scholarly journals Selective exosome exclusion of miR-375 by glioma cells promotes glioma progression by activating the CTGF-EGFR pathway

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Xiangdong Xu ◽  
Yang Liu ◽  
Yan Li ◽  
Huajian Chen ◽  
Yuxuan Zhang ◽  
...  
Keyword(s):  
Human Peripheral Blood ◽  
Biological Significance ◽  
Xenograft Model ◽  
Glioma Cells ◽  
Cell Types ◽  
Migration And Invasion ◽  
Functional Link ◽  
Qrt Pcr ◽  
Orthotopic Xenograft Model ◽  
Transmission Electron

Abstract Background Exosomes are membrane-bound extracellular vesicles of 40–150 nm in size, that are produced by many cell types, and play an important role in the maintenance of cellular homeostasis. Exosome secretion allows for the selective removal of harmful substances from cells. However, it remains unclear whether this process also takes place in glioma cells. Methods Herein, the role of the tumour-suppressor miR-375 was explored in human glioma cells. Immunoblotting and qRT-PCR experiments demonstrated a functional link between miR-375 and its target, connectivetissuegrowthfactor (CTGF), which led to the identification of the underlying molecular pathways. The exosomes secreted by glioma cells were extracted by ultracentrifugation and examined by transmission electron microscopy. Exosomal expression of miR-375 was then analysed by qRT-PCR; while the exosome secretion inhibitor, GW4869, was used to examine the biological significance of miR-375 release. Moreover, the dynamics of miR-375 release by glioma cells was investigated using fluorescently labelled exosomes. Finally, exosomal miR-375 release was examined in an orthotopic xenograft model in nude mice. Results MiR-375 expression was downregulated in gliomas. MiR-375 suppressed glioma proliferation, migration, and invasion by inhibiting the CTGF-epidermalgrowthfactorreceptor (EGFR) signalling pathway. MiR-375-containing exosomes were also identified in human peripheral blood samples from glioma patients, and their level correlated with disease progression status. Exosomal miR-375 secretion impacted the CTGF-EGFR pathway activity. Once secreted, exosomal miR-375 was not taken back up by glioma cells. Conclusions Exosomal miR-375 secretion allowed for sustained activation of the CTGF-EGFR oncogenic pathway, promoting the proliferation and invasion of glioma cells. These findings enhance our understanding of exosome biology and may inspire development of new glioma therapies.

scholarly journals Selective exosome exclusion of miR-375 by glioma cells promotes glioma progression by activating the CTGF-EGFR pathway

2020 ◽  
Author(s):  
Xiangdong Xu ◽  
Yang Liu ◽  
Yan Li ◽  
Huajian Chen ◽  
Yuxuan Zhang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Human Peripheral Blood ◽  
Biological Significance ◽  
Xenograft Model ◽  
Growth Factor Receptor ◽  
Glioma Cells ◽  
Tissue Growth ◽  
Migration And Invasion ◽  
Functional Link ◽  
Qrt Pcr

Abstract Background: Exosomes are membrane-bound extracellular vesicles of 40–150 nm in size, that are produced by many cell types, and play an important role in the maintenance of cellular homeostasis. Exosome secretion allows for the selective removal of harmful substances from cells. However, it remains unclear whether this process also takes place in glioma cells.Methods: Herein, the role of the tumour-suppressor miR-375 was explored in human glioma cells. Immunoblotting and qRT-PCR experiments demonstrated a functional link between miR-375 and its target, connective tissue growth factor (CTGF), which led to the identification of the underlying molecular pathways. The exosomes secreted by glioma cells were extracted by ultracentrifugation and examined by transmission electron microscopy. Exosomal expression of miR-375 was then analysed by qRT-PCR; while the exosome secretion inhibitor, GW4869, was used to examine the biological significance of miR-375 release. Moreover, the dynamics of miR-375 release by glioma cells was investigated using fluorescently labelled exosomes. Finally, exosomal miR-375 release was examined in an orthotopic xenograft model in nude mice.Results: MiR-375 expression was downregulated in gliomas. MiR-375 suppressed glioma proliferation, migration, and invasion by inhibiting the CTGF-epidermal growth factor receptor (EGFR) signalling pathway. MiR-375-containing exosomes were also identified in human peripheral blood samples from glioma patients, and their level correlated with disease progression status. Exosomal miR-375 secretion impacted the CTGF-EGFR pathway activity. Once secreted, exosomal miR-375 was not taken back up by glioma cells.Conclusions: Exosomal miR-375 secretion allowed for sustained activation of the CTGF-EGFR oncogenic pathway, promoting the proliferation and invasion of glioma cells. These findings enhance our understanding of exosome biology and may inspire development of new glioma therapies.

