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1.
Gland Surg ; 11(5): 892-912, 2022 May.
Article in English | MEDLINE | ID: mdl-35694087

ABSTRACT

Background: Breast cancer (BRCA) is the leading cause of cancer mortality among women, and it is associated with many tumor suppressors and oncogenes. There is increasing evidence that transcription factors (TFs) play vital roles in human malignancies, but TFs-based biomarkers for BRCA prognosis were still rare and necessary. This study sought to develop and validate a prognostic model based on TFs for BRCA patients. Methods: Differentially expressed TFs were screened from 1,109 BRCA and 113 non-tumor samples downloaded from The Cancer Genome Atlas (TCGA). Univariate Cox regression analysis was used to identify TFs associated with overall survival (OS) of BRCA, and multivariate Cox regression analysis was performed to establish the optimal risk model. The predictive value of the TF model was established using TCGA database and validated using a Gene Expression Omnibus (GEO) data set (GSE20685). A gene set enrichment analysis was conducted to identify the enriched signaling pathways in high-risk and low-risk BRCA patients. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the TF target genes were also conducted separately. Results: A total of 394 differentially expressed TFs were screened. A 9-TF prognostic model, comprising PAX7, POU3F2, ZIC2, WT1, ALX4, FOXJ1, SPIB, LEF1 and NFE2, was constructed and validated. Compared to those in the low-risk group, patients in the high-risk group had worse clinical outcomes (P<0.001). The areas under the curve of the prognostic model for 5-year OS were 0.722 in the training cohort and 0.651 in the testing cohort. Additionally, the risk score was an independent prediction indicator for BRCA patients both in the training cohort (HR =1.757, P<0.001) and testing cohort (HR =1.401, P=0.001). It was associated with various cancer signaling pathways. Ultimately, 9 overlapping target genes were predicted by 3 prediction nomograms. The GO and KEGG enrichment analyses of these target genes suggested that the TFs in the model may regulate the activation of some classical tumor signaling pathways to control the progression of BRCA through these target genes. Conclusions: Our study developed and validated a novel prognostic TF model that can effectively predict 5-year OS for BRCA patients.

2.
Mol Biol Rep ; 49(4): 2711-2721, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35059968

ABSTRACT

OBJECTIVE: Breast cancer is one of the most common malignant and highly heterogeneous tumors in women. MicroRNAs (miRNAs), such as miR-1246, play important roles in various types of malignant cancers, including triple-negative breast cancer (TNBC). However, the biological role of miR-1246 in TNBC has not yet been fully elucidated. In this study, we studied the role of miR-1246 in the occurrence and development of TNBC and its mechanism of action. METHODS: Cell Counting Kit-8 (CCK-8), wound healing, and Transwell assays were performed to observe the effects of miR-1246 on TNBC cell proliferation, migration, and invasion, respectively. The expression of epithelial-mesenchymal transition (EMT) markers was detected by western blotting. Dual luciferase reporter assays were performed to determine whether DYRK1A is a novel target of miR-1246. In addition, an immunoprecipitation experiment was performed to verify the binding of DYRK1A to PGRN. Rescue experiments were performed to determine whether DYRK1A is a novel target of miR-1246 and whether miR-1246 suppresses the metastasis of breast cancer cells by targeting the DYRK1A/PGRN axis to prevent the epithelial-mesenchymal transition. RESULTS: Our results show that miR­1246 suppresses the proliferation, migration, and invasion of TNBC cells, DYRK1A is a novel target of miR-1246 and Importin-8 mediated miR-1246 nuclear translocation. MiR­1246 plays a suppressive role in the regulation of the EMT of TNBC cells by targeting DYRK1A. DYRK1A mediates the metastasis of triple-negative breast cancer via activation of the EMT. We identified PGRN as a novel DYRK1A-interacting protein. Overexpression of PGRN and DYRK1A promoted cell proliferation and migration of TNBC, but this effect was reversed by co-expression of miR-1246 mimics.DYRK1A and PGRN act together to regulate the occurrence and development of breast cancer through miR-1246. CONCLUSION: MiR-1246 suppresses the metastasis of breast cancer cells by targeting the DYRK1A/PGRN axis and preventing the epithelial-mesenchymal transition. The MiR-1246/DYRK1A/PGRN axis regulates TNBC progression, suggesting that MiR-1246 could be promising therapeutic targets for the treatment of TNBC.


