scholarly journals SETDB1 promotes glioblastoma growth via CSF-1 - dependent macrophage recruitment by activating the AKT/mTOR signaling pathway

2020 ◽  
Author(s):  
Shuai Han ◽  
Wei Zhen ◽  
Tongqi Guo ◽  
Jianjun Zou ◽  
Fuyong Li
Keyword(s):  
Cell Proliferation ◽  
Tumor Microenvironment ◽  
Real Time ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Migration And Invasion ◽  
Common Disease ◽  
High Morbidity ◽  
Macrophage Recruitment ◽  
Syngeneic Mouse Model

Abstract Background: Glioblastoma is a common disease of the central nervous system (CNS), with high morbidity and mortality. In the infiltrate in the tumor microenvironment, tumor-associated macrophages (TAMs) are abundant, which are important factors in glioblastoma progression. However, the exact details of TAMs in glioblastoma progression have yet to be determined. Methods: The clinical relevance of SET domain bifurcated 1 (SETDB1) was analyzed by immunohistochemistry, real-time PCR and Western blotting of glioblastoma tissues. SETDB1-induced cell proliferation, migration and invasion were investigated by CCK-8 assay, colony formation assay, wound healing and Transwell assay. The relationship between SETDB1 and colony stimulating factor 1 (CSF-1), as well as TAMs recruitment was examined by Western blotting, real-time PCR and syngeneic mouse model.Results: Our findings showed that SETDB1 upregulated in glioblastoma and relative to poor progression. Gain and loss of function approaches showed the SETDB1 overexpression promotes cell proliferation, migration and invasion in glioblastoma cells. However, knockdown SETDB1 exerted opposite effects in vitro. Moreover, SETDB1 promotes AKT/mTOR-dependent CSF-1 induction and secretion, which leads to macrophage recruitment in the tumor, resulted in tumor growth. Conclusion: Our research clarified that SETDB1 regulates of tumor microenvironment and hence presents a potential therapeutic target for treating glioblastoma.

scholarly journals SETDB1 promotes glioblastoma growth via CSF-1-dependent macrophage recruitment by activating the AKT/mTOR signaling pathway

2020 ◽  
Vol 39 (1) ◽  
Author(s):  
Shuai Han ◽  
Wei Zhen ◽  
Tongqi Guo ◽  
Jianjun Zou ◽  
Fuyong Li
Keyword(s):  
Cell Proliferation ◽  
Tumor Microenvironment ◽  
Real Time ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Migration And Invasion ◽  
Common Disease ◽  
High Morbidity ◽  
Macrophage Recruitment ◽  
Syngeneic Mouse Model

Abstract Background Glioblastoma is a common disease of the central nervous system (CNS), with high morbidity and mortality. In the infiltrate in the tumor microenvironment, tumor-associated macrophages (TAMs) are abundant, which are important factors in glioblastoma progression. However, the exact details of TAMs in glioblastoma progression have yet to be determined. Methods The clinical relevance of SET domain bifurcated 1 (SETDB1) was analyzed by immunohistochemistry, real-time PCR and Western blotting of glioblastoma tissues. SETDB1-induced cell proliferation, migration and invasion were investigated by CCK-8 assay, colony formation assay, wound healing and Transwell assay. The relationship between SETDB1 and colony stimulating factor 1 (CSF-1), as well as TAMs recruitment was examined by Western blotting, real-time PCR and syngeneic mouse model. Results Our findings showed that SETDB1 upregulated in glioblastoma and relative to poor progression. Gain and loss of function approaches showed the SETDB1 overexpression promotes cell proliferation, migration and invasion in glioblastoma cells. However, knockdown SETDB1 exerted opposite effects in vitro. Moreover, SETDB1 promotes AKT/mTOR-dependent CSF-1 induction and secretion, which leads to macrophage recruitment in the tumor, resulted in tumor growth. Conclusion Our research clarified that SETDB1 regulates of tumor microenvironment and hence presents a potential therapeutic target for treating glioblastoma.

