Objective Preeclampsia is a major cause of maternal, fetal, and neonatal mortality. It is necessary to improve early detection of preeclampsia. This study aims to explore a plasma exosome miRNA as a potential biomarker for preeclampsia. Methods In this study, we adopted a common extraction method for exosomes - SEC combined with ultrafiltration and the ExoQuick commercial kit method - to extract exosomes from the plasma of patients with preeclampsia and healthy pregnant women, and screen for suitable endogenous reference genes. By applying bioinformatics, we identified differentially expressed miRNAs in the plasma exosomes of patients with preeclampsia and healthy pregnant women, and further confirmed their significance in the clinical diagnosis of preeclampsia. Results By using SEC combined with ultrafiltration (SEC group) and ExoQuick (ExoQuick group) commercial kit exosome separation, we extracted exosomes from the plasma of the preeclampsia group and the normal pregnant women group. MiR-30a and miR-30e-5p was verified as the optimal endogenous reference gene. We screened out 4 miRNAs (miR-122-5p, miR-126-3p, miR-144-3p and miR-451a) from 68 differentially expressed miRNAs for verification. Eventually, it was found that among the exosomes extracted by the two exosome isolation methods, only miR-122-5p was stably upregulated in patients with preeclampsia independently of the exosome extraction methods, which may have potential value for the diagnosis and treatment of preeclampsia. Conclusion In this study, we found that miR-122-5p maintained stable differences in plasma exosomes of patients with preeclampsia under different exosome extraction methods, thereby providing clues for biomarkers for the early diagnosis of preeclampsia.

Background: Breast cancer (BC) is a prevalent malignancy among women, and numerous investigations have reported that platelet aggregation may play a role in BC progression. Thus, identifying new targets for BC is essential. In this regard, we focused on nucleolar protein 6 (NOL6), located on chromosome 9p13, which is implicated in tumor development. Objective: To investigate NOL6 expression in BC, examine its role in platelet aggregation and angiogenesis, and elucidate the underlying mechanisms. Methods: Bioinformatic analyses, immunoblotting, and quantitative real-time polymerase chain reaction (qPCR) were performed to assess NOL6 expression in BC. Cell counting kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assays were conducted to determine the impact of NOL6 on BC cell proliferation. Immunostaining, enzyme-linked immunosorbent assay (ELISA), and flow cytometry (FCM) assays were utilized to analyze the effects of NOL6 on platelet aggregation. Tube formation and transwell assays were performed to examine angiogenesis and invasion, immunoblot assays were used to confirm the underlying mechanisms, and tumor growth assays in mice were conducted to validate the findings in vivo. Results: NOL6 was found to be highly expressed in BC and was associated with patient prognosis, platelet aggregation, and angiogenesis. Its knockdown inhibited BC cell proliferation and reduced platelet aggregation induced by BC cells. Additionally, NOL6 depletion impaired angiogenesis and migration of BC cells. In vivo studies confirmed that NOL6 promotes tumor growth. Mechanistically, NOL6 enhances the Twisted spiral transcription factor 1 (Twist1)/galectin-3 axis, contributing to BC progression. Conclusions: NOL6 can promote tumor progression by facilitating platelet aggregation and angiogenesis in BC cells through the Twist1/galectin-3 axis.

We previously screened a peptide PDBAG1 that remarkably inhibited triple-negative breast cancer, and found that its target was C1QBP. Recently, C1QBP has been reported as a potential tumor marker in ovarian cancer, which of the mortality rate ranks first among malignant tumors of the female reproductive tract. However, it is unclear whether and how PDBAG1 plays a regulatory role in ovarian cancer. Here, we first found that PDBAG1 definitely inhibited the growth and metastasis of ovarian cancer in vitro and in vivo. PDBAG1 downregulated the protein level of C1QBP and damaged mitochondria in ovarian cancer. Furthermore, we analyzed the overall impact of PDBAG1 on ovarian cancer cells through transcriptomics, and found that KRAS, inflammation and stress-related signals were dramatically activated. The accuracy of the transcriptome sequencing results was also subsequently verified. Moreover, we combined the inhibitors of the classic downstream MAPK signaling pathway of KRAS and the integrated stress inhibitor with PDBAG1, and found that the p38 MAPK inhibitor, Adezmapimod, significantly enhanced the inhibitory effect of PDBAG1 on ovarian cancer and inhibited the upregulation of the crucial stress response transcription factor ATF4 caused by PDBAG1. Collectively, our research results revealed the function and mechanism of the peptide PDBAG1 in ovarian cancer, providing new insights into clinical drug development for ovarian cancer.