Subject(s)
MicroRNAs , Progranulins , Protein Serine-Threonine Kinases , Protein-Tyrosine Kinases , Triple Negative Breast Neoplasms , Cell Line, Tumor , Cell Movement/genetics , Epithelial-Mesenchymal Transition , Female , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , Progranulins/metabolism , Protein Serine-Threonine Kinases/metabolism , Protein-Tyrosine Kinases/metabolism , Triple Negative Breast Neoplasms/genetics , Triple Negative Breast Neoplasms/metabolism , Triple Negative Breast Neoplasms/pathology , Dyrk Kinases
3.
Front Cell Dev Biol ; 9: 679185, 2021.
Article in English | MEDLINE | ID: mdl-34095151

ABSTRACT

Objective: In this study, we focused on the potential mechanism of miRNAs carried by human umbilical cord mesenchymal stem cells-derived exosomes (hUCMSCs-exo) in breast cancer (BC). Methods: RT-qPCR was conducted for the expression of miR-224-5p and HOXA5 in tissues and cells. After co-culture of exosomes and MCF-7 or MDA-MB-231 cells, the cell proliferation was observed by MTT and cell colony formation assay, while apoptosis was measured by flow cytometry. In addition, the expression of HOXA5 and autophagy pathway-related proteins LC3-II, Beclin-1 and P62 was detected by western blotting. And immunofluorescence was applied for detection of LC3 spots. The binding of miR-224-5p to HOXA5 was verified by the luciferase reporter gene assay and RNA-binding protein immunoprecipitation assay. Finally, in vivo experiment was performed to investigate the effect of miR-224-5p on BC growth. Results: MiR-224-5p was up-regulated and HOXA5 was down-regulated in BC tissues and cells. HOXA5 was confirmed to be the target gene of miR-224-5p. MiR-224-5p carried by hUCMSCs-exo was able to promote the proliferation and autophagy of BC cells, while inhibited apoptosis. Bases on xenograft models in nude mice, it was also revealed that miR-224-5p carried by hUCMSCs-exo could regulate autophagy and contribute to the occurrence and development of BC in vivo. Conclusion: MiR-224-5p carried by hUCMSCs-exo can regulate autophagy via inhibition of HOXA5, thus affecting the proliferation and apoptosis of BC cells.

4.
Onco Targets Ther ; 14: 2433-2448, 2021.
Article in English | MEDLINE | ID: mdl-33859479

ABSTRACT

INTRODUCTION: Breast cancer is the main reason for cancer-related deaths in women and the most common malignant cancer among women. In recent years, immunosuppressive factors have become a new type of treatment for cancer. However, there are no effective biomarkers for breast cancer immunotherapy. Therefore, exploring immune-related biomarkers is presently an important topic in breast cancer. METHODS: Gene expression profile data of breast cancer from The Cancer Genome Atlas (TCGA) was downloaded. Scale-free gene co-expression networks were built with weighted gene co-expression network analysis. The correlation of genes was performed with Pearson's correlation values. The potential associations between clinical features and gene sets were studied, and the hub genes were screened out. Gene Ontology and gene set enrichment analysis were used to reveal the function of hub gene in breast cancer. The gene expression profiles of GSE15852, downloaded from the Gene Expression Omnibus database, were used for hub gene verification. In addition, candidate biomarkers expression in breast cancer was studied. Survival analysis was performed using Log rank test and Kaplan-Meier. Immunohistochemistry was used to analyze the expression of CCNA2. RESULTS: A total of 6 modules related to immune cell infiltration were identified via the average linkage hierarchical clustering. According to the threshold criteria (module membership >0.9 and gene significance >0.35), a significant module consisting of 13 genes associated with immune cells infiltration were identified as candidate hub genes after performed with the human protein interaction network. And 3 genes with high correlation to clinical traits were identified as hub genes, which were negatively associated with the overall survival. Among them, the expression of CCNA2 was increased in metastatic breast cancer compare with non-metastatic breast cancer, who underwent immunotherapy. Immunohistochemistry results showed that CCNA2 expression in carcinoma tissues was elevated compared with normal control. DISCUSSION: CCNA2 identified as a potential immune therapy marker in breast cancer, which were first reported here and deserved further research.

5.
Clin Lab ; 67(4)2021 Apr 01.
Article in English | MEDLINE | ID: mdl-33865261

ABSTRACT

BACKGROUND: Mycoplasma hominis (MH) is an opportunistic pathogen, which often causes funisitis, spontaneous abortion, and low birth weight. However, current laboratory methods are time-consuming, labor-intensive, or require specialized laboratory instruments. Recombinase polymerase amplification (RPA) technology is a rapidly developing field because of the significance for clinical application and commercial value. Few studies have reported the use of RPA to detect MH. In this study, we developed the rapid MH detection assay, which may be potentially used as a sensitive point-of-care testing (POCT) in clinic. METHODS: Primers based on the MH 16SrRNA gene and gap gene were explored and screened out. The probe of RPA-LFD was designed based on the optimal primer and confirmed. The reaction conditions of temperature and time for RPA were optimized. The sensitivity and specificity of the analysis were explored. A total of 60 clinical specimens were used to verify the efficiency of the two methods. RESULTS: The optimal reaction conditions were determined as 15 minutes and 39°C. The sensitivity of RPA was 10-6 ng for MH, which is 100,000 times more sensitive than traditional PCR. Moreover, we observed another six non-target reproductive tract common pathogens without amplification products. Furthermore, we found that there was no significant difference between RPA and the cultivation method (p > 0.05). These two methods were in good agreement (κ = 0.938) when detecting clinical specimens. CONCLUSIONS: A new method for sensitive and rapid detection of MH based on RPA was successfully developed, which can be applied in large-scale screening and as a supplementary method to classical methods.