scholarly journals SETDB1 Promotes the Growth of Glioma via CSF-1-dependent Macrophage Recruitment by Activating the AKT/mTOR Signaling Pathway

2020 ◽  
Author(s):  
Shuai Han ◽  
Wei Zhen ◽  
Tongqi Guo ◽  
Jianjun Zou ◽  
fuyong li
Keyword(s):  
Real Time ◽  
Signaling Pathway ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Mtor Signaling ◽  
Migration And Invasion ◽  
High Morbidity ◽  
Mtor Signaling Pathway ◽  
Macrophage Recruitment ◽  
Syngeneic Mouse Model

Abstract Background: Glioma is a common disease of the central nervous system (CNS), with high morbidity and mortality. Among the infiltrates in the tumor microenvironment, tumor-associated macrophages (TAMs) are abundant and they are significant factors in glioma progression. However, the exact details of disease progression have yet to be determined. Methods: The clinical relevance of SETDB1 was analyzed by immunohistochemistry, real-time PCR and Western blotting and of glioma cancer tissues. Tumor cell proliferation, migration and invasion were investigated by MTS assay, colony formation assay, xenograft, wound healing and Transwell assay. The relationship between SETDB1 and CSF-1, as well as TAMS was examined by Western blotting, real-time PCR and syngeneic mouse model.Results: This work shows the presence and upregulation of SETDB1 in gliomaand its relationship with disease prognosis. Gain and loss of function approaches showed the inhibition of apoptosis and the promotion of growth, migration and invasion of the disease with SETDB1 overexpression and converse effects with SETDB1 silencing in vitro. Mechanistically, SETDB1 promotes CSF-1 expression by activating the AKT/mTOR signaling pathway. This leads to macrophage recruitment in the tumor, leading to tumor growth. Conclusion: This studyclarifies the modulation of tumor functions by SETDB1 and hence presents a future avenue for treating glioma.

miR-486-3p Controls the Apoptosis of Endometrial Carcinoma Cells

2022 ◽  
Vol 12 (5) ◽  
pp. 1002-1007
Author(s):  
Donghua Wang ◽  
Xiaoli Liu ◽  
Lirong Cao ◽  
Shixiong Gong ◽  
Yi He ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Migration ◽  
Endometrial Carcinoma ◽  
Real Time ◽  
Spectrophotometric Method ◽  
Real Time Pcr ◽  
Carcinoma Cells ◽  
Migration And Invasion ◽  
Invasive Capacity ◽  
Ec Cells

Our study aimed to discuss the mechanism of miR-486-3p in controlling the apoptosis of endometrial carcinoma (EC) cells. EC cells were divided into NC group, miR-486-3p mimic and miR-486-3p inhibitor group followed by analysis of miR-486-3p level by Real-time PCR, cell proliferation by spectrophotometric method, apoptosis by FCM, cell migration and invasion by Transwell analysis. EC cells showed reduced miR-486-3p level. The EC malignant biological behaviors could be prompted through retraining miR-486-3p level with increased EC cell invasive capacity. DDR1 was a target of miR-486-3p. The variation of tumor activity could be regulated through controlling DDR1 expression. In conclusion, the apoptotic and invasive characteristic of EC cells are restrained after overexpression of miR-486-3p in EC cells through targeting DDR1, indicating that miR-486-3p could be considered to be one kind of brand-new target for the treatment of EC.

scholarly journals MicroRNA-1253 Suppresses Cell Proliferation Migration and Invasion of Osteosarcoma by Targeting MMP9

2021 ◽  
Vol 20 ◽  
pp. 153303382199527
Author(s):  
Jianwen Mo ◽  
Tiansheng Zheng ◽  
Li Lei ◽  
Peng Dai ◽  
Jun Liu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
Migration And Invasion ◽  
Quantitative Real Time Pcr ◽  
Osteoblast Cells ◽  
Downstream Target ◽  
Cell Functions