AbstractC1QBP exhibits heightened expression across a spectrum of tumours, thereby fostering their proliferation and metastasis, rendering it a pivotal therapeutic target. Nevertheless, to date, no pharmacological agents capable of directly targeting and inducing the degradation of C1QBP have been identified. In this study, we have unveiled a new peptide, PDBAG1, derived from the precursor protein GPD1, employing a peptidomics‐based drug screening strategy. PDBAG1 has demonstrated substantial efficacy in suppressing triple‐negative breast cancer (TNBC) both in vitro and in vivo. Its mechanism of action involves mitochondrial impairment and the inhibition of oxidative phosphorylation (OXPHOS), achieved through direct binding to C1QBP, thereby promoting its ubiquitin‐dependent degradation. Concomitantly, due to metabolic adaptability, we have observed an up‐regulation of glycolysis to compensate for OXPHOS inhibition. We observed an aberrant phenomenon wherein the hypoxia signalling pathway in tumour cells exhibited significant activation under normoxic conditions following PDBAG1 treatment. Through size‐exclusion chromatography (SEC) and isothermal titration calorimetry (ITC) assays, we have validated that PDBAG1 is capable of binding C1QBP with a Kd value of 334 nM. Furthermore, PDBAG1 inhibits homologous recombination repair proteins and facilitates synergism with poly‐ADP‐ribose polymerase inhibitors in cancer therapy. This underscores that PDBAG1 ultimately induces insurmountable survival stress through multiple mechanisms while concurrently engendering therapeutic vulnerabilities specific to TNBC.Key points The newly discovered peptide PDBAG1 is the first small molecule substance found to directly target and degrade C1QBP, demonstrating significant tumour inhibitory effects and therapeutic potential.

AbstractBackgroundThe gastrointestinal tract contains a wide range of microorganisms that have evolved alongside the immune system of the host. The intestinal mucosa maintains balance within the intestines by utilizing the mucosal immune system, which is controlled by the complex gut mucosal immune network.ObjectiveThis review aims to comprehensively introduce current knowledge of the gut mucosal immune system, focusing on its interaction with commensal bacteria.ResultsThe gut mucosal immune network includes gut‐associated lymphoid tissue, mucosal immune cells, cytokines, and chemokines. The connection between microbiota and the immune system occurs through the engagement of bacterial components with pattern recognition receptors found in the intestinal epithelium and antigen‐presenting cells. This interaction leads to the activation of both innate and adaptive immune responses. The interaction between the microbial community and the host is vital for maintaining the balance and health of the host's mucosal system.ConclusionThe gut mucosal immune network maintains a delicate equilibrium between active immunity, which defends against infections and damaging non‐self antigens, and immunological tolerance, which allows for the presence of commensal microbiota and dietary antigens. This balance is crucial for the maintenance of intestinal health and homeostasis. Disturbance of gut homeostasis leads to enduring or severe gastrointestinal ailments, such as colorectal cancer and inflammatory bowel disease. Utilizing these factors can aid in the development of cutting‐edge mucosal vaccines that have the ability to elicit strong protective immune responses at the primary sites of pathogen invasion.