Subject(s)
Mycoplasma hominis , Recombinases , Humans , Mycoplasma hominis/genetics , Nucleic Acid Amplification Techniques , Polymerase Chain Reaction , Recombinases/genetics , Sensitivity and Specificity
6.
Cell Death Dis ; 12(1): 94, 2021 01 18.
Article in English | MEDLINE | ID: mdl-33462208

ABSTRACT

Circular RNAs (circRNAs) is one type of important non-coding RNAs that participate in tumorigenesis and cancer progression. In our previous study, we performed a microarray analysis of circRNAs between the tumor tissues and the adjacent normal tissues of hepatocellular carcinoma (HCC) patients, and found that the circRNA hsa_circ_0007456 is significantly downregulated in the tumor tissues and correlated with the prognosis of HCC. We further investigated the relationship between the expression levels of hsa_circ_0007456 in HCC and the susceptibility of NK cells, and found that the expression levels of hsa_circ_0007456 in HCC cell lines significantly influenced their susceptibility to NK cells. Through a series of screening and validation, we found that hsa_circ_0007456 mainly functioned through sponging miR-6852-3p and regulating the expression of intercellular adhesion molecule-1 (ICAM-1) in HCC. The miR-6852-3p/ICAM-1 axis is essential for the NK cytotoxicity toward HCC mediated by hsa_circ_0007456. In conclusion, we identify here hsa_circ_0007456 as a promising biomarker of HCC, and highlight hsa_circ_0007456/miR-6852-3p/ICAM-1 axis as an important signaling pathway in the process of tumor immune evasion and the tumorigenesis of HCC.


Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Hepatocellular/genetics , Intercellular Adhesion Molecule-1/metabolism , Killer Cells, Natural/metabolism , Liver Neoplasms/genetics , Carcinoma, Hepatocellular/pathology , Cell Proliferation , Female , Humans , Liver Neoplasms/pathology , Male , Middle Aged , RNA, Circular , Transfection
7.
Ann Clin Lab Sci ; 50(4): 519-527, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32826250

ABSTRACT

OBJECTIVE: Triple-negative breast cancer (TNBC) is one of the most common malignant, highly heterogeneous tumors in women. MicroRNAs (miRNAs), such as miR-200c, play an important role in various types of malignant cancer, including TNBC. However, the biological role of miRNA-200c in TNBC is not well understood. In this study, we investigated the mechanism of miR-200c in the growth of TNBC. METHODS: Reverse transcription quantitative polymerase chain reaction was used to detect the expression of miR-200c in TNBC tissues and TNBC cells. Cell Counting Kit-8 (CCK-8) assays, wound healing, and transwell assays were used to observe the effects of miR-200c on TNBC cell proliferation, migration, and invasion, respectively. The expression of epithelial-mesenchymal transition (EMT) markers were detected by Western blotting. Dual luciferase reporter assays were used to test whether ZEB2 is a novel target of miR-200c. RESULT: Our results show that ZEB2 is a novel target of miR-200c and that ZEB2 mediates the metastasis of triple-negative breast cancer via EMT. CONCLUSION: miR-200c attenuates TNBC cell invasion and EMT by targeting ZEB2. Our data therefore suggest that miR-200c may be used to develop novel early-stage diagnostic and therapeutic strategies for TNBC.


Subject(s)
MicroRNAs/genetics , Triple Negative Breast Neoplasms/genetics , Zinc Finger E-box Binding Homeobox 2/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Epithelial-Mesenchymal Transition/genetics , Epithelial-Mesenchymal Transition/physiology , Female , Gene Expression Regulation, Neoplastic/genetics , Homeodomain Proteins/genetics , Humans , MicroRNAs/metabolism , Neoplasm Metastasis/genetics , Repressor Proteins/genetics , Triple Negative Breast Neoplasms/metabolism , Zinc Finger E-box Binding Homeobox 2/genetics
8.
J Neurosci ; 40(13): 2644-2662, 2020 03 25.
Article in English | MEDLINE | ID: mdl-32066583

