scholarly journals MicroRNA-200c-3p Functions as a Tumor Suppressor in Kidney Renal Clear Cell Carcinoma by Targeting Vascular Endothelial Growth Factor A

2021 ◽  
Author(s):  
Yanmin Li ◽  
Hui Xu ◽  
Qianghai Wen ◽  
Xiaoqi Yan ◽  
Gang xu
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Clear Cell ◽  
Clear Cell Carcinoma ◽  
Renal Clear Cell Carcinoma ◽  
Luciferase Reporter ◽  
Vascular Endothelial ◽  
Reporter Assay ◽  
Wound Healing Assay ◽  
Endothelial Growth Factor ◽  
Dual Luciferase Reporter Assay

Abstract Background: MicroRNA(miR)-200c-3p is a tumor suppressor that helps inhibit the progression of various types of cancer. However, its role in kidney renal clear cell carcinoma (KRIC) is unknown. The purpose of this study was to explore the biological function and regulatory mechanism of miR-200c-3p in the development of KRIC.Methods: The relative levels of miR-200c-3p and vascular endothelial growth factor A (VEGFA) in KIRC tissues and cells were determined using the qRT-PCR technique. Transwell and wound healing assay methods were used to understand the effect of miR-200c-3p on the migration and invasion of 786-O and Caki-1 cells. The ability of miR-200c-3p to target VEGFA was determined using the Dual-Luciferase reporter assay system Results: MiR-200c-3p was downregulated in KIRC tissues and cell lines. The overexpression of miR-200c-3p attenuated the migratory capacities of 786-O and Caki-1 cells. The migratory ability increased under conditions of miR-200c-3p knockdown. Analysis of the results obtained using the wound healing assay revealed that overexpression of miR-200c-3p reduced the migratory abilities of Caki-1 and 786-O cells, and inhibition of miR-200c-3p accelerated the process of wound closure. Analysis of the results obtained from screening tests and by conducting the Dual-Luciferase reporter assay revealed that VEGFA was the direct target of miR-200c-3p. The VEGFA mRNA level was low in the 786-O and Caki-1 cells under conditions of overexpressed miR-200c-3p. High levels of VEGFA mRNA were observed in the cells post miR-200c-3p knockdon.Conclusions: The tumor suppression ability of miR-200c-3p in KIRC can be achieved by regulating the expression of VEGFA.

Vascular Endothelial Growth Factor (VEGF) Polymorphisms and Serum VEGF Levels in Women With Epithelial Ovarian Cancer, Benign Tumors, and Healthy Ovaries

2017 ◽  
Vol 27 (6) ◽  
pp. 1088-1095 ◽  
Author(s):  
Blanca González-Palomares ◽  
Pluvio J. Coronado Martín ◽  
María Luisa Maestro de las Casas ◽  
Silvia Veganzones de Castro ◽  
Sara Rafael Fernández ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Clear Cell ◽  
Clear Cell Carcinoma ◽  
Serum Levels ◽  
Benign Tumors ◽  
Vascular Endothelial ◽  
Endothelial Growth Factor ◽  
Worse Prognosis

ObjectiveThis study analyzed the relation of 5 single-nucleotide polymorphisms (SNPs) in the VEGF (vascular endothelial growth factor) gene in patients with epithelial ovarian cancer (EOC), compared with patients carrying benign tumors or healthy ovaries. We studied serum VEGF levels and the relation with SNPs and association between VEGF SNPs and haplotypes with progression-free survival (PFS) in patients with cancer.MethodsThe genotyping of VEGF gene polymorphisms (−2578 C/A, −1154 G/A, −460 T/C, +405 G/C, +936 C/T) was performed in DNA isolated from blood samples of 100 women. The different genotypes were evaluated by quantitative real-time polymerase chain reaction. Vascular endothelial growth factor protein concentration was assessed in serum using solid-phase sandwich enzyme-linked immunosorbent assay.ResultsWe found statistically significant differences in the distribution of VEGF genotypes among the 3 groups of patients: −2578 C/A between those with EOC and healthy ovary (P = 0.04), −460 T/C between those with EOC and healthy ovary (P = 0.03), and −460 T/C between those with benign tumors and healthy ovary (P = 0.02). Vascular endothelial growth factor serum levels were analyzed in patients with EOC. Higher levels were found in patients with clear cell carcinoma compared with those with serous, mucinous, or endometrioid tumors (P < 0.05). No clear association was observed between VEGF SNPs and serum VEGF levels. There was no significant correlation between VEGF SNPs and PFS. In haplotype analysis, CGTCT and CGTGT showed worse prognosis without reaching the statistical significance. CGCGC and AGTGC haplotypes had statistically significant differences among patients with EOC, benign tumors, and healthy ovaries (Ps = 0.046 and 0.041, respectively).ConclusionsThe distribution of VEGF genotypes was different in patients with EOC, compared with those with benign tumors or women with healthy ovaries. Vascular endothelial growth factor serum levels were higher in patients with clear cell carcinoma. No correlation was found with improved PFS, but CGTCT and CGTGT haplotypes showed worse prognosis.

Investigation of Targeting Relationship between Micro-Rna-22 and Vegfr3 in Lung Squamous Cell Carcinoma

2020 ◽  
Vol 23 ◽  
Author(s):  
Zheng Dong ◽  
Qing-Hua Xu ◽  
Yuan-Bin Zhu ◽  
Yong-Feng Wang ◽  
Jie Xiong ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Lung Squamous Cell Carcinoma ◽  
Luciferase Reporter ◽  
Control Group ◽  
Vascular Endothelial ◽  
Luciferase Reporter Gene ◽  
Endothelial Growth Factor

Aims : The present study explored the clinical significance of microRNA-22 (miR-22) expression in lung squamous cell carcinoma and to explore the targeting relationship with vascular endothelial growth factor receptor 3 (VEGFR3). Methods: A total of 49 patients with lung squamous cell carcinoma who underwent surgical treatment was selected. The expression of miR-22 was detected by fluorescence quantitative real-time PCR (qPCR), the expression of VEGFR3 was detected by Western blotting assays, and D240 labeled microlymphatic vessels density (MLVD) was detected immunohistochemistry (IHC). Lung squamous cell carcinoma cell line SK-MES-1 was selected and the targeting relationship between miR-22 and VEGFR3 was analyzed by double luciferase reporter gene assay. Western blotting assays were used to detect the expression of vascular endothelial growth factor-D (VEGF-D) and D240 in the blank control group, empty vector transfection group, miR-22 transfection group, miR-22 and VEGFR3 co-transfection group. Results: The expression range of miR-22 in lung squamous cell carcinoma was 0.8-3.5. The expression of miR-22 in lung squamous cell carcinoma was significantly different by tumor maximum diameter, lymph node metastasis, vascular invasion and TNM stage. The expression of miR-22 was linked to survival time. There was a negative correlation between miR-22 and VEGFR3, miR-22 and MLVD. Double luciferase reporter gene assays showed that miR-22 reduced the luciferase activity of pGL3-VEGFR3-WT transfected cells. Compared with the control group, the expression of VEGF-D and D2-40 in the miR-22 transfection group was significantly decreased. However, VEGF-D and D240 in the miR-22 and VEGFR3 cotransfection group reversed the changes. Conclusion: We assumed that the abnormal expression of miR-22 in lung squamous cell carcinoma may be involved in the development and progression of lung squamous cell carcinoma. MiR-22 negatively regulated the target gene VEGFR3 to mediate lymphangiogenesis. The expression of miR-22 may also be linked to the prognosis of the disease.

CD40-induced transcriptional activation of vascular endothelial growth factor involves a 68-bp region of the promoter containing a CpG island

2004 ◽  
Vol 287 (3) ◽  
pp. F512-F520 ◽  
Author(s):  
Peter H. Lapchak ◽  
Michael Melter ◽  
Soumitro Pal ◽  
Jesse A. Flaxenburg ◽  
Christopher Geehan ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Transcriptional Activation ◽  
Cpg Island ◽  
Transcriptional Repressor ◽  
Renal Physiology ◽  
Luciferase Reporter ◽  
Vascular Endothelial ◽  
Cpg Dinucleotides ◽  
Endothelial Growth Factor

Vascular endothelial growth factor (VEGF) is produced by several cell types in the kidney, and its expression is tightly regulated for the maintenance of normal renal physiology. Increases or decreases in its expression are associated with proteinuria and renal disease. Recently, we found that the expression of VEGF is markedly induced following interactions between CD40 ligand (CD40L) and CD40. Here, endothelial cells (EC) or Jurkat T cell lines were transiently transfected with luciferase reporter constructs under the control of the human VEGF promoter and were treated with human soluble CD40L (sCD40L). We identified a CD40-responsive 68-bp region (bp −50 to +18) of the promoter and 43 bp within this region (bp −25 to +18) that have 97% homology to a sequence of CpG dinucleotides. A computerized search revealed that the CpG region has putative binding domains for the transcriptional repressor protein methyl CpG binding protein-2 (MeCP2). In EMSA, we found that the 43-bp methylated sequence formed four complex(es) with nuclear extracts from untreated EC and reduced binding of at least one complex when nuclear lysates from sCD40L-activated EC (30 min) were used. Supershift analysis using anti-MeCP2 demonstrated that most of the complex(es) in both untreated and sCD40L-activated EC involved interactions between the 43-bp DNA and MeCP2. In addition, we found that other CpG binding proteins may also interact with this region of the promoter. Taken together, this is the first demonstration that CpG binding transcriptional repressor proteins including MeCP2 may be of importance in VEGF biology.

scholarly journals Prognostic Value of Vascular Endothelial Growth Factor A in the Prediction of the Tumor Aggressiveness in Clear Cell Renal Cell Carcinoma

2017 ◽  
Vol 5 (2) ◽  
pp. 167-172 ◽  
Author(s):  
Fahredin Veselaj ◽  
Suzana Manxhuka-Kerliu ◽  
Arber Neziri ◽  
Labinot Shahini ◽  
Shefki Xharra ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Renal Cell ◽  
Prognostic Value ◽  
Clear Cell ◽  
Vascular Endothelial ◽  
Cell Renal Cell Carcinoma ◽  
Free Survival ◽  
Endothelial Growth Factor

BACKGROUND: Clear cell renal cell carcinoma (CCRCC) is the most predominant renal tumour with unpredictable tumour behaviour. The aim of the study is to investigate the prognostic value of vascular endothelial growth factor A (VEGF-A) expression in CCRCC and to correlate it with other histological parameters as well as with patient's survival.MATERIAL AND METHODS: Tumour blocks were taken from 40 patients with histopathology diagnosis of CCRCC and tissue block from 20 normal kidneys as a control group were examined using the immuno-histochemical staining for VEGF-A.RESULTS: The VEGF A expression in CCRCC was significantly higher than in the normal kidney tissues (U’ = 720, P < 0.0001). VEGF A expression values in CCRCC were positively correlated with Disease Free Survival (r = 0.335, P = 0.034) and the tumor necrosis degree (r = 0.181, P = 0.262). VEGF-A expression values in CCRCC did not correlate with CD 31 expression (r = -0.09, P = 0.549), and Progression Free Survival (r = -0.07, P = 0.838). VEGF A expression values in CCRCC were negatively correlated with the tumor nuclear grade (r = -0.161, P = 0.318); the pathological tumor stage (r = -0.371, P = 0.018); the tumor size (r = -0.361, P = 0.022); the degree of tumor hemorrhage (r = -0.235, P = 0.143); and Cancer Specific Survival   (r = -0.207, P = 0.713).CONCLUSIONS: VEGF-A expression can be used to stratify advanced and metastatic CCRCC patients into low-benefit and high-benefit groups. Based on this study outcome it would be useful to perform IHC staining for VEGF-A expression in all patients with advanced and metastatic CCRCC.

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