Abstract Background Preeclampsia (PE) is a serious pregnancy complication associated with impaired trophoblast function. Integrin β3 (ITGB3) is a cell adhesion molecule that plays a role in cell movement. The objective of this study was to identify the biological function and expression level of ITGB3 in PE. Methods Cell proliferation, migration, invasion, adhesion, and apoptosis were estimated by CCK8 assay, transwell, scratch assays, and flow cytometry, respectively. The expression levels of ITGB3 were determined by qRT-PCR, western blot, and immunohistochemistry (IHC). Co-immunoprecipitation and Alphafold-Multimer protein complex structure prediction software were employed to identify the molecules that interact with ITGB3. Results Cell functional experiments conducted on HTR8/SVneo cells demonstrated that ITGB3 significantly enhanced proliferation, migration, invasion, and adhesion, while simultaneously inhibiting apoptosis. Relative ITGB3 expression levels were observed to be lower in PE placental tissue than in normal tissue and similarly reduced in hypoxic HTR8/SVneo cells. RNA-sequencing data from PE placental samples in the GEO database were analyzed to identify differentially expressed genes associated with the disease. We identified a total of 1460 mRNAs that were significantly differentially expressed in PE patients. Specifically, 798 mRNAs were significantly upregulated, and 662 mRNAs were significantly downregulated. Notably, the ITGB3 exhibited a pronounced down-regulation among the differential expression mRNA. Conclusions This study suggested that ITGB3 plays an important role in promoting the proliferative, migratory, invasive, and adhesive capabilities of trophoblast cells. These findings may facilitate a more in-depth understanding of the molecular mechanisms that promote PE progression.

The intestinal mucosal immune system is a specialized immune system that interacts closely with commensal microorganisms. The intestinal mucosal tissues are characterized by the way they generate immune responses to pathogens. The symbiotic flora in turn regulates the host’s mucosal immunity, maintaining a dynamic balance. The disruption of the equilibrium between the two can result in intestinal disorders. This paper summarises in depth the relationships between the two and serves as a resource for future research.

BackgroundLong-non-coding RNA PVT1 (lncRNA PVT1) can be used as an oncogenic regulatory non-coding RNA (ncRNA) for many cancers. However, its function and mechanism in breast cancer (BRCA) are still not clearly elucidated.ObjectiveWe attempt to explain the mechanism of PVT1's role in breast cancer from different perspectives.MethodsWe analyzed the expression of PVT1 and its correlation with the breast cancer related clinical data in the The Cancer Genome Atlas (TCGA) database. We used PVT1 overexpression and knockdown lentivirus to infect breast cancer MDA-MB-231 cell line for cell function verification, in vitro using CCK-8 to measure proliferation, flow cytometry to measure apoptosis, transwell test to measure invasion and migration ability, detecting cell extracellular acidification rate (ECAR) to assess glycolysis metabolism and explore the biological functions of PVT1 in breast cancer cells. Transcriptome sequencing was used to analyze the changes of related genes in cells after overexpression of PVT1. In vivo we used a xenograft model to study the effect of PVT1 on breast cancer.ResultsPVT1 was up-regulated in breast cancer tissues and was positively correlated with the clinical stage of breast cancer patients. Overexpression of PVT1 in vitro promoted cell proliferation, migration and invasion, and promoted tumor growth in vivo. Knockdown of PVT1 led to the opposite biological consequence. Further bioinformatics analysis showed that PVT1 changes the glycolysis metabolism of tumors through regulation of glycolysis-related genes. In addition, the expression of miR-145-5p is negatively correlated with PVT1. We consider the possibility of PVT1 promoting cell proliferation and metastasis by regulating the aerobic glucose metabolism in breast cancer cells through sponging the miR-145-5p.ConclusionOur results reveal a potential pathway for competing endogenous RNA to regulate breast cancer glucose metabolism. PVT1 regulates glycolysis related genes expression by competitively binding to endogenous miR-145-5p in breast cancer cells to change the metabolic phenotype. This may Provide new ideas for precise molecular therapy targets for breast cancer.