ABSTRACT

Yes-associated protein (YAP) transcriptional coactivator is negatively regulated by the Hippo pathway and functions in controlling the size of multiple organs, such as liver during development. However, it is not clear whether YAP signaling participates in the process of the formation of glia scars after spinal cord injury (SCI). In this study, we found that YAP was upregulated and activated in astrocytes of C57BL/6 male mice after SCI in a Hippo pathway-dependent manner. Conditional knockout (KO) of yap in astrocytes significantly inhibited astrocytic proliferation, impaired the formation of glial scars, inhibited the axonal regeneration, and impaired the behavioral recovery of C57BL/6 male mice after SCI. Mechanistically, the bFGF was upregulated after SCI and induced the activation of YAP through RhoA pathways, thereby promoting the formation of glial scars. Additionally, YAP promoted bFGF-induced proliferation by negatively controlling nuclear distribution of p27Kip1 mediated by CRM1. Finally, bFGF or XMU-MP-1 (an inhibitor of Hippo kinase MST1/2 to activate YAP) injection indeed activated YAP signaling and promoted the formation of glial scars and the functional recovery of mice after SCI. These findings suggest that YAP promotes the formation of glial scars and neural regeneration of mice after SCI, and that the bFGF-RhoA-YAP-p27Kip1 pathway positively regulates astrocytic proliferation after SCI.SIGNIFICANCE STATEMENT Glial scars play critical roles in neuronal regeneration of CNS injury diseases, such as spinal cord injury (SCI). Here, we provide evidence for the function of Yes-associated protein (YAP) in the formation of glial scars after SCI through regulation of astrocyte proliferation. As a downstream of bFGF (which is upregulated after SCI), YAP promotes the proliferation of astrocytes through negatively controlling nuclear distribution of p27Kip1 mediated by CRM1. Activation of YAP by bFGF or XMU-MP-1 injection promotes the formation of glial scar and the functional recovery of mice after SCI. These results suggest that the bFGF-RhoA-YAP-p27Kip1 axis for the formation of glial scars may be a potential therapeutic strategy for SCI patients.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Astrocytes/metabolism , Cell Cycle Proteins/metabolism , Gliosis/metabolism , Nerve Regeneration/physiology , Spinal Cord Injuries/metabolism , Adaptor Proteins, Signal Transducing/genetics , Animals , Cell Cycle Proteins/genetics , Cell Proliferation/physiology , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Gliosis/genetics , Gliosis/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Neurons/metabolism , Recovery of Function/physiology , Signal Transduction/physiology , Spinal Cord Injuries/genetics , Spinal Cord Injuries/pathology , YAP-Signaling Proteins
9.
Breast Cancer ; 27(4): 573-585, 2020 Jul.
Article in English | MEDLINE | ID: mdl-31960363

ABSTRACT

BACKGROUND: Long non-coding RNA (lncRNA) T-cell leukemia/lymphoma 6 (TCL6) has been reported as a potential tumor suppressor. However, its expression and function in breast cancer remain unknown. This study was performed to investigate the expression of lncRNA TCL6 in breast cancer and its clinical significance. METHODS: The survival and clinical molecular roles of TCL6 in breast cancer were analyzed. The underlying mechanism modulated by TCL6 and its correlation with immune-infiltrating cells were investigated. Gene Expression Omnibus (GEO) datasets were further used to confirm the prognostic role of TCL6. RESULTS: TCL6 low expression was not correlated with age, clinical stage, T stage, lymph node metastasis, distant metastasis, human epidermal growth factor 2 status, but was associated with estrogen receptor and progesterone receptor (PR) status and was an independent factor for worse survival (HR 1.876, P = 0.016). Specifically, low TCL6 expression correlated with worse prognosis in PR-negative patients. TCL6 could predict worse survival in luminal B breast cancer based on intrinsic subtypes. Immune-related pathways such as Janus kinase-signal transducer of activators of transcription were regulated by TCL6. Further finding revealed that TCL6 correlated with immune infiltrating cells such as B cells (r = 0.25, P < 0.001), CD8+ T cells (r = 0.23, P < 0.001), CD4+ T cells (r = 0.25, P < 0.001), neutrophils (r = 0.21, P < 0.001), and dendritic cells (r = 0.27, P < 0.001). TCL6 was also positively correlated with tumor-infiltrating lymphocytes infiltration and PD-1, PD-L1, PD-L2, and CTLA-4 immune checkpoint molecules (P < 0.001). CONCLUSION: Our findings suggest that lncRNA TCL6 correlates with immune infiltration and may act as a useful prognostic molecular marker in breast cancer.


Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/genetics , Gene Expression Regulation, Neoplastic/immunology , Lymphocytes, Tumor-Infiltrating/immunology , RNA, Long Noncoding/metabolism , Adult , Aged , Aged, 80 and over , Breast/immunology , Breast/pathology , Breast Neoplasms/immunology , Breast Neoplasms/mortality , Breast Neoplasms/pathology , Datasets as Topic , Female , Gene Expression Profiling , Humans , Kaplan-Meier Estimate , Middle Aged , Oligonucleotide Array Sequence Analysis
10.
DNA Cell Biol ; 38(12): 1529-1539, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31647329