scholarly journals Exosomal excretion of miR-3 7 5 promotes glioma progression by activating the CTGF-EGF R  pathway

2020 ◽  
Author(s):  
Xiangdong Xu ◽  
Yang Liu ◽  
Yan Li ◽  
Huajian Chen ◽  
Yuxuan Zhang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Human Peripheral Blood ◽  
Biological Significance ◽  
Xenograft Model ◽  
Growth Factor Receptor ◽  
Glioma Cells ◽  
Tissue Growth ◽  
Migration And Invasion ◽  
Functional Link ◽  
Qrt Pcr

Abstract Background: Exosomes are membrane-bound extracellular vesicles of 40–150 nm produced by many cell types, and playing an important role in the maintenance of cellular homeostasis. Exosome secretion allows for the selective removal of harmful substances from cells. Whether this process also takes place in glioma cells is unknown. Methods: The role of the tumour suppressor miR-375 was explored in human glioma cell lines. Immunoblotting and qRT-PCR experiments demonstrated a functional link between miR-375 and its target, connective tissue growth factor (CTGF), and led to identify the involved molecular pathways. The exosomes secreted by glioma cells were extracted by ultracentrifugation and examined by transmission electron microscopy. The exosomal expression of miR-375 was analyzed by qRT-PCR. The exosome secretion inhibitor, GW4869, was used to study the biological significance of miR-375 release. Moreover, the dynamics of miR-375 release by glioma cells was investigated by using fluorescently labelled exosomes. Finally, exosomal miR-375 release was studied in an orthotopic xenograft model in nude mice. Results: MiR-375 expression was downregulated in gliomas. MiR-375 suppressed glioma proliferation, migration, and invasion by inhibiting the CTGF-epidermal growth factor receptor (EGFR) signalling pathway. MiR-375-containing exosomes were also found in human peripheral blood samples from glioma patients, and their level was correlated with the status of disease progression. Exosomal miR-375 secretion had an impact on the activity of the CTGF-EGFR pathway. Once secreted, exosomal miR-375 was not reuptaken by glioma cells. Conclusions: Exosomal miR-375 secretion allowed for sustained activation of the CTGF-EGFR oncogenic pathway, promoting the proliferation and invasion of glioma cells. These findings enhance our understanding of exosome biology, and may inspire the development of new therapies for glioma.

scholarly journals Ribosomal Protein S27-Like is overexpressed in glioma cells and inhibition of RPS27L expression suspends the tumorigenesis

2020 ◽  
Author(s):  
Zhongzheng Sun ◽  
Hao Xue ◽  
Yan Wei ◽  
Shaobo Wang ◽  
Jianye Xu ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Xenograft Model ◽  
Glioma Cells ◽  
Tumor Grade ◽  
Migration And Invasion ◽  
Normal Brain ◽  
Orthotopic Xenograft Model ◽  
High Expression Group

