Interferon Regulatory Factor 2 (IRF2) Inhibits the Invasion and Migration of Renal Clear Cell Carcinoma Cells by Downregulation of Spindle Pole Body Component 24 (SPC24)

2021 ◽  
Vol 11 (10) ◽  
pp. 1881-1890
Author(s):  
Bing Lu ◽  
Hongbo Xu ◽  
Meng Ding ◽  
Chunyin Yan
Keyword(s):  
Clear Cell ◽  
Mrna Level ◽  
Spindle Pole Body ◽  
Kidney Tissue ◽  
Spindle Pole ◽  
Adenocarcinoma Cell Line ◽  
Invasion And Migration ◽  
Body Component ◽  
And Migration ◽  
Migration Of Cells

It has been reported that the increased expression of SPC24 (spindle pole body component 24) was involved in the initiation and development of various cancers. However, the role of SPC24 in ccRCC (clear cell renal cell carcinoma) remains largely unknown. In the present study, the changes and correlation of SPC24 and IRF2 (interferon regulatory factor 2) with ccRCC were evaluated by using GEPIA, TCGA and GTEx database. Then the involvement of SPC24 and IRF2 in invasion and migration was investigated in CaKi-1 cells, a human renal adenocarcinoma cell line. The bioinformatics assay revealed that the expression of SPC24 and IRF2 in kidney tissue of patients with renal clear cell cancer was significantly increased, and the expression of SPC24 and IRF2 in kidney tissue was positively and negatively related to cancer phase and survival rate in patients with ccRCC respectively. Notably, in vitro experimental study demonstrated that SPC25 promoted the invasion and migration of CaKi-1 cells, a human renal adenocarcinoma cell line. Furthermore, IRF2 shows potential binding site with SPC24 promoter, IRF2 overexpression significantly decreased SPC24 mRNA level, whereas inhibition of IRF2 with specific small hairpin RNA (shRNA) significantly increased SPC24 mRNA level. Functionally, inhibition of SPC24 with specific shRNA reversed the stimulatory effect of IRF2 shRNA on the invasion and migration of cells, whereas SPC24 over-expression reversed the inhibitory effect of IRF2 overexpression on the invasion and migration of cells. Finally, ChIP (chromatin immunoprecipitation) assay shows that IRF2 could directly bind with SPC24 promoter. In conclusion, these results demonstrated that IRF2/SPC24 signaling pathway contributes to the increased invasion and migration in ccRCC.

scholarly journals RNA-Binding Protein IGF2BP1 Enhances mRNA Stability and Translation Efficiency of INHBA to Promote the Invasion and Migration of Esophageal Squamous Cancer Cells

2021 ◽  
Author(s):  
Juan-Juan Wang ◽  
Ding-Xiong Chen ◽  
Yu Zhang ◽  
Xin Xu ◽  
Yan Cai ◽  
...  
Keyword(s):  
Mrna Stability ◽  
Rna Binding ◽  
Binding Protein ◽  
Mrna Level ◽  
Rna Stability ◽  
Translation Efficiency ◽  
Invasion And Migration ◽  
Squamous Cancer ◽  
And Migration

Abstract BackgroundMetastasis are mainly responsible for the death of patients with advanced esophageal squamous cell carcinoma (ESCC). At present, there is no targeted drug for the treatment of ESCC in clinic practice. The present study aims to investigate the roles and implication of IGF2BP1 overexpression in ESCC.MethodsIGF2BP1 protein expression was assessed by immunohistochemistry (IHC), and the mRNA abundance of IGF2BP1 and INHBA were analyzed with TCGA datasets and by RNA in situ hybridization (RISH). Cell viability, migration, invasion and in vivo metastasis assays were performed to explore the roles of IGF2BP1 in ESCC. RNA immunoprecipitation sequencing (RIP-seq) and mass spectrometry were applied to identify the targets and interacting proteins of IGF2BP1, respectively. RIP-PCR, RNA-pulldown, immunofluorescence (IF), gene specific m6A PCR and RNA stability assay were used to uncover the molecular mechanism of IGF2BP1 dysregulation. The methylation level of IGF2BP1 promoter region was detected by MSP-PCR. BTYNB, a small molecular inhibitor which could block the binding of IGF2BP1 to c-Myc mRNA, was evaluated for the inhibition effect on the malignant phenotypes of ESCC cells.ResultsIGF2BP1 overexpression was detected in ESCC tissues and associated with depth of tumor invasion. Knockdown of IGF2BP1 inhibited ESCC cell invasion and migration as well as tumor metastasis. Importantly, INHBA was identified as a direct target of IGF2BP1 in ESCC cells, which had a role in promoting the malignant phenotypes. TCGA data and RISH analyses showed that the mRNA level of INHBA was upregaluted in ESCC tissues as well. Mechanistically, IGF2BP1 bound and stabilized INHBA mRNA and then enhanced its translation, leading to an activation of Smad2/3 signaling. Ras GTPase-activating protein-binding protein 1 (G3BP1) was recruited by IGF2BP1 to participate in activating the signaling process, which was inhibited by the IGF2BP1 inhibitor BTYNB. Of note, IGF2BP1 mRNA expression in ESCC cells was negatively correlated with the level of its promoter methylation.ConclusionsIGF2BP1 overexpression promotes the invasion and migration of ESCC cells by up-regulating TGF-β-Smad2/3 pathway through enhancing INHBA mRNA stability and translation, providing a potential therapeutic target for ESCC treatment.