ABSTRACT

Because of the phenotypic and molecular diversity, it is still difficult to predict breast cancer prognosis. This study aimed to develop and validate a multi-lncRNA (long noncoding RNA) signature to improve the survival prediction for breast cancer. Three hundred twenty-seven breast cancer patients from GSE20685 were used as a training set. GSE88770 including 117 patients and The Cancer Genome Atlas datasets including 1077 patients were used as 2 validation sets. Kaplan-Meier curve, the LASSO (least absolute shrinkage and selection operator) method, univariate and multivariate Cox analyses were applied to build a molecular model for predicting survival. Function analysis of this lncRNA signature was investigated. A novel eight-lncRNA molecular signature was first identified from multiple datasets. This signature classified patients into the high-risk and low-risk groups. Breast cancer in the high-risk group showed significantly worse survival than that in the low-risk group. Further multivariate Cox analysis revealed that this molecular signature was an independent prognostic factor for breast cancer in the training and validation sets. Furthermore, stratification analyses showed that this molecular signature was also used to classify patients into the low- and high-risk groups in patients with low or high T-stage, patients with or without lymph node metastasis, older or younger, estrogen receptor-positive or -negative, and progesterone receptor-positive or -negative patients. Our eight-lncRNA signature was a powerful tool in predicting prognosis in Luminal B breast cancer based on molecular subtype. This lncRNA signature involved in cell adhesion, apoptosis, cell differentiation, and immune regulation. Our study provided a reliable eight-lncRNA molecular signature for survival prediction of breast cancer, and this signature can stratify patients into the high- and low-risk groups. This molecular signature may help the selection of the suitable treatment strategies.


Subject(s)
Biomarkers, Tumor/genetics , Breast Neoplasms/mortality , Nomograms , RNA, Long Noncoding/genetics , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Female , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Lymphatic Metastasis , Middle Aged , Prognosis , Risk Factors , Survival Rate
11.
Anal Biochem ; 577: 14-20, 2019 07 15.
Article in English | MEDLINE | ID: mdl-30991018

ABSTRACT

BACKGROUND: Insulin is central to regulating fat and carbohydrate metabolism in the body. However, it is difficult to detect insulin using mass spectrometry (MS). The integration of nanotechnology with mass spectrometry for selective and sensitive detection is an important research area. Our aim was to establish a method to detect insulin using silicon nanoparticle-assisted high-throughput MS. METHODS: Different nanomaterials with the potential for use as MALDI components to enhance the MS signal by increasing peptide ionization were investigated in the present study. Insulin in samples was enriched with antibody-coated silicon nanoparticles and then analyzed by MALDI-TOF MS. Method validation was performed in the present study. RESULTS: A platform for insulin detection with small sample volumes (100 µL) and a simplified procedure was successfully developed. The silicon nanoparticle-MS assay exhibited high sensitivity (LOQ, 0.1 nM) and good linear correlation of MS intensity with insulin concentration (R2 = 0.99). Intra-assay precision (% coefficient of variation) ranged from 1.81 to 4.53%, and interassay precision ranged from 2.71 to 8.09%. In addition, a correlation between the MALDI assay and a chemiluminescence immunoassay (CIA) was completed in patient samples, and the resulting Deming regression revealed good agreement (R2 = 0.981). CONCLUSIONS: In our study, we found that the insulin signal could be enhanced with silicon nanoparticles. A new insulin determination method, immunoaffinity-based mass spectrometry, that saves time and involves simple processes, has been successfully established. The present assay was validated to detect insulin with low limits of detection.


Subject(s)
Insulin/blood , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Humans , Nanoparticles/chemistry , Silicon/chemistry
12.
Biochem Biophys Res Commun ; 506(4): 793-798, 2018 12 02.
Article in English | MEDLINE | ID: mdl-30389135

ABSTRACT

BACKGROUND/AIMS: Autophagy is known as a protective intracellular procedure, which can be regulated by several factors. MiRNA has been suggested as a potential element to mediate autophagy pathway in carcinomas. Our study was aim to investigate the role of autophagy in breast cancer cells and identify the involved molecular mechanism METHODS: The expression of LC3I/II, SQSTM1 and Smad4 were detected by western blot. The mRNA level were quantified by real-time PCR. MDC staining was used to directly visualize autophagosome formation. Target Scan 7.2 was used to predict biological targets of miR-224-5p RESULTS: MiR-224 -5p expression was upregulated in metastatic breast cancer and non-metastatic breast cancer cells compare with control. Moreover, miR-224-5p inhibition enhanced cellular autophagy levels in breast cancer cells. MiR-224-5p could suppress Smad4 expression in MDA-MB-231 cells, which indicated that Smad4 was identified as a target of miR-224-5p in breast cancer cells with high metastatic potential CONCLUSIONS: Our study revealed that miR-224-5p inhibited autophagy by targeting Smad4 in MDA-MB-231 cells. The results indicated that miR-224-5p/Smad4 regulating autophagy might be a novel regulatory network contributing to metastasis of breast cancer. MiR-224-5p and Smad4 is involved in breast tumorigenesis, which is possibly a novel target for breast cancer therapy.