Purpose: MicroRNAs play an important role in osteosarcoma (OS) development and progress. Although miR-1253 was considered as a tumor-inhibitor in some cancers, it’s function in the OS is not clear. Methods: In our study, we examined the expression of miR-1253 in OS cells and osteoblast cells using quantitative real-time PCR. The proliferation of OS cells was measured by BrdU assay, and we performed transwell to detect migration and invasion of OS cells. Meanwhile, EMT proteins were tested by western blot. We used Bioinformatics to predict the target genes of miR-1253 and found out Matrix metalloproteinases9 (MMP9) was one of that. The direct combination between miR-1253 and MMP9 was verified by double luciferase reporting experiment. Quantitative real-time PCR and western blot were used to detect the expression of MMP9. Results: We found that the expression level of miR-1253 in OS cells was significantly lower than that in osteoblast cells. Overexpression of miR-1253 could significantly inhibit OS cell proliferation, migration, invasion and EMT. And then, MMP9 was predicted as a downstream target of miR-1253 by Bioinformatics analysis. Further experiments showed that miR-1253 could reduce the protein level of MMP9 by directly binding to the 3’-UTR of MMP9. Afterward, we performed a rescue experiment, in which both MMP9 and miR-1253 were overexpressed. Compared with the groups overexpressed miR-1253 alone, cell proliferation, migration and invasion in co-overexpression groups were improved. Conclusions: In summary, these results suggested that miR-1253 down-regulated in OS cells, and could suppress the proliferation, migration and invasion of OS cells. Its molecular regulatory mechanism was that inhibits the expression of the downstream target gene MMP9 by directly binding, thus affect OS cell functions. Therefore, miR-1253 has the potential to become a biomarker and therapeutic target for OS therapy.

Dioscorea nipponica Attenuates Migration and Invasion by Inhibition of Urokinase-Type Plasminogen Activator through Involving PI3K/Akt and Transcriptional Inhibition of NF-κB and SP-1 in Hepatocellular Carcinoma

2016 ◽  
Vol 44 (01) ◽  
pp. 177-195 ◽  
Author(s):  
Ming-Ju Hsieh ◽  
Chao-Bin Yeh ◽  
Hui-Ling Chiou ◽  
Ming-Chang Hsieh ◽  
Shun-Fa Yang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Real Time ◽  
Plasminogen Activator ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Migration And Invasion ◽  
Inhibitory Effects ◽  
Type Plasminogen Activator ◽  
Urokinase Type Plasminogen Activator ◽  
Casein Zymography

High mortality and morbidity rates for hepatocellular carcinoma (HCC) in Taiwan primarily result from uncontrolled tumor metastasis. In our previous studies, we have reported that Dioscorea nipponica Makino extract (DNE) has anti-metastasis effects on human oral cancer cells. However, the effect of DNE on hepatoma metastasis have not been thoroughly investigated and remains poorly understood. To determine the effects of DNE on the migration and invasion in HCC cells we used a wound healing model, Boyden chamber assays, gelatin/casein zymography and Western blotting. Transcriptional levels of matrix metalloproteinase-9 (MMP-9) and urokinase-type plasminogen activator (u-PA) were detected by real-time PCR and promoter assays. In this study, DNE treatment significantly inhibited the migration/invasion capacities of Huh7 cell lines. The results of gelatin/casein zymography and Western blotting revealed that the activities and protein levels of the MMP-9 and u-PA were inhibited by DNE. Tests of the mRNA levels, real-time PCR, and promoter assays evaluated the inhibitory effects of DNE on u-PA expression in human hepatoma cells. A chromatin immunoprecipitation (ChIP) assay showed not only that DNE inhibits u-PA expression, but also the inhibitory effects were associated with the down-regulation of the transcription factors of NF-[Formula: see text]B and SP-1 signaling pathways. Western blot analysis also showed that DNE inhibits PI3K and phosphorylation of Akt. In conclusion, these results show that u-PA expression may be a potent therapeutic target in the DNE-mediated suppression of HCC invasion/migration. DNE may have potential use as a chemo-preventive agent against liver cancer metastasis.