Background: The programmed cell death 1 (PD-1) receptor is an immune checkpoint molecule that induces immune tolerance and mediates the immune escape of tumor cells. It is mainly expressed in immune cells such as T cells, B cells and monocytes. In recent years, studies have shown that tumor cell-intrinsic PD-1 plays different roles in the development of melanoma, Liver cancer and lung cancer. However, the expression and function of PD-1 in colon cancer cells has not been reported. Our previous studies have found that Candida tropicalis (C. tropicalis) can promote CRC tumor growth and chemotherapy resistance to oxaliplatin by regulating mismatch repair system. Whether C. tropicalis participates in the progression of CRC and immunotherapy resistance through regulating the tumor cell-intrinsic PD-1 remains to be further elucidated. Methods & Results: In this study, we first found that high concentrations of C. tropicalis promote tumor growth in cell cultures and xenografts. In addition, we proved that colon cancer cell lines express PD-1 receptors. Knockdown of PD-1 enhanced SW480 viability in-vitro, while overexpression of PD-1 diminished cell viability. Moreover, blocking antibody against PD-1 promotes tumor growth both in SW480 cells and mice CRC xenografts in an adaptive immune-independent manner. We also demonstrated that high concentrations of C. tropicalis can down-regulate tumor cell-intrinsic PD-1 expression in colon cancer cells. CRC cell growth induced by C. tropicalis is partially offset in the presence of PD-1 overexpression. This shows that C. tropicalis promotes CRC progression via controlling the expression of tumor cell-intrinsic PD-1. Mechanistically, we found that C. tropicalis modulates the expression of PD-1 via increasing the autophagy traffic in colon cancer cells. Combining autophagy inhibitor with C. tropicalis treatment partly blocked the CRC tumor growth and reversed the downregulation of PD-1. Conclusion: This study shows that PD-1 is a tumor suppressor in CRC. C. tropicalis can down-regulate tumor cell-intrinsic PD-1 expression via enhancing tumor cells autophagy levels to promote CRC progression. It may provide a new idea and mechanism for answering why the immune monoclonal antibody treatment is ineffective in cancer patients.

Background: PANoptosis is a novel form of programmed cell death (PCD) found in 2019 that is regulated by the PANoptosome. PANoptosis combines essential features of pyroptosis, apoptosis, and necroptosis, forming a "death triangle" of cells. While apoptosis, pyroptosis, and necroptosis have been extensively studied for their roles in human inflammatory diseases and many other clinical conditions, historically they were considered as independent processes. However, emerging evidence indicates that these PCDs exhibit cross talk and interactions, resulting in the development of the concept of PANoptosis.Methods: In this review, we offer a concise summary of the fundamental mechanisms of apoptosis, pyroptosis, and necroptosis. We subsequently introduce the notion of PANoptosis and detail the assembly mechanism of the PANoptosome complex which is responsible for inducing cell death. We also describe some regulatory networks of PANoptosis.Results: PANoptosis now has been associated with various human diseases including cancer. Although the exact function of PANoptosis in each tumor is not fully understood, it represents a prospective avenue for cancer therapy, offering promise for advancements in cancer therapy.Conclusions: In the future, in-depth study of PANoptosis will continue to help us in understanding the fundamental processes underlying cell death and provide scientific support for cancer research.

BACKGROUND Hepatocellular carcinoma (HCC) is among the commonest cancer and is high in incidence. Besides, glycolysis has been proven to be a promoter in cancer progression. But the research related to glycolysis concentrates on tumor cells, and few are about macrophages. Dectin3 is a C-type Lectin receptor (CLR), expressed by myeloid lineage cells such as monocytes/macrophages, which can recognize pathogens and modulate immunity. We speculate that Dectin3 is involved in HCC by regulating the glycolysis in macrophages, which is meaningful. METHODS Wild-type (WT) mice and Dectin3-/- mice were used to establish a mouse model of HCC and the progression of HCC was evaluated. Primary tumor associated macrophages (TAMs) were isolated from tumor tissues and the level of glycolysis was assessed. WT and Dectin3-/- tumor-bearing mice were treated with glycolysis inhibitors and the tumor progression was assessed. Culture supernatant derived from H22 cells was used to stimulate bone-marrow-derived macrophages (BMDMs). The level of glycolysis in BMDMs was subsequently detected. H22 cells and BMDMs were co-cultured and then the proliferation and apoptosis of H22 cells were evaluated. RESULTS Compared with WT mice, tumor volume of Dectin3-/- mice increased, and the proportion of macrophages in tumor tissues increased, while the proportions of CD4+ and CD8+T cells decreased. Besides, the splenomegaly of Dectin3-/- mice was more serious. The level of glycolysis in macrophages of Dectin3-/- tumor-bearing mice was significantly up-regulated. After glycolysis inhibitor treatment, cancer progression of Dectin3-/- tumor-bearing mice slowed down, and the difference between WT mice and Dectin3-/- mice was significantly down-regulated. In addition, Dectin3 deficiency macrophages significantly promoted H22 cell proliferation and inhibited H22 cell apoptosis. CONCLUSION Dectin3 can protect against HCC. Dectin3 contributes to the apoptosis of tumor cells and inhibits the proliferation of tumor cells by regulating the glycolysis of macrophages.

