SPP1 targeted by miR-4262 in gastric cancer functions as an enhancer of cell growth and correlates with dismal prognosis

2020 ◽  
Author(s):  
Junqing Wang ◽  
Fengjie Hao ◽  
Yuchen Yang ◽  
Xiaochun Fei ◽  
Xunhua Chen
Keyword(s):  
Gastric Cancer ◽  
Transcriptional Regulation ◽  
Cell Growth ◽  
Cell Model ◽  
Luciferase Reporter ◽  
Clinicopathologic Features ◽  
Over Expression ◽  
Dismal Prognosis ◽  
New Biomarkers ◽  
Post Transcriptional Regulation

Abstract Background Advanced gastric cancer (GC) induces diamal prognosis and high mortality. Discovery of new biomarkers or differentially expressed genes (DEGs) is serving for early diagnosis, prevention and therapautic treatmen in GC. In this study, by combining with biostatistics analysis, we aimed to verify the aberrant high expression and enhancing effects of SPP1 on GC, and to explore the probable relative post-transcriptional regulation. Methods Three datasets (GSE13911, GSE19826 and GSE27342) from NCBI GEO database were explored. SPP1 was screened out and detected in 105 real GC patients through immunohistochemistry analysis and RT-qPCR assay. The patients’ clinicopathologic features were collected and analyzed. The expression of SPP1 was examinated in three GC cell lines (MKN-45, AGS and SNU-16) . MKN-45 cell model with SPP1 depletd was constrcted through shRNA transfection. CCK8 assay, cell cycle detection and apoptosis rate calculation were conducted to evaluate the ability of cell growth. MiR-4262 was filtered out as a potential up-streaming regulator of SPP1 mRNA through bioinformatic prediction, and the dual-luciferase reporter assay was used for validation. Rescue experiment was introduced to confirm the post-transcriptional regulation. Results Thirteen DEGs increased in GC were selected, among which SPP1 was screened out for its significant over-expression in GC. SPP1 expression profile was validated in both the 105 real GC patients’ samples and three GC cell liens. High SPP1 expression was found significantly associated with the patients’ clinicopathologic features related to unideal prognosis, including tumor size, lymph node metastasis, local invasion grade and TNM stage. Depletion of SPP1 remarkably suppressed the GC cell growth. Whilst, microRNA-4262 was validated directly binding to the 3’-UTR of SPP1 mRNA in GC cells, degenerating the expression of SPP1. Conclusions SPP1 probably functions as an oncogenic gene in GC, and provides us a new biomarker in GC hopeful to promote GC prevention, diagnose and therapeutic treatment.

scholarly journals SPP1 targeted by miR-4262 in gastric cancer functions as an enhancer of cell growth and correlates with dismal prognosis

2020 ◽  
Author(s):  
Junqing Wang ◽  
Fengjie Hao ◽  
Yuchen Yang ◽  
Xiaochun Fei ◽  
Xunhua Chen
Keyword(s):  
Gastric Cancer ◽  
Transcriptional Regulation ◽  
Cell Growth ◽  
Cell Model ◽  
Luciferase Reporter ◽  
Clinicopathologic Features ◽  
Over Expression ◽  
Dismal Prognosis ◽  
New Biomarkers ◽  
Post Transcriptional Regulation

Abstract Background Advanced gastric cancer (GC) induces diamal prognosis and high mortality. Discovery of new biomarkers or differentially expressed genes (DEGs) is serving for early diagnosis, prevention and therapautic treatmen in GC. In this study, by combining with biostatistics analysis, we aimed to verify the aberrant high expression and enhancing effects of SPP1 on GC, and to explore the probable relative post-transcriptional regulation. Methods Three datasets (GSE13911, GSE19826 and GSE27342) from NCBI GEO database were explored. SPP1 was screened out and detected in 105 real GC patients through immunohistochemistry analysis and RT-qPCR assay. The patients’ clinicopathologic features were collected and analyzed. The expression of SPP1 was examinated in three GC cell lines (MKN-45, AGS and SNU-16) . MKN-45 cell model with SPP1 depletd was constrcted through shRNA transfection. CCK8 assay, cell cycle detection and apoptosis rate calculation were conducted to evaluate the ability of cell growth. MiR-4262 was filtered out as a potential up-streaming regulator of SPP1 mRNA through bioinformatic prediction, and the dual-luciferase reporter assay was used for validation. Rescue experiment was introduced to confirm the post-transcriptional regulation. Results Thirteen DEGs increased in GC were selected, among which SPP1 was screened out for its significant over-expression in GC. SPP1 expression profile was validated in both the 105 real GC patients’ samples and three GC cell liens. High SPP1 expression was found significantly associated with the patients’ clinicopathologic features related to unideal prognosis, including tumor size, lymph node metastasis, local invasion grade and TNM stage. Depletion of SPP1 remarkably suppressed the GC cell growth. Whilst, microRNA-4262 was validated directly binding to the 3’-UTR of SPP1 mRNA in GC cells, degenerating the expression of SPP1. Conclusions SPP1 probably functions as an oncogenic gene in GC, and provides us a new biomarker in GC hopeful to promote GC prevention, diagnose and therapeutic treatment.