scholarly journals Reduced KISS1 expression acts as a predictor for aggressive features of cervical carcinoma and promotes migration and invasion in cervical carcinoma cells

2020 ◽  
Author(s):  
Zhuo Wang ◽  
Min Wang ◽  
Hao Yu ◽  
Hongyu Wang ◽  
Can Wang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Western Blotting ◽  
Cervical Carcinoma ◽  
Carcinoma Cells ◽  
Migration And Invasion ◽  
Metastasis Suppressor ◽  
High Morbidity ◽  
Cervical Epithelial Cells ◽  
Low Expression

Abstract Background Cervical carcinoma causes high morbidity and mortality in patients largely due to its invasion and metastasis. Kisspeptin (KISS1) had been found as metastasis suppressor in many malignancies. But the clinical significance and biological functions of KISS1 in cervical carcinoma was not elaborated. Methods The expression levels of KISS1 were detected by quantitative real time polymerase chain reaction (qRT-PCR), Methylation specific PCR (MSP) analysis and western-blotting in cervical carcinoma tissues and cells. A eukaryotic expression plasmid was transfected into cervical epithelial cells in order to initiate KISS1 overexpression, and the endogenous was silenced in cervical epithelial cells using short hairpin RNA (shRNA) sequences. Transfection efficiency was validated by western blotting assays. To explore the effects of KISS1 on the metastatic phenotype of cervical carcinoma, the cell proliferation was examined by CCK8 assay, and the cell migration and invasion were detected by cell scratch and transwell invasion assays, respectively. Results We investigated the relevance between KISS1 expression with its methylation and clinicopathological features of cervical carcinoma. Low expression of KISS1 predicts a poor prognosis and was associated with lymph node metastasis and TNM stage. And methylation of KISS1 promoter, which was linked with KISS1 inactivation, was also found in most clinical samples of aggressive cervical carcinoma. In vitro cervical carcinoma C33A cells, KISS1 overexpression significantly inhibited cell proliferation, migration and invasion in cells. While KISS1 knockdown promoted proliferation, migration and invasion in cervical epithelial cell line CRL-2614. Conclusion In summary, low-expression of KISS1 play a suppressive role for proliferation, migration and invasion in cervical carcinoma cells. Its expression and methylation are associated with clinical progression of cervical carcinoma. Hence, KISS1 could be a potential diagnostic and prognostic marker as well as a new target for the treatment of cervical carcinoma.

scholarly journals PLK1 downregulation by RO3280 prevents cell proliferation, migration, and invasion via the Wnt/β-catenin pathway in prostate cancer

2021 ◽  
Author(s):  
Yongxue Ding ◽  
Zhe Zhang ◽  
Min Ju
Keyword(s):  
Prostate Cancer ◽  
Cell Proliferation ◽  
Cancer Cells ◽  
Real Time ◽  
Cell Lines ◽  
Cancer Cell ◽  
Western Blotting ◽  
Cancer Cell Lines ◽  
Migration And Invasion ◽  
Prostate Cancer Cells