Abstract Background Accumulating evidence implicates that gut fungi are associated with the pathogenesis of colorectal cancer (CRC). Our previous study has revealed that Candida tropicalis (C. tropicalis) promotes colorectal tumorigenesis by enhancing immunosuppressive function of myeloid-derived suppressor cells (MDSCs) and increasing accumulation of MDSCs, but the underlying mechanisms remain unestablished. Methods Bone marrow–derived MDSCs were stimulated with C. tropicalis. RNA-sequencing analysis was performed to screen the differentially expressed genes. Quantitative real-time PCR and western blot were used to measure the expression of related proteins. Co-culture assay of MDSCs and CD8+ T cells was used to determine the immunosuppressive ability of MDSCs. Metabolomic analysis was conducted to detect metabolic reprogramming of MDSCs. Aerobic glycolysis of MDSCs was assessed by extracellular acidification rate (ECAR), glucose consumption and lactate production. A CAC mouse model was induced by AOM and DSS to determine the therapeutic action of TEPP-46. IHC and immunofluorescence were performed to examine the expression of PKM2, PKM2 (p-Y105) and iNOS in human CRC-infiltrated MDSCs. Results C. tropicalis facilitates immunosuppressive function of MDSCs by increasing the expression of iNOS, COX2 and NOX2, production of nitric oxide (NO) and reactive oxygen species (ROS). Mechanistically, C. tropicalis facilitates the immunosuppressive function of MDSCs through the C-type lectin receptors Dectin-3 and Syk. C. tropicalis-enhanced immunosuppressive function of MDSCs is further dependent on aerobic glycolysis. On the one hand, NO produced by MDSCs enhanced aerobic glycolysis in a positive feedback manner. On the other hand, C. tropicalis promotes p-Syk binding to PKM2, which results in PKM2 Tyr105 phosphorylation and PKM2 nuclear translocation in MDSCs. Nuclear PKM2 interacts with HIF-1α and subsequently upregulates the expression of HIF-1α target genes encoding glycolytic enzymes, GLUT1, HK2, PKM2, LDHA and PDK1, which are required for the C. tropicalis-induced aerobic glycolysis of MDSCs. Blockade of PKM2 nuclear translocation attenuates C. tropicalis-mediated colorectal tumorigenesis. The high expression of PKM2, PKM2 (p-Y105) and iNOS in CRC-infiltrated MDSCs correlates with the development of human CRC. Conclusion C. tropicalis enhances immunosuppressive function of MDSCs via Syk-PKM2-HIF-1α-glycolysis signaling axis, which drives CRC. Therefore, we identify the Syk-PKM2-HIF-1α-glycolysis signaling axis as a potential therapeutic target for CRC.

Analyzing single-cell phenotypes is increasingly required in biomedical studies, for non-genetic understanding of cellular activities and the biological significance of rare cell subpopulations. However, as compared to the genotypic analysis, single-cell phenotype analysis is technically more challenging. Herein, a tractable method that allows quantitative phenotyping of single cell is developed in this work, termed as the aptamer-mounted nest-PCR (Apt-nPCR). In specific, only two rounds of PCR reactions are required to complete the analysis, where aptamers (short oligonucleotides that bind to specific target molecules) are used as the recognition elements to bind antigens and also as the templates of nPCR for multiplexed and quantitative detection. So, quantitative information of these target antigens can be revealed by quantitative PCR analysis of these aptamers, which can thus be used to interpret cell phenotypes in a quantitative-to-qualitative way. By addressing two technical issues that are involved in single-cell phenotype analysis─multiplexed detection plus high sensitivity, we have shown the availability of this method for single-cell phenotyping. Therefore, the Apt-nPCR method may represent a tractable method to facilitate the single-cell phenotype analysis, which can be used as a complementary method against these single-cell genotyping methods in our daily research.