Subject(s)
Autophagy , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , MicroRNAs/metabolism , Smad4 Protein/metabolism , Adult , Base Sequence , Breast Neoplasms/blood , Breast Neoplasms/genetics , Case-Control Studies , Cell Line, Tumor , Female , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Neoplasm Metastasis
13.
Int J Oncol ; 53(2): 567-578, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29901191

ABSTRACT

Colorectal cancer (CRC) is a type of cancer with a mortality rate among the highest worldwide owing to its high rate of metastasis. Therefore, inflammation-associated metastasis in the development of CRC is currently a topic of considerable interest. In the present study, the pro-inflammatory cytokine interleukin-4 (IL-4) was identified to promote the epithelial-mesenchymal transition (EMT) of CRC cells. However, the enhancing effect of IL-4 was more evident in HCT116 cells compared with in RKO cells. Accordingly, an increased expression level of STAT6 was observed in HCT116 cells compared with RKO cells. Further investigations identified that E2F1 was required for maintaining the level of signal transducer and activator of transcription 6 (STAT6) in HCT116 cells. Mechanistically, E2F1 induced specificity protein 3 (SP3) directly by binding to the promoter of the STAT6 gene and activating its transcription in CRC cells. As a result, phosphorylation-activated STAT6 increased the expression of several EMT drivers, including zinc finger E-box-binding homeobox (Zeb)1 and Zeb2, which serve a critical function in IL-4-induced EMT. Rescue experiments further confirmed that IL-4-induced EMT relied on an intact E2F1/SP3/STAT6 axis in CRC cells. Finally, analysis of clinical CRC specimens revealed a positive correlation between E2F1, SP3 and STAT6. The ectopically expressed E2F1/SP3/STAT6 axis indicated a poor prognosis in patients with CRC. In conclusion, the E2F1/SP3/STAT6 pathway was identified to be essential for IL-4 signaling-induced EMT and aggressiveness of CRC cells.


Subject(s)
Colorectal Neoplasms/metabolism , E2F1 Transcription Factor/metabolism , Interleukin-4/metabolism , STAT6 Transcription Factor/genetics , Sp3 Transcription Factor/metabolism , Animals , Cell Proliferation , Colorectal Neoplasms/genetics , Epithelial-Mesenchymal Transition , Female , Gene Expression Regulation, Neoplastic , HCT116 Cells , HT29 Cells , Humans , Male , Mice , Neoplasm Transplantation , Promoter Regions, Genetic , STAT6 Transcription Factor/metabolism , Signal Transduction
14.
Cell Mol Neurobiol ; 38(5): 1067-1079, 2018 Jul.
Article in English | MEDLINE | ID: mdl-29441488

ABSTRACT

Interneurons in the olfactory bulb (OB) are generated from neuronal precursor cells migrating from anterior subventricular zone (SVZa) not only in the developing embryo but also throughout the postnatal life of mammals. In the present study, we established an in vivo electroporation assay to label SVZa cells of rat both at embryonic and postnatal ages, and traced SVZa progenitors and followed their migration pathway and differentiation. We found that labeled cells displayed high motility. Interestingly, the postnatal cells migrated faster than the embryonic cells after applying this assay at different ages of brain development. Furthermore, based on brain slice culture and time-lapse imaging, we analyzed the detail migratory properties of these labeled precursor neurons. Finally, tissue transplantation experiments revealed that cells already migrated in subependymal zone of OB were transplanted back into rostral migratory stream (RMS), and these cells could still migrate out tangentially along RMS to OB. Taken together, these findings provide an in vivo labeling assay to follow and trace migrating cells in the RMS, their maturation and integration into OB neuron network, and unrecognized phenomena that postnatal SVZa progenitor cells with higher motility than embryonic cells, and their migration was affected by extrinsic environments.


Subject(s)
Brain/cytology , Brain/embryology , Cell Movement , Electroporation/methods , Animals , Animals, Newborn , Cell Differentiation , Cell Shape , Green Fluorescent Proteins/metabolism , Lateral Ventricles/cytology , Neural Stem Cells/cytology , Olfactory Bulb/cytology , Rats, Sprague-Dawley , Staining and Labeling
15.
Biochem Biophys Res Commun ; 493(2): 1018-1025, 2017 11 18.
Article in English | MEDLINE | ID: mdl-28942143

ABSTRACT

ARID1A as a subunit of SWI/SNF chromatin complexes is frequently mutated in human pancreatic cancer, however its exact role in pancreatic tumorigenesis remain unclear. In this study, we investigated the effects of ARID1A loss on human pancreatic epithelial cell lines HPNE, BxPC-3 with KRAS mutant (KRASG12D) expression. We found that ARID1A knockdown promoted cell proliferation and colony formation in cooperation with active mutant KRASG12D. Function assay revealed that ARID1A knockdown accelerated cell cycle progression, and repressed KRASG12D-induced cell senescence. Transcriptome analysis revealed ARID1A knockdown led to miR-503 upregulation. CDKN2A was identified as a target of miR-503, which contributes to cell senescence. Thus, our data suggests that ARID1A deficiency promote KRASG12D-driven pancreatic tumorigenesis through miR-503/CDKN2A-mediated senescence.