IntroductionHigher expression levels of serine/threonine-protein kinase 1 (PLK1) are significantly associated with tumorigenesis and poor clinical prognoses. Consequently, PLK1 is considered a latent target in cancer treatment. We aimed to determine the cytotoxic effects of RO3280 on prostate cancer cells.Material and methodsPLK1 expression was investigated using real-time PCR and western blotting in prostate cancer tissues and paired normal tissues. Real-time cell analysis, cell counting kit-8 assays, and 5-ethynyl-2¢-deoxyuridine cell proliferation assays were applied for the examination of cell proliferation ability. Wound healing assays and transwell assays were used to assess the migratory and invasive abilities of the prostate cancer cell lines with or without RO3280 treatment. Moreover, the target genes and pathways were detected by transcriptomics RNA sequencing in the cells cultured in RO3280 and through a series of bioinformatics analyses. Finally, the Wnt/β-catenin pathway was screened out and verified by western blotting.ResultsWe observed the mRNA and protein overexpression of PLK1 in the prostate cancer cells and tissues. The inhibition of PLK1 by RO3280 significantly reduced the migratory, invasive, and proliferative properties of the RO3280-treated cancer cell lines compared with their controls. RO3280 mediated the inactivation of the Wnt/β-catenin pathway, and reduced the rates of cell proliferation, migration, and invasion in prostate cancer cells.ConclusionsThis study’s findings are significant owing to the identification of the specific anticancer mechanism of RO3280, which may have therapeutic effects. This trial provides clarity on the feasibility of the use of RO3280 as a cancer therapeutic agent for prostate cancer.

RPS24c Isoform Facilitates Tumor Angiogenesis Via Promoting the Stability of MVIH in Colorectal Cancer

2020 ◽  
Vol 20 (5) ◽  
pp. 388-395 ◽  
Author(s):  
Yue Wang ◽  
Youjun Wu ◽  
Kun Xiao ◽  
Yingjie Zhao ◽  
Gang Lv ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Ribosomal Protein ◽  
Real Time ◽  
Tumor Angiogenesis ◽  
Real Time Pcr ◽  
Molecular Mechanisms ◽  
Migration And Invasion ◽  
Binding Interaction ◽  
Tubule Formation ◽  
The Stability

Background: Colorectal cancer (CRC) is the second leading cause of death worldwide, and distant metastasis is responsible for the poor prognosis in patients with advanced-stage CRC. RPS24 (ribosomal protein S24) as a ribosomal protein, multiple transcript variant encoding different isoforms have been found for this gene. Our previous studies have demonstrated that RPS24 is overexpressed in CRC. However, the mechanisms underlying the role of RPS24 in tumor development have not been fully defined. Methods: Expression of RPS24 isoforms and lncRNA MVIH in CRC tissues and cell lines were quantified by real-time PCR or western blotting assay. Endothelial tube formation assay was performed to determine the effect of RPS24 on tumor angiogenesis. The cell viability of HUVEC was determined by MTT assay, and the migration and invasion ability of HUVEC were detected by transwell assay. PGK1 secretion was tested with a specific ELISA kit. Results: Here, we found that RPS24c isoform was a major contributor to tumor angiogenesis, a vital process in tumor growth and metastasis. Real-time PCR revealed that RPS24c isoform was highly expressed in CRC tissues, while other isoforms are present in both normal and CRC tissues with no statistical difference. Moreover the change of RPS24 protein level is mainly due to the fluctuation of RPS24c. Furthermore, we observed that silencing RPS24c could decrease angiogenesis by inhibiting tubule formation, HUVEC cell proliferation and migration. Additionally, we investigated the molecular mechanisms and demonstrated that RPS24c mRNA interacted with lncRNA MVIH, the binding-interaction enhanced the stability of each other, thereby activated angiogenesis by inhibiting the secretion of PGK1. Conclusion: RPS24c facilitates tumor angiogenesis via the RPS24c/MVIH/PGK1 pathway in CRC. RPS24c inhibition may be a novel option for anti-vascular treatment in CRC.

scholarly journals Long non-coding RNA CCDC183-AS1 acts AS a miR-589-5p sponge to promote the progression of hepatocellular carcinoma through regulating SKP1 expression

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
He Zhu ◽  
Hongwei Zhang ◽  
Youliang Pei ◽  
Zhibin Liao ◽  
Furong Liu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Proliferation ◽  
Cell Lines ◽  
Human Cancer ◽  
Quantitative Polymerase Chain Reaction ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
High Morbidity ◽  
Regulatory Relationship

