scholarly journals Identification of Appropriate Housekeeping Genes for Gene Expression Studies in Human Renal Cell Carcinoma Under Hypoxic Conditions

2021 ◽  
Author(s):  
Luiz Felipe S. Teixeira ◽  
Rodrigo Gigliotti ◽  
Luana da Silva Ferreira ◽  
MARIA Helena Bellini
Keyword(s):  
Gene Expression ◽  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Expression Analysis ◽  
Renal Cell ◽  
Gene Expression Analysis ◽  
Housekeeping Genes ◽  
Human Renal Cell Carcinoma ◽  
Von Hippel Lindau ◽  
Hypoxic Conditions

Abstract Background: Due to the loss of von Hippel-Lindau tumor suppressor function, clear renal cell carcinoma (ccRCC) deregulates hypoxia pathways. Quantitative PCR is a powerful tool for quantifying differential expression between normal and cancer cells. Reliable gene expression analysis requires the use of genes encoding housekeeping genes. Therefore, in this study, eight reference candidate genes were evaluated to determine their stability in 786-0 cells under normoxic and hypoxic conditions. Methods and Results: Four different tools were used to rank the most stable genes: GeNorm, NormFinder, BestKeeper, and Comparative Ct (ΔCt), and a general ranking was performed using the RankAggreg. According to the four algorithms, the TFRC reference gene was identified as the most stable, and therefore, no agreement was observed for the 2nd and 3rd positions. A general classification was then established using the RankAggreg tool. Finally, the three most suitable reference genes to be used in 786-0 cells under normoxic and hypoxic conditions were TFRC, RPLP0, and SDHA. Conclusions: To our knowledge, this is the first study to evaluate reliable genes that can be used in gene expression analysis in ccRcc under a hypoxic environment.

scholarly journals Correlation analysis of VHL and Jade-1 gene expression in human renal cell carcinoma

Open Medicine ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. 226-230 ◽  
Author(s):  
Wu Xiao-fen ◽  
Chen Ting ◽  
Li Jie ◽  
Ma Deng-yang ◽  
Zhu Qing-feng ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Tumor Grade ◽  
Human Renal Cell Carcinoma ◽  
Clinical Prognosis ◽  
Phd Finger ◽  
Von Hippel Lindau ◽  
Normal Tissues

AbstractObjectiveThe aim of this study was to investigate the correlation of von Hippel-Lindau tumor suppressor (VHL) mRNA expression and jade family PHD finger 1 (Jade-1) gene expression in patients with renal cell carcinoma (RCC). Another aim of this study was to analyze the relationship of these two genes with clinicalpathological features of the RCC patients. Methods: A total of 75 RCC patients who received surgically therapy in our hospital were included. All patients had complete pathological data. The expression of VHL/Jade-1 was determined by real-time polymerase chain reaction (RT-PCR). Results: VHL and Jade-1 were both obviously downregulated in RCC tissues than that of the matched normal tissues, and both negatively correlated with tumor size as well as tumor grade. And we found a fine association of VHL gene expression with Jade-1. Conclusion: VHL/Jade-1 exhibited significantly decreased expression in RCC tissues and was closely related to the clinical prognosis of patients. The finding of VHL expression positively correlated with Jade-1 expression shed light and provided crucial evidence on the connection of VHL protein with Wnt/b-catenin pathway.

Markers of sunitinib-resistance in clear cell renal cell carcinoma: A gene expression analysis.

2014 ◽  
Vol 32 (4_suppl) ◽  
pp. 533-533
Author(s):  
Òscar Reig ◽  
Mercedes Marín-Aguilera ◽  
Juan José Lozano ◽  
Blanca Gonzalez ◽  
Carme Mallofré ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Expression Analysis ◽  
Renal Cell ◽  
Gene Expression Analysis ◽  
Predictive Biomarkers ◽  
Intrinsic Resistance ◽  
Primary Tumors ◽  
Novel Biomarkers

533 Background: Sunitinib is a standard first line treatment of metastatic renal cell carcinoma (ccRCC). Approximately 20% of patients experience primary resistance to therapy and no predictive biomarkers are available. The aim of our study is to discover novel biomarkers to predict response to sunitinib and to generate hypothesis about mechanisms of intrinsic resistance. Methods: Gene expression analysis was performed in formalin-fixed paraffin embedded (FFPE) samples from 44 patients with metastatic ccRCC treated with sunitinib in our institution. Affymetrix Human Gene 2.0 ST array was performed in primary tumors from 6 extremely sensitive (progression free survival (PFS) > 24 months) and 8 refractory (progression disease as best response) ccRCC. Technical validation with qPCR (Fluidigm Dynamic 96.96 Array) was performed in the whole cohort. The ΔΔCt method was used to quantify the relative amount of mRNA. Clinical and pathological