scholarly journals Overexpression of Spondin-2 Is Associated with Recurrence-Free Survival in Patients with Localized Clear Cell Renal Cell Carcinoma

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Hui-Min Ma ◽  
Meng Yu ◽  
Cong Wu ◽  
Hou-Bao Huang ◽  
Ya-Wei Li ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Clear Cell ◽  
Recurrence Free Survival ◽  
Cell Renal Cell Carcinoma ◽  
Free Survival ◽  
Invasion And Migration ◽  
And Migration

Background. The spondin-2 (SPON2) gene is overexpressed in multiple malignant tumors and may promote tumor aggressiveness. However, its expression profile and functional roles in clear cell renal cell carcinoma (ccRCC) are still unclear. Methods. SPON2 expression in ccRCC was evaluated using expression data from TCGA and GEO databases, then confirmed by local patient population (94 patients). The clinical significance of SPON2 expression was evaluated. Downregulation of SPON2 was performed using small-interfering RNA (siRNA). The effects of SPON2 silencing on cell proliferation, apoptosis, invasion, and migration in vitro were investigated. Results. SPON2 was overexpressed in the majority of the ccRCC at both mRNA and protein levels. SPON2 expression was significantly correlated with stage, grade, and recurrence (all P<0.05) in patients with localized ccRCC. The receiver operating characteristic (ROC) curve showed that SPON2 expression could serve as a predictor of recurrence. SPON2 expression was significantly associated with recurrence-free survival (RFS) in patients with localized ccRCC. Knocking down SPON2 resulted in suppressed cell invasion and migration in vitro. Conclusion. SPON2 expression might function as a prognostic biomarker in patients with localized ccRCC.

Effect of EH domain containing protein 2 on the biological behavior of clear cell renal cell carcinoma

2019 ◽  
Vol 38 (8) ◽  
pp. 927-937 ◽  
Author(s):  
C Liu ◽  
S Liu ◽  
L Wang ◽  
Y Wang ◽  
Y Li ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Western Blot ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Clear Cell ◽  
Control Group ◽  
Cell Renal Cell Carcinoma ◽  
Invasion And Migration ◽  
Eh Domain ◽  
And Migration

To investigate the effects of EH domain containing protein 2 (EHD2) on clear cell renal cell carcinoma (ccRCC) and provide new insights for the clinical treatment of rental cancer. Forty patients (26 males and 14 females, 62.4 ± 5.7 years old) with ccRCC were selected from January 2015 to December 2016 to serve as research subjects in this study. The EHD2 protein expression in the tumor tissues and adjacent healthy tissues of ccRCC patients were detected by Western Blot assay. The cells of ccRCC cell lines RLC-310 and 786-O were divided into normal control group (control), no-load control group (pLV), EHD2 overexpression group (pLV-EHD2), and EHD2 interference group (pLV-siEHD2). The expression levels of EHD2 protein in each group of cells were detected by western blot. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. Wound healing assay was performed to check the cell migration ability. Transwell invasion assay was used to detect the cell invasion ability. Cell apoptosis was detected by flow cytometry. The expression level of EHD2 was significantly increased in pLV-EHD2 group and decreased in pLV-siEHD2 group compared with control group and pLV-siEHD2 group, indicating the successfully established EHD2 overexpression cell line and EHD2 RNA interference cell line. EHD2 overexpression enhanced the proliferation, invasion, and migration but inhibited the apoptosis of ccRCC cells, while EHD2 interference showed opposite functions. EHD2 interference can inhibit the development of ccRCC by inhibiting the proliferation, invasion, and migration, and EHD2 can potentially serve as a molecular target for the clinical treatment of ccRCC.

scholarly journals FH3, A Domain Found in Formins, Targets the Fission Yeast Formin Fus1 to the Projection Tip During Conjugation

1998 ◽  
Vol 141 (5) ◽  
pp. 1217-1228 ◽  
Author(s):  
Janni Petersen ◽  
Olaf Nielsen ◽  
Richard Egel ◽  
Iain M. Hagan
Keyword(s):  
Fluorescent Protein ◽  
Protein Localization ◽  
Spindle Pole Body ◽  
Spindle Pole ◽  
Amino Terminal ◽  
Gfp Fusion Protein ◽  
Pole Body ◽  
Body Component ◽  
Green Fluorescent ◽  
Amino Terminal Domain

Formins are involved in diverse aspects of morphogenesis, and share two regions of homology: FH1 and FH2. We describe a new formin homology region, FH3. FH3 is an amino-terminal domain that differs from the Rho binding site identified in Bni1p and p140mDia. The Schizosaccharomyces pombe formin Fus1 is required for conjugation, and is localized to the projection tip in cells of mating pairs. We replaced genomic fus1+ with green fluorescent protein (GFP)- tagged versions that lacked either the FH1, FH2, or FH3 domain. Deletion of any FH domain essentially abolished mating. FH3, but neither FH1 nor FH2, was required for Fus1 localization. An FH3 domain–GFP fusion protein localized to the projection tips of mating pairs. Thus, the FH3 domain alone can direct protein localization. The FH3 domains of both Fus1 and the S. pombe cytokinesis formin Cdc12 were able to localize GFP to the spindle pole body in half of the late G2 cells in a vegetatively growing population. Expression of both FH3-GFP fusions also affected cytokinesis. Overexpression of the spindle pole body component Sad1 altered the distribution of both Sad1 and the FH3-GFP domain. Together these data suggest that proteins at multiple sites can interact with FH3 domains.

Sign in / Sign up

Export Citation Format

Share Document