Abstract Background: Ribosomal Protein S27-Like is an evolutionarily conserved ribosomal protein and the role of RPS27L influencing the malignance of several cancers has been reported. However, its effects on glioma were still unknown. This investigation aims to characterize the clinical significance and the biological functions of RPS27L in gliomas.Methods: TCGA databases were explored to analyze the correlation between RPS27L expression and the clinical characteristics of glioma patients. Immunohistochemical staining was performed on glioma cases and normal brain tissues. The function of RPS27L in glioma was further explored using U87MG and A172 cell lines and a orthotopic xenograft model of nude mice.Results: Data obtained from TCGA database showed higher expression of RPS27L in glioma than normal, and the overall survival was lower in the high expression group. Immunohistochemistry showed the expression levels of RPS27L were increased with the tumor grade rising in gliomas. Functional assays showed knockdown of RPS27L inhibited proliferation, cell cycle transition, migration and invasion, while promoted apoptosis. Data of western blot indicated that knockdown of RPS27L increased the level of p21，Bax and Cleaved Caspase-3 while decreased the level of CDK4, cyclinD1, cyclinE1, Bcl-2 and MMP2, MMP9 in glioma cells. In vivo, the growth of orthotopic glioma xenografts was suppressed by expression of RPS27L shRNA, and the tumors with RPS27L shRNA showed less aggressiveness and reduced expression of Ki67, Bcl-2 and MMP2. Conclusions: RPS27L is overexpressed in glioma cells. Knockdown of RPS27L could inhibit the proliferation, migration and invasion while promote apoptosis of glioma cells in vitro and in vivo. RPS27L might be a potential prognostic biomarker and possible target for future therapy in glioma.

scholarly journals Ribosomal Protein S27-Like is overexpressed in glioma cells and inhibition of RPS27L expression suspends the tumorigenesis

2020 ◽  
Author(s):  
Zhongzheng Sun ◽  
Hao Xue ◽  
Yan Wei ◽  
Shaobo Wang ◽  
Jianye Xu ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Xenograft Model ◽  
Glioma Cells ◽  
Tumor Grade ◽  
Migration And Invasion ◽  
Normal Brain ◽  
Orthotopic Xenograft Model ◽  
High Expression Group

Abstract Background: Ribosomal Protein S27-Like is an evolutionarily conserved ribosomal protein and the role of RPS27L influencing the malignance of several cancers has been reported. However, its effects on glioma were still unknown. This investigation aims to characterize the clinical significance and the biological functions of RPS27L in gliomas.Methods: TCGA databases were explored to analyze the correlation between RPS27L expression and the clinical characteristics of glioma patients. Immunohistochemical staining was performed on glioma cases and normal brain tissues. The function of RPS27L in glioma was further explored using U87MG and A172 cell lines and a orthotopic xenograft model of nude mice.Results: Data obtained from TCGA database showed higher expression of RPS27L in glioma than normal, and the overall survival was lower in the high expression group. Immunohistochemistry showed the expression levels of RPS27L were increased with the tumor grade rising in gliomas. Functional assays showed knockdown of RPS27L inhibited proliferation, cell cycle transition, migration and invasion, while promoted apoptosis. Data of western blot indicated that knockdown of RPS27L increased the level of p21，Bax and Cleaved Caspase-3 while decreased the level of CDK4, cyclinD1, cyclinE1, Bcl-2 and MMP2, MMP9 in glioma cells. In vivo, the growth of orthotopic glioma xenografts was suppressed by expression of RPS27L shRNA, and the tumors with RPS27L shRNA showed less aggressiveness and reduced expression of Ki67, Bcl-2 and MMP2. Conclusions: RPS27L is overexpressed in glioma cells. Knockdown of RPS27L could inhibit the proliferation, migration and invasion while promote apoptosis of glioma cells in vitro and in vivo. RPS27L might be a potential prognostic biomarker and possible target for future therapy in glioma.

The effect of lncRNA HOTAIR on chemoresistance of ovarian cancer through regulation of HOXA7

2018 ◽  
Vol 399 (5) ◽  
pp. 485-497 ◽  
Author(s):  
Siwei Liu ◽  
Huajiang Lei ◽  
Fangyuan Luo ◽  
Yilin Li ◽  
Lan Xie
Keyword(s):  
Ovarian Cancer ◽  
Tumor Growth ◽  
Cancer Cells ◽  
Nude Mice ◽  
Tumor Development ◽  
Xenograft Model ◽  
Tumor Xenograft ◽  
Migration And Invasion ◽  
Ovarian Cancer Cells ◽  
Qrt Pcr