Subject(s)
Cell Transformation, Neoplastic/pathology , Cyclin-Dependent Kinase Inhibitor p18/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Nuclear Proteins/metabolism , Pancreas/pathology , Pancreatic Neoplasms/pathology , Transcription Factors/metabolism , Carcinogenesis/genetics , Carcinogenesis/metabolism , Carcinogenesis/pathology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/metabolism , Cellular Senescence , Cyclin-Dependent Kinase Inhibitor p16 , Cyclin-Dependent Kinase Inhibitor p18/metabolism , DNA-Binding Proteins , Gene Expression Profiling , Humans , MicroRNAs/metabolism , Nuclear Proteins/genetics , Pancreas/metabolism , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/metabolism , Point Mutation , Proto-Oncogene Proteins p21(ras)/genetics , Proto-Oncogene Proteins p21(ras)/metabolism , RNA Interference , RNA, Small Interfering/genetics , Transcription Factors/genetics
16.
Oncol Lett ; 12(1): 269-274, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27347136

ABSTRACT

The purpose of the present study was to investigate the serum levels of microRNA (miRNA/miR)-382-3p, -598-3p, -1246 and -184 in breast cancer patients and to assess their feasibility as biomarkers for breast cancer screening. Serum samples were obtained from 100 breast cancer patients and 40 age-matched healthy control subjects in Taizhou Central Hospital (Taizhou, Zhejiang, China) between January 2013 and September 2014. The serum expression levels of miR-382-3p, -598-3p, -1246 and -184 were determined by stem-loop reverse transcription-quantitative polymerase chain reaction. Receiver operating characteristic curves were drawn to evaluate the sensitivity and specificity of the serum miRNA expression levels for the screening of breast cancer. miR-382-3p and -1246 were significantly upregulated in the serum of the breast cancer patients, while miR-598-3p and -184 were significantly downregulated. The sensitivity and specificity to detect breast cancer were as follows: miR-382-3p, 52.0 and 92.5%; miR-598-3p, 95.0 and 85.0%; miR-1246, 93.0 and 75.0%; and miR-184, 87.5 and 71.0%, respectively. The expression levels of the four serum miRNAs were not correlated with the patients' clinical stage. In summary, miR-382-3p, -598-3p, -1246 and -184 are all involved in the development of breast cancer, and are promising biomarkers for breast cancer detection.

17.
J Drug Target ; 24(3): 224-32, 2016.
Article in English | MEDLINE | ID: mdl-26219518

ABSTRACT

BACKGROUND: Peptide-mediated drug delivery system (DDS) has been increasingly used to promote on-demand treatment efficacy of cancers. Herein, LTLRWVGLMS (LS10) peptide is selected as the functional ligand for specific glioma-targeting drug delivery. LS10 peptide selectively binds to NG2 proteins that are widely overexpressed in the glioma cells and restricted in normal tissue. LS10 peptide-decorated DDS is expected to hold vast promises in glioma therapy and decrease unwanted side effects. METHODS: LS10 peptide was conjugated on the surface of poly(ethylene glycol)-poly(ɛ-caprolactone) (PEG-PCL) nanoparticles via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide coupling reaction. Using U87MG cells as the glioma cell model, cellular uptake, internalization mechanism, cellular cytotoxicity and apoptosis were investigated. 1,1'-Dioctadecyl-3,3,3',3'-tetramethyl indotricarbocyanine iodide were used as fluorescence probes to investigated in vivo glioma targeting capability of LS10-NP. The glioma therapeutic efficacy of paclitaxel-loaded LS10-NP was studied on glioma-bearing nude mice. RESULTS: The LS10-NP with size of 119 nm enhanced cellular uptake on U87MG cells, increased cytotoxicity of the loaded paclitaxel (PTX), and improved penetration in 3D U87MG glioma spheres. In vivo biodistribution experiments showed that LS10-NP exhibited the enhanced drug localization at glioma site, which resulted in prolonged survival time of glioma-bearing mice. CONCLUSION: Our results indicated that LS10 peptide-modified nanoparticulate DDS could significantly improve the anti-glioma efficacy.


Subject(s)
Drug Delivery Systems/methods , Ethylene Glycols/chemistry , Glioma/drug therapy , Nanoparticles/administration & dosage , Nanoparticles/chemistry , Peptides/administration & dosage , Peptides/chemistry , Polyesters/chemistry , Animals , Apoptosis/drug effects , Cell Line, Tumor , Ethylene Glycols/administration & dosage , Ethylene Glycols/pharmacokinetics , Glioma/metabolism , Male , Mice , Mice, Nude , Paclitaxel/administration & dosage , Paclitaxel/therapeutic use , Particle Size , Peptides/pharmacokinetics , Polyesters/administration & dosage , Polyesters/pharmacokinetics , Tissue Distribution
18.
Zhong Yao Cai ; 39(6): 1300-2, 2016 Jun.
Article in Chinese | MEDLINE | ID: mdl-30156802

ABSTRACT

Objective: To study the chemical constituents of Swertia binchuanensis. Methods: The constituents were separated and purifyed by conventional methods. Their structures were identified on the basis of spectral analysis such as 1H-NMR,13C-NMR. Results: Seven compounds were isolated and identified as 1, 2, 3, 4-tetrahydro-1, 4, 8-trihydroxy-6-methoxyxanhone( Ⅰ), erythrocentaurin( Ⅱ),1-O-ß-D-glucopyranosyl-1, 2, 3, 4-tetrahydro-4, 8-dihydroxy-6-methoxyxanthone( Ⅲ), maslinic acid( Ⅳ),1-hydroxy-3, 7-dimethoxyxanthone( Ⅴ),1, 8-dihydroxy-3, 5-dimethoxyxanthone( Ⅵ),1, 7-dihydroxy-3, 4, 8-termethoxyxanthone( Ⅶ). Conclusion: All compounds were isolated from Swertia binchuanensis for the first time.