scholarly journals MicroRNA-19b-3p suppresses gastric cancer development by negatively regulating neuropilin-1

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Yingfeng Wei ◽  
Sheng Guo ◽  
Jianhua Tang ◽  
Jianjun Wen ◽  
Huifen Wang ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cell Growth ◽  
Western Blot ◽  
Bioinformatics Analysis ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Functional Assay ◽  
Mrna Levels ◽  
Neuropilin 1

Abstract Background Gastric cancer (GC) remains one of the most common digestive malignancies worldwide and ranked third causes of cancer-related death. Mounting evidence has revealed that miRNAs exert critical regulatory roles in GC development. Methods Immunohistochemistry (IHC) and western blot assay were performed to determine the protein expression levels of neuropilin-1 (NRP1) and mRNA levels were confirmed by quantitative RT-PCR (qRT-PCR) in GC tissues. Kaplan–Meier analysis was performed to evaluate the prognostic value of NRP1 in GC. Knockdown of NRP1 was conducted to analyse its function in vitro and vivo. Luciferase reporter assay, western blot and qRT-qPCR were employed to identify the miRNAs which directly targeted NRP1. Furthermore, Bioinformatics analysis and experimental verification were used to explore the potential molecular mechanism and signalling pathway. Results In the current study, we revealed that NRP1 was highly expressed in GC tumor tissues and was associated with poor prognosis in GC patients. NRP1 knockdown inhibited GC cell growth, migration and invasion in vitro, while suppressed GC xenograft tumor development in vivo. Bioinformatics analysis predicted that miR-19b-3p down-regulated NRP1 expression by targeting its 3′-UTR. Functional assay demonstrated that miR-19b-3p inhibited GC cell growth, migration and invasion via negatively regulating NRP1. Overexpression NRP1 partially reversed the regulatory effect of miR-19b-3p. Moreover, we showed that miR-19b-3p/NRP1 axis regulated the epithelial-to-mesenchymal transition and focal adhesion in GC, which might contribute the GC development and progression. Conclusions Taken together, our findings suggest a regulatory network of miR-19b-3p/NRP1 in GC development. The miR-19b-3p/NRP1 axis might be further explored as a potential diagnostic and therapeutic target in GC.

scholarly journals miR-483-3p, Mediated by KLF9, Functions as Tumor Suppressor in Testicular Seminoma via Targeting MMP9

2021 ◽  
Vol 10 ◽  
Author(s):  
Lei Zhang ◽  
Yashi Ruan ◽  
Zhiqiang Qin ◽  
Xian Gao ◽  
Kai Xu ◽  
...  
Keyword(s):  
Cell Growth ◽  
Tumor Suppressor ◽  
Testicular Germ Cell Tumor ◽  
Tumor Stage ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Testicular Germ Cell ◽  
Testicular Seminoma ◽  
Over Expression ◽  
Upstream Regulator

BackgroundSeminoma (SEM) is the most frequent testicular germ cell tumor with a high incidence in young men. The present study aims to explore the function and regulatory mechanism of miR-483-3p in SEM.MethodsRT-qPCR was performed to investigate miR-483-3p levels in SEM tissues. The effect of miR-483-3p on TCam-2 cells was assessed by CCK-8, colony formation, cell migration, and invasion assays. Luciferase reporter assays were performed to investigate the interaction between miR-483-3p and MMP9, and then the recovery experiments were performed. Moreover, the potential upstream regulator of miR-483-3p was predicted based on JASPAR database.ResultsmiR-483-3p was down-regulated in SEM tissues versus paracancerous normal tissues. The expression level of miR-483-3p was significantly associated with tumor stage by RT-qPCR. Functionally, miR-483-3p over-expression suppressed cell growth, migration, and invasion in SEM cell lines. Mechanically, miR-483-3p negatively regulated MMP9 by directly binding to its 3′-UTR. The over-expression of miR-483-3p could reverse the promoting role of MMP9 over-expression on the proliferation, migration, and invasion of TCam-2 cells. Moreover, KLF9 was identified as a potential upstream regulator of miR-483-3p and functions as a tumor suppressor.ConclusionsIn general, our study suggested that miR-483-3p could inhibit the cell growth, migration, and invasion of testicular SEM by targeting MMP9. Moreover, KLF9 is an upstream positive regulator of miR-483-3p and also functions as a tumor suppressor in SEM.