Breast cancer is one of the most common malignant tumors in women worldwide. Circular RNA (circRNA) is a class of structurally stable non-coding RNA with a covalently closed circular structure. In recent years, with the development of high-throughput RNA sequencing, many circRNAs have been discovered and have proven to be clinically significant in the development and progression of breast cancer. Importantly, several regulators of circRNA biogenesis have been discovered. Here, we systematically summarize recent progress regarding the network of regulation governing the biogenesis, degradation, and distribution of circRNAs, and we comprehensively analyze the functions, mechanisms, and clinical significance of circRNA in breast cancer.

Background. Breast cancer is one of the most commonly diagnosed cancers all over the world, and it is now the leading cause of cancer death among females. The aim of this study was to find DEGs (differentially expressed genes) which can predict poor prognosis in breast cancer and be effective targets for breast cancer patients via bioinformatical analysis. Methods. GSE86374, GSE5364, and GSE70947 were chosen from the GEO database. DEGs between breast cancer tissues and normal breast tissues were picked out by GEO2R and Venn diagram software. Then, DAVID (Database for Annotation, Visualization, and Integrated Discovery) was used to analyze these DEGs in gene ontology (GO) including molecular function (MF), cellular component (CC), and biological process (BP) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway. Next, STRING (Search Tool for the Retrieval of Interacting Genes) was used to investigate potential protein-protein interaction (PPI) relationships among DEGs and these DEGs were analyzed by Molecular Complex Detection (MCODE) in Cytoscape. After that, UALCAN, GEPIA (gene expression profiling interactive analysis), and KM (Kaplan–Meier plotter) were used for the prognostic information and core genes were qualified. Results. There were 96 upregulated genes and 98 downregulated genes in this study. 55 upregulated genes were selected as hub genes in the PPI network. For validation in UALCAN, GEPIA, and KM, 5 core genes (KIF4A, RACGAP1, CKS2, SHCBP1, and HMMR) were found to highly expressed in breast cancer tissues with poor prognosis. They differentially expressed between different subclasses of breast cancer. Conclusion. These five genes (KIF4A, RACGAP1, CKS2, SHCBP1, and HMMR) could be potential targets for therapy in breast cancer and prediction of prognosis on the basis of bioinformatical analysis.

Triple-negative breast cancer (TNBC) is often treated with anthracyclines (e.g., epirubicin or doxorubicin), but very little is known about anthracycline resistance, especially epirubicin resistance in TNBC. To identify novel long noncoding RNAs (lncRNAs) involved in epirubicin resistance in TNBC, we established a new TNBC MDA-MB-231 cell line that was resistant to epirubicin (Epi-R). A total of 12 differentially expressed lncRNAs were identified using RNA sequencing analysis of Epi-R cells. Among these lncRNAs, we found a novel intronic lncRNA, lnc005620, was highly expressed in Epi-R cells and human TNBC tissues. Further gain- and loss-of-function studies demonstrated that lnc005620 played an oncogenic role and partially abrogated the effects of epirubicin on TNBC cells. Using iTRAQ proteomics analysis, we found that three members of the integrin family, integrin β4, integrin β1 and integrin α6, were all upregulated in Epi-R MDA-MB-231 cells. Integrin β1, encoded by the ITGB1 gene, was validated to be a downstream target of lnc005620 in Epi-R MDA-MB-231 cells. Our study demonstrates that novel lnc005620 promotes TNBC progression and chemoresistance to epirubicin via integrin β1 both in vitro and in vivo and provides a promising therapeutic target for TNBC patients in terms of enhancing the benefits of epirubicin treatment.