Subject(s)
Swertia , Drugs, Chinese Herbal , Flavonoids , Molecular Structure , Triterpenes
19.
Int J Clin Exp Med ; 8(8): 12956-62, 2015.
Article in English | MEDLINE | ID: mdl-26550214

ABSTRACT

OBJECTIVE: To study the expression and molecular mechanism of long noncoding RNAs (lncRNA) including HEIH and HULC in proliferation and invasion of hepatoma cells. METHODS: We detected the expression of HEIH and HULC in hepatocellular carcinoma cell line (MHCC97L and HepG2), as well as in human normal hepatocyte line (cHL-7702) by real-time PCR. Using MTT and transwell, we investigated the effect of HEIH and HULC on proliferation and invasion of hepatoma cells with siRNA and expression plasmid. To explore the molecular mechanism, we use western blot to reveal the role of HEIH and HULC in tumor invasion related gene expression. RESULTS: The expression of HEIH and HULC in hepatocellular carcinoma cell line was significantly increased compared with human normal hepatocyte line (P<0.05). The expression of HULC in HepG2 was higher than that in MHCC97L. The over-expression of HULC could enhance proliferation of MHCC97L and HepG2, however, the over-expression of HEIH could not. The over-expression of HULC and HEIH could promote invasion of MHCC97L and HepG2. Invasion of MHCC97L and HepG2 did not have significant change after down-regulating of HEIH and HULC by siRNA. Over-expression of HULC up-regulated the expression of Snail in HepG2. CONCLUSIONS: The expression of HEIH and HULC increased significantly in hepatocellular carcinoma cell line compared with that in human normal hepatocyte line. HULC could promote proliferation of hepatoma cells. HEIH and HULC play an important role in the invasion of hepatocellular carcinoma cell.

20.
Zhonghua Yi Xue Za Zhi ; 94(28): 2216-21, 2014 Jul 22.
Article in Chinese | MEDLINE | ID: mdl-25331476

ABSTRACT

OBJECTIVE: To explore the effect of extracts of Prunella vulgaris L.on proteome of human lung adenocarcinoma cell line A549 by two-dimensional electrophoresis and mass spectrometry and elucidate the mechanism of anti-lung adenocarcinom effect of Prunella vulgaris L.at the level of proteome. METHODS: The proliferative activity of human lung adenocarcinoma cell line A549 was evaluated by methyl thiazolyl tetrazolium (MTT) colorimetric assay. According to the difference of culture medium, all subjects were divided into the experimental group with culture medium of extracts of Prunella vulgaris L. (300 µg/ml) and the control group with culture medium of DMSO (0.3%). Proteins were isolated by two-dimensional electrophoresis and proteomic maps acquired by silver staining. And proteomic analysis was processed by Image Master 2D Quant Platinum 6.0. The proteins with > 2-fold differences were used to analyze by mass spectrometry and confirmed by Western blot. RESULTS: The expressions of inositol 1, 4, 5-triphosphate receptor-interacting protein-like 2 precursor, heat shock cognate protein 70, serine-threonine kinase receptor-associated protein, tropomyosin 2(ß) isoform 1, cyclin B3, MED12L protein and macrophin 1 isoform 2 were higher in experimental group than those in control group (ratio (medicial/normal) 2.051 93, 1 000 001, 2.203 08, 5.042 01, 15.178 00, 1 000 001, 1 000 001) . And the expressions of enolase 1, M2-type pyruvate kinase, heat shock protein 27, Rho GDP-dissociation inhibitor 1, heat shock protein ß1, TapasinERP57 heterodimer chain A, inorganic pyrophosphatase and mitochondrial Cysteinyl-tRNA synthetase 2 (putative) were lower in experimental group than those in control group (ratio (medicial/normal) 0.485 18, 0.491 53, 0.465 43, 0.454 71, 0.499 34, 0.450 36, 0.494 62, 0.437 33). CONCLUSIONS: The extracts of Prunella vulgaris L.have multi-target and multi-pathway effects on anti-lung adenocarcinoma. And its possible mechanisms may be due to the regulation of steady state of calcium ion, cell cycle and its steady state and the inhibition of tumor cell proliferation and metastasis.


Subject(s)
Adenocarcinoma/pathology , Lung Neoplasms/pathology , Plant Extracts/pharmacology , Prunella/chemistry , Adenocarcinoma of Lung , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Humans , Protein Serine-Threonine Kinases , Proteome , Proteomics
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