scholarly journals Post-transcriptional regulation of BRG1 by FIRΔexon2 in gastric cancer

Oncogenesis ◽  
2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Guzhanuer Ailiken ◽  
Kouichi Kitamura ◽  
Tyuji Hoshino ◽  
Mamoru Satoh ◽  
Nobuko Tanaka ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Transcriptional Regulation ◽  
Post Transcriptional Regulation

Abstract 3829: Endothelial Cell Response to Hypoxia is Modulated by Microrna-210 Direct Targeting of Ephrin-A3

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Pasquale Fasanaro ◽  
Yuri D’Alessandra ◽  
Valeria Di Stefano ◽  
Roberta Melchionna ◽  
Sveva Romani ◽  
...  
Keyword(s):  
Cell Migration ◽  
Endothelial Cell ◽  
Cell Growth ◽  
Cell Response ◽  
Luciferase Reporter ◽  
Protein Coding ◽  
Over Expression ◽  
Endothelial Cell Response ◽  
Induced Apoptosis ◽  
Necessary And Sufficient

MicroRNAs (miRNAs) are small non-protein-coding RNAs that negatively modulate gene expression. In this study, miRNAs role in endothelial cell response to hypoxia was investigated. It was found that the expression of miR-210 progressively increased upon exposure to hypoxia. Interestingly, HIF1-alpha was necessary and sufficient for miR-210 induction, while HIF2-alpha was not necessary. miR-210 over-expression in normoxic endothelial cells increased the formation of capillary-like structures on Matrigel and VEGF-induced cell migration. Conversely, miR-210 blockade via LNA-anti-miRNA transfection decreased the formation of capillary-like structures stimulated by hypoxia and inhibited cell migration in response to VEGF. miR-210 over-expression did not affect endothelial cell growth in both hypoxia and normoxia. However, anti-miR-210 transfection inhibited cell growth and induced apoptosis, both in normoxia and in hypoxia. We found that one relevant target of miR-210 in hypoxia was Ephrin-A3, since miR-210 was necessary and sufficient to down-regulate its expression. Moreover, luciferase reporter assays showed that Ephrin-A3 was a direct target of miR-210. Ephrin-A3 modulation by miR-210 had significant functional consequences: indeed, the expression of an Ephrin-A3 allele that is not targeted by miR-210, prevented miR-210-mediated stimulation of both tubulogenesis and chemotaxis. A combination of proteomic and transcriptomic approaches has been undertaken, identifying potential direct and indirect targets of miR-210 action. We conclude that miR-210 induction is a crucial element of endothelial cell response to hypoxia, affecting cell survival, migration and differentiation.

scholarly journals Mir-483-3p, Mediated by KLF9, Functions as Tumor Suppressor in Testicular Seminoma via Targeting MMP9

2020 ◽  
Author(s):  
Lei Zhang ◽  
Yashi Ruan ◽  
Xian Gao ◽  
Kai Xu ◽  
Xiaokai Shi ◽  
...  
Keyword(s):  
Cell Growth ◽  
Tumor Suppressor ◽  
Testicular Germ Cell Tumor ◽  
Tumor Stage ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Testicular Germ Cell ◽  
Testicular Seminoma ◽  
Over Expression ◽  
Upstream Regulator

Abstract Background: Seminoma (SEM) is the most frequent testicular germ cell tumor with a high incidence in young men. The present study aims to explore the function and regulatory mechanism of miR-483-3p in SEM.Methods: RT-qPCR was performed to investigate miR-483-3p levels in SEM tissues. The effect of miR-483-3p on TCam-2 cells was assessed by CCK-8, colony formation, cell migration and invasion assays. Luciferase reporter assays were performed to investigate the interaction between miR-483-3p and MMP9 and then the recovery experiments were performed. Moreover, the potential upstream regulator of miR-483-3p was predicted based on JASPAR database.Results: miR-483-3p was down‐regulated in SEM tissues versus paracancerous normal tissues. The expression level of miR-483-3p was significantly associated with tumor stage by RT-qPCR. Functionally, miR-483-3p over-expression suppressed cell growth, migration and invasion in SEM cell lines. Mechanically, miR-483-3p negatively regulated MMP9 by directly binding to its 3’-UTR. The over-expression of miR-483-3p could reverse the promoting role of MMP9 over-expression on the proliferation, migration and invasion of TCam-2 cells. Moreover, KLF9 was identified as a potential upstream regulator of miR-483-3p and functions as a tumor suppressor.Conclusions: In general, our study suggested that miR-483-3p could inhibit the cell growth, migration and invasion of testicular SEM by targeting MMP9. Moreover, KLF9 is an upstream positive regulator of miR-483-3p and also functions as a tumor suppressor in SEM.