Breast cancer is one of the most common malignant tumors in women and is the second leading cause of cancer deaths among women. The tumorigenesis and progression of breast cancer are not well understood. The existing researches have indicated that non-coding RNAs, which mainly include long non-coding RNA (lncRNA) and microRNA (miRNA), have gradually become important regulators of breast cancer. We aimed to screen the differential expression of miRNA and lncRNA in the different breast cancer stages and identify the key non-coding RNA using TCGA data. Based on series test of cluster (STC) analysis, bioinformatics analysis, and negatively correlated relationships, 122 lncRNAs, 67 miRNAs, and 119 mRNAs were selected to construct the regulatory network of lncRNA and miRNA. It was shown that the miR-93/20b/106a/106b family was at the center of the regulatory network. Furthermore, 6 miRNAs, 10 lncRNAs, and 15 mRNAs were significantly associated with the overall survival (OS, log-rank P < 0.05) of patients with breast cancer. Overexpressed miR-93 in MCF-7 breast cancer cells was associated with suppressed expression of multiple lncRNAs, and these downregulated lncRNAs (MESTIT1, LOC100128164, and DNMBP-AS1) were significantly associated with poor overall survival in breast cancer patients. Therefore, the miR-93/20b/106a/106b family at the core of the regulatory network discovered by our analysis above may be extremely important for the regulation of lncRNA expression and the progression of breast cancer. The identified key miRNA and lncRNA will enhance the understanding of molecular mechanisms of breast cancer progression. Targeting these key non-coding RNA may provide new therapeutic strategies for breast cancer treatment and may prevent the progression of breast cancer from an early stage to an advanced stage.

Human epidermal growth factor receptor 2 (HER2)-positive is a particularly aggressive type of the breast cancer. Because of the evidence has revealed that accurate HER2 status detection is crucial for prognosis and treatment strategy selection, great effort has been taken to develop assays for sensitive and accurate quantification of HER2. However, nonspecific amplification effect of most current assays limits the quantification accuracy of low abundance HER2. In the present work, we developed an LC-MS/MS-based quasi-targeted proteomics strategy coupled with hybridization chain reaction (HCR) for amplification of the HER2 protein signal. In the described strategy, the aptamer triggered the HCR system to undergo a cascade of hybridization events, with the two locked hairpins conjugated to the substrate peptide to form aptamer-HCR peptide probes. The membrane protein HER2 was recognized by probe and the signal was to be converted and then amplified into the mass response of the reporter peptide, which could be quantified using LC-MS/MS. The signal intensity was approximately five fold greater than that without signal amplification. Finally, the developed assay was applied for the quantitative analysis of HER2 in breast cell lines and monitor the dynamic change of HER2 in drug induced HER2 negative cells. The result demonstrated that combination of HCR signal amplification and mass spectrometry provides a novel approach for simple, accurate, and quantitative monitoring of low abundance protein.

HEADING AIMS:LncRNA HOXB-AS3 is proved as an oncogene in tumors. Herein, we determine the function and mechanism of HOXB-AS3 in epithelial ovarian cancer (EOC) cells.MATERIALS AND METHODS:Chi-square test, Kaplan-Meier (KM) analysis and Cox regression analysis were used to analyze the clinicopathological features of HOXB-AS3 in EOC patients. CCK8, transwell and wound healing assay were used to test the function of HOXB-AS3. Luciferase reporter assay, western blot and glycolysis rate assay were used for further mechanistic studies.KEY FINDINGS:HOXB-AS3 was abundantly expressed in EOC tissues, and higher levels of HOXB-AS3 in EOC patients were significantly associated with disease status and overall survival status. EOC patients with high levels of HOXB-AS3 had strikingly shorter disease-free survival (DFS) and overall survival (OS) times than those with low levels. HOXB-AS3 also might as an independent prognostic factor. Further study revealed knockdown of HOXB-AS3 significantly inhibited the proliferation, invasion and migration of EOC cells. Mechanistic investigations suggested that knockdown of HOXB-AS3 could decrease lactate dehydrogenase A (LDHA) expression and the extracellular acidification rate (ECAR) by sponging miR-378a-3p.SIGNIFICANCE:To our knowledge, this is the first study to suggest that HOXB-AS3 could crosstalk with miRNA in the cytoplasm and alter glycolysis in cancer cells. Our results improve our understanding of the mechanism of HOXB-AS3 and suggest that HOXB-AS3 can act as a predictor of OS and a target for EOC therapies.