Abstract Background Hepatocellular carcinoma (HCC) is a common type of malignant human cancer with high morbidity and poor prognosis, causing numerous deaths per year worldwide. Growing evidence has been demonstrated that long non-coding RNAs (lncRNAs) are closely associated with hepatocarcinogenesis and metastasis. However, the roles, functions, and working mechanisms of most lncRNAs in HCC remain poorly defined. Methods Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of CCDC183-AS1 in HCC tissues and cell lines. Cell proliferation, migration and invasion ability were evaluated by CCK-8 and transwell assay, respectively. Animal experiments were used to explore the role of CCDC183-AS1 and miR-589-5p in vivo. Bioinformatic analysis, dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to confirm the regulatory relationship between CCDC183-AS1, miR-589-5p and SKP1. Results Significantly upregulated expression of CCDC183-AS1 was observed in both HCC tissues and cell lines. HCC patients with higher expression of CCDC183-AS1 had a poorer overall survival rate. Functionally, overexpression of CCDC183-AS1 markedly promoted HCC cell proliferation, migration and invasion in vitro and tumor growth and metastasis in vivo, whereas the downregulation of CCDC183-AS1 exerted opposite effects. MiR-589-5p inhibitor counteracted the proliferation, migration and invasion inhibitory effects induced by CCDC183-AS1 silencing. Mechanistically, CCDC183-AS1 acted as a ceRNA through sponging miR-589-5p to offset its inhibitory effect on the target gene SKP1, then promoted the tumorigenesis of HCC. Conclusions CCDC183-AS1 functions as an oncogene to promote HCC progression through the CCDC183-AS1/miR-589-5p/SKP1 axis. Our study provided a novel potential therapeutic target for HCC patients.

scholarly journals CDX2 and Reg IV expression and correlation in gastric cancer

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dandan Chai ◽  
Huifen Du ◽  
Kesheng Li ◽  
Xueliang Zhang ◽  
Xiaoqin Li ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cell Migration ◽  
Cancer Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Ectopic Expression ◽  
Rank Correlation ◽  
Migration And Invasion ◽  
Gastric Cancer Cells ◽  
Cell Migration And Invasion

Abstract Background Ectopic expression of CDX2 is associated with the development and progression of gastric cancer. Previous studies showed that CDX2 may be an upstream regulator of Reg IV expression in gastric cancer, and our previous report showed that Reg IV upregulated SOX9 expression and enhanced cell migration and invasion in gastric cancer cells. However, the regulatory roles of CDX2 have not been clarified in gastric cancer, and the correlation between CDX2 and Reg IV requires further study. Methods CDX2 and Reg IV were examined in gastric cancer specimens and paired adjacent tissues via real-time PCR and immunohistochemistry (IHC). The association between CDX2 and Reg IV was assessed using the χ2-test and Spearman’s rank correlation. To verify their relationship, knockdown and exogenous expression of CDX2 or Reg IV were performed in AGS and MKN-45 gastric cancer cells, and their expression was subsequently analyzed via a real-time PCR and western blotting. Wound-healing and Transwell assays were used to examine migration and invasion in AGS and MKN-45 cells following CDX2 silencing or overexpression. Results A positive correlation was observed between CDX2 and Reg IV expression at the mRNA and protein levels in gastric cancer tissues. CDX2 silencing significantly downregulated Reg IV expression, and CDX2 overexpression significantly upregulated Reg IV expression in AGS and MKN-45 cells. Neither Reg IV silencing nor overexpression had any effect on CDX2 protein expression in AGS or MKN-45 cells, even though both affected the expression of CDX2 mRNA. Functionally, CDX2 silencing significantly inhibited cell migration and invasion, and CDX2 overexpression significantly promoted cell migration and invasion in AGS and MKN-45 cells. Conclusions Our findings demonstrate that CDX2 expression was positively correlated with that of Reg IV in gastric cancer, and CDX2 promoted cell migration and invasion through upregulation of Reg IV expression in AGS and MKN-45 cells.

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