scholarly journals Orb-dependent polyadenylation contributes to PLP expression and centrosome scaffold assembly

2021 ◽  
Author(s):  
Junnan Fang ◽  
Dorothy Lerit
Keyword(s):  
Transcriptional Regulation ◽  
Rna Binding ◽  
Genome Instability ◽  
Cytoplasmic Polyadenylation ◽  
Over Expression ◽  
Pericentriolar Material ◽  
And Function ◽  
Post Transcriptional Regulation ◽  
Mrna Polyadenylation

As the microtubule-organizing centers (MTOCs) of most cells, centrosomes engineer the bipolar mitotic spindle required for error-free mitosis. Drosophila Pericentrin (PCNT)-like protein (PLP) is a key centrosome component that directs formation of a pericentriolar material (PCM) scaffold required for PCM organization and MTOC function. Here, we investigate the post-transcriptional regulation of plp mRNA. We identify conserved binding sites for cytoplasmic polyadenylation element binding (CPEB) proteins within the plp 3′-untranslated region and examine the role of the CPEB ortholog, oo18 RNA-binding protein (Orb), in plp mRNA regulation. Our data show Orb biochemically interacts with plp mRNA and promotes PLP protein expression. Loss of orb, but not orb2, diminishes PLP levels in embryonic extracts. Consequently, PLP localization to centrosomes and function in PCM scaffolding is compromised in orb mutant embryos, resulting in genome instability and embryonic lethality. Moreover, we find PLP over-expression can restore centrosome scaffolding and rescue the cell division defects caused by orb depletion. Our data suggest Orb modulates PLP expression at the level of plp mRNA polyadenylation and showcases the post-transcriptional regulation of core, conserved centrosomal mRNAs as critical for centrosome function.

scholarly journals miR8181 is Involved in the Cell Growth and Development Regulation of Saccharina Japonica

2020 ◽  
Author(s):  
Xiaoqi Yang ◽  
Xiu liang Wang ◽  
Jian ting Yao ◽  
Wei Li ◽  
Delin Duan
Keyword(s):  
Transcriptional Regulation ◽  
Cell Growth ◽  
Blue Light ◽  
Cleavage Site ◽  
Growth And Development ◽  
Target Genes ◽  
Saccharina Japonica ◽  
Degradome Sequencing ◽  
Post Transcriptional Regulation ◽  
Development Regulation

Abstract Background: Aureochrome, a blue-light receptor found in photosynthetic stramenopiles, plays an important role in brown algal growth and development. Aureochrome preserves the reversed effector-sensor domain for blue light reception and acts as the candidate optogenetic tool for light induced post-transcriptional regulation, but the inner rapid regulation of aureochrome remains to be studied. MicroRNA (miRNAs) of plant can cleavage the specific base-pairing site of mRNA by RNA interference mechanism, and such post-transcriptional regulation of miRNAs to photoreceptor has received attention due to the flexible regulation pathway. However, the targeting relationship between aureochrome and miRNA is unclear.Results: In this study, the potential regulatory network between miRNAs and aureochrome were explored by transcriptome and sRNA sequencing in Saccharina japonica. Our results found that 18 miRNAs perfectly paired with aureochrome. Among the screened miRNAs, miR8181-x was negatively correlated with aureochrome5 with high credibility and exhibited tissue-specific expression in S. japonica. Degradome sequencing detected the exact cleavage site of miR8181-x on aureochrome5, confirming their targeting regulation relationship. Among the 54 target genes of miR8181-x, nine genes of ABC transporters, E3 ubiquitin-protein protein ligase, Hsp90, Mx1, PetC, EF2, GSA, HAD-superfamily hydrolase and SET2 that exhibited similar expression with aureochrome5 competed with the same binding site, thus constructing the competing endogenous RNA network. Functional analysis of miR8181-x target genes revealed that regulation of cell differentiation and development was enriched, indicating the potential role of miR8181-x in the regulation of growth and development.Conclusion: Our study found that miR8181-x negatively regulated the expression of aureochrome5. The exact cleavage site in aureochrome5 were verified by degradome sequencing, confirming the targeting relationships. Functional enrichment of miR8181-x target genes revealed that miR8181-x involved in the cell growth and development regulation of S. japonica.

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