AbstractThis study aimed at investigating the biological functions of long non-coding RNAs (lncRNAs) hox transcript antisense intergenic RNA (HOTAIR) in resistant ovarian cancer cells, exploring the regulation effect of HOTAIR onHOXA7, and investigating their influence on the chemosensitivity of ovarian cancer cells. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for the verification of HOTAIR expression in resistant and sensitive groups. How HOTAIR downregulation affected cell proliferation, migration and invasion, and apoptosis were determined using the MTT assay and the colony formation assay, the Transwell assay and flow cytometry analysis, respectively. Immunohistochemistry was used to inspect the protein expression of HOXA7 in resistant and sensitive ovarian cancer tissues. The regulation relationship between HOTAIR andHOXA7was investigated by qRT-PCR and Western blot. The effect of HOTAIR andHOXA7on tumor growth was confirmed by the tumor xenograft model of nude mice. By knocking downHOXA7, HOTAIR downregulation restrained the ovarian cancer deterioration in functional experiments. Silencing of HOTAIR andHOXA7could effectively inhibit tumor growth and increase chemosensitivity of ovarian tumors in nude mice. Downregulation of HOTAIR negatively affected the survival and activity of resistant ovarian cancer cells, and suppressed the expression ofHOXA7. Silencing of HOTAIR andHOXA7could increase the chemosensitivity of ovarian cancer cells, thus suppressing tumor development.

miR-56a Mediates the Wnt/β-Catenin Pathway to Promote the Efficacy of Radiation on Glioma

2021 ◽  
Vol 11 (10) ◽  
pp. 1955-1960
Author(s):  
Min Chen ◽  
Heping Zhou ◽  
Jun Mao ◽  
Zhihong Li ◽  
Zhengjiang Zha
Keyword(s):  
Glioma Cell ◽  
Glioma Cells ◽  
Migration And Invasion ◽  
Biological Processes ◽  
Array Analysis ◽  
Mirna Array ◽  
Qrt Pcr ◽  
The Impact ◽  
Glioma Cell Migration

Clarification of the miR-56a-mediated effect of Wnt/β-catenin pathway in glioma cells on radiosensitization. miRNA arrays were used to analyze the differential expression of miRNAs in biopsies from glioma patients. qRT-PCR to detect the levels of miR-56a and Wnt/β-catenin expressed in glioma cells and tissues. Evaluation of the impact of miR-56a on cell growth, invasion, and migrationforming ability by MTT assay and colony formation experiments. To analyze the involvement of miR-56a-mediated Wnt/β-catenin pathway in glioma biological processes and to examine the impact of miR-56a in glioma cell radiosensitivity. After miRNA array analysis, we found that miR-56a expression was significantly increased, and further studies showed that ectopic miR-56a expression in glial cells was sensitive to radiotherapy. miR-56a induction of Wnt/β-catenin promotes the upregulation of Parp in glioma cells. miR-56a can promote glioma cell migration and invasion in vitro as an important potential target for glioma disease.

Cell Classification in Human Peripheral Blood Using the Amnis ImageStream® System.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3826-3826
Author(s):  
Philip Morrissey ◽  
Thaddeus George ◽  
David Basiji ◽  
Keith Frost ◽  
William Ortyn ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Fluorescence Intensity ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Biological Significance ◽  
Morphological Characteristics ◽  
Cell Types ◽  
Morphometric Parameters ◽  
Clinical Diagnostics ◽  
Wide Range

Abstract Amnis Corporation’s ImageStream® system combines the sample handling and quantitative power of flow cytometry with high-resolution brightfield, darkfield, and fluorescence cellular imagery. The system simultaneously generates up to six images of each cell in flow and can acquire data sets consisting of tens of thousands of cells in just a few minutes, while offering fluorescence sensitivity equal to or better than flow cytometry. The image data are analyzed using Amnis’ IDEAS® software, which automatically calculates over 200 morphometric and photometric features and associated statistics for each cell, identifying unique cell groups based not only on their fluorescence intensity signature but also on their morphological characteristics. The software offers the ability to view the imagery associated with any cell in a scatter plot, perform “virtual cell sorts” of user-specified sub-populations, and generate custom features of biological significance (e.g. N/C ratio). The ImageStream platform’s ability to quantitate morphologic and immunofluorescent differences between very large numbers of cells in suspension make it particularly well suited for hematology. In the present study, human peripheral blood mononuclear cells were stained with a fluorescent DNA binding dye to reveal nuclear morphology, as well as fluorescently labeled mAb to various CD markers. Five images of each cell were acquired: brightfield (transmitted light), darkfield (laser side scatter), and three fluorescent colors for nuclear imagery and quantitation of the CD marker abundance. The object of the study was to identify morphometric parameters in the brightfield, darkfield, and nuclear imagery that would prove useful in hematologic cell type classification. The mAb to CD antigens provided a positive control for use in the evaluation of the of the various morphometric parameters. Parameters with discriminating power included cellular size and texture, darkfield intensity and granularity, and nuclear fluorescence intensity, texture, and shape. Cell types that could be automatically discriminated using these parameters in lieu of immunofluorescent markers included neutrophils, eosinophils, monocytes, and lymphocytes (including putative activated lymphocytes). In addition to forming the basis for an advanced ImageStream hematology platform, it is envisioned that the automated morphometric classification of blood cells will act as the foundation for a wide range of image-based cellular assays performed in peripheral blood (e.g. NF-kB translocation, apoptosis, mAb compartmentalization), allowing the differential quantitation of assay results in various cell types for the purposes of basic research, drug discovery, and clinical diagnostics.

scholarly journals FAM87A as a Competing Endogenous RNA of miR-424-5p Suppresses Glioma Progression by Regulating PPM1H

2021 ◽  
Vol 2021 ◽  
pp. 1-18
Author(s):  
Hua Xu ◽  
Haiping Zhang ◽  
Lina Tan ◽  
Yang Yang ◽  
Haiyun Wang ◽  
...  
Keyword(s):  
Cell Function ◽  
Glioma Cells ◽  
Vital Role ◽  
Migration And Invasion ◽  
Competing Endogenous Rna ◽  
Qrt Pcr ◽  
Rate Effects ◽  
And Migration ◽  
Proliferation And Migration ◽  
Glioma Progression

Far less has been unveiled about the functions of lncRNAs on cancers yet. Here, we reported that lncRNA FAM87A, as a ceRNA of miR-424-5p, played a vital role in glioma development. qRT-PCR result indicated that FAM87A was abnormally downregulated in glioma tissue and cells. Survival analysis suggested that the FAM87A expression was negatively correlated with the survival rate. Effects of FAM87A on human glioma cell lines were also analyzed by MTT, Edu, and transwell assays. FAM87A hastened proliferation and migration of glioma cells. MiR-424-5p, predicted target of FAM87A, was fostered in glioma, which was examined by qRT-PCR. A negative correlation was indicated between FAM87A and miR-424-5p. Results of bioinformatics, dual luciferase, and RIP assays unveiled that FAM87A and miR-424-5p act upon each other. In addition, miR-424-5p targeted 3 ′ -UTR of PPM1H. Also, effects of miR-424-5p/FAM87A on glioma cells were identified via the cell function experiments. FAM87A suppressed PPM1H by binding to miR-424-5p competitively, thereby restraining cell proliferation, migration, and invasion. Collectively, these findings illuminated a new mechanism for glioma progression. Therefore, FAM87A may act as a feasible target for glioma treatment.

scholarly journals Super-Enhancer-Associated TMEM44-AS1 Aggravated Glioma Progression by Forming a Positive Feedback Loop With Myc

2021 ◽  
Author(s):  
Erbao Bian ◽  
Xueran Chen ◽  
Li Cheng ◽  
Meng Cheng ◽  
Zhigang Chen ◽  
...  
Keyword(s):  
Positive Feedback ◽  
Colony Formation ◽  
Feedback Loop ◽  
Xenograft Model ◽  
Glioma Cells ◽  
Migration And Invasion ◽  
Positive Feedback Loop ◽  
Mouse Xenograft ◽  
Super Enhancer ◽  
Mouse Xenograft Model

Abstract BackgroundLong non-coding RNAs (lncRNAs) have been considered as one type of gene expression regulator for cancer development, but it is not clear how these are regulated. This study aimed to identify a specific lncRNA that promotes the glioma progression.MethodsRNA sequencing (RNA-seq) and quantitative real-time PCR were performed to screen differentially expressed genes. CCK-8, transwell migration, invasion assays and a mouse xenograft model were performed to determine the functions of TMEM44-AS1. Co-IP, ChIP, Dual-luciferase reporter assays, RNA pulldown and RNA immunoprecipitation assays were performed to study the molecular mechanism of TMEM44-AS1 and the downstream target.ResultsWe identified a novel lncRNA TMEM44-AS1, which was aberrantly expressed in glioma tissues, and that increased TMEM44-AS1 expression was correlated with malignant progression and poor survival for patients with glioma. Expression of TMEM44-AS1 increased the proliferation, colony formation, migration, and invasion of glioma cells. Knockdown of TMEM44-AS1 in glioma cells reduced cell proliferation, colony formation, migration and invasion, and tumor growth in a nude mouse xenograft model. Mechanistically, TMEM44-AS1 is directly bound to the SerpinB3, and sequentially activated Myc signaling; Myc transcriptionally induced TMEM44-AS1 and directly bound to the promoter and super-enhancer of TMEM44-AS1, thus forming a positive feedback loop with TMEM44-AS. Further studies demonstrated that Myc interacts with MED1 regulates the super-enhancer of TMEM44-AS1. More importantly, a novel small-molecule Myc inhibitor, Myci975, alleviated TMEM44-AS1-promoted the growth of glioma cells. Finally, TMEM44-AS1 activated IL-6 signaling by recruiting EGR1 to the promoter of IL-6 in glioma cells. ConclusionsOur study implicates a crucial role of the TMEM44-AS1-Myc axis in glioma progression and provides a possible anti-glioma therapeutic agent.

scholarly journals Circular RNA TAF4B Promotes Bladder Cancer Progression by Sponging miR-1298-5p and Regulating TGFA Expression

2021 ◽  
Vol 11 ◽  
Author(s):  
Xiaoting Zhang ◽  
Xiaofeng Li ◽  
Xian Fu ◽  
Mengli Yu ◽  
Guicheng Qin ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Xenograft Model ◽  
Circular Rna ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Mesenchymal Transition ◽  
Qrt Pcr ◽  
Mouse Xenograft Model

BackgroundBladder cancer (Bca) is the most common malignant tumor of the urinary system. Circular RNAs (circRNAs) have been recognized as key regulators in tumorigenesis. However, the molecular mechanisms underlying circRNAs involved in the progression of BCa remain largely unknown.MethodsWe detected the expression level of circular RNA TAF4B (circTAF4B) by qRT-PCR assay. Cell proliferation was evaluated by CCK-8 and colony formation assays. Wound healing and Transwell assays were performed to measure cell migration and invasion capability. Moreover, we performed qRT-PCR and western blotting assays to determine the expression levels of epithelial-mesenchymal transition (EMT) markers. A nuclear/cytoplasmic fractionation assay was used to measure the subcellular location of circTAF4B. RNA pull-down and dual-luciferase reporter assays were used to detect the target microRNA of circTAF4B. A mouse xenograft model was produced to analyze the effect of circTAF4B on the tumorigenesis of BCa.ResultsIn this study, we identified a novel circular RNA, circTAF4B, that is significantly upregulated in BCa and correlated with poor prognosis. Downregulated circTAF4B abolished the growth, metastasis and EMT process in BCa cells. Mechanistically, we found that circTAF4B facilitated the expression of transforming growth factor A (TGFA) by sponging miR-1298-5p. Finally, circTAF4B enhanced the proliferation and EMT process of BCa cells in vivo.ConclusionIn summary, our study demonstrated that circTAF4B played a carcinogenic role in the growth, metastasis, and EMT process of BCa by regulating the miR-1298-5p/TGFA axis. Thus, circTAF4B may become a diagnostic and therapeutic target for BCa.

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