain: role of vascular endothelial growth factor, nitric oxide and excitotoxicity.

The present study examined factors that may be involved in the development of hypoxic periventricular white matter damage in the neonatal brain. Wistar rats (1-day old) were subjected to hypoxia and the periventricular white matter (corpus callosum) was examined for the mRNA and protein expression of hypoxia-inducible factor-1alpha (HIF-1alpha), endothelial, neuronal and inducible nitric oxide synthase (eNOS, nNOS and iNOS), vascular endothelial growth factor (VEGF) and N-methyl-D-aspartate receptor subunit 1 (NMDAR1) between 3 h and 14 days after hypoxic exposure by real-time RT-PCR, western blotting and immunohistochemistry. Up-regulated mRNA and protein expression of HIF-1alpha, VEGF, NMDAR1, eNOS, nNOS and iNOS in corpus callosum was observed in response to hypoxia. NMDAR1 and iNOS expression was found in the activated microglial cells, whereas VEGF was localized to astrocytes. An enzyme immunoassay showed that the VEGF concentration in corpus callosum was significantly higher up to 7 days after hypoxic exposure. NO levels, measured by colorimetric assay, were also significantly higher in hypoxic rats up to 14 days after hypoxic exposure as compared with the controls. A large number of axons undergoing degeneration were observed between 3 h and 7 days after the hypoxic exposure at electron-microscopic level. Our findings point towards the involvement of excitotoxicity, VEGF and NO in periventricular white matter damage in response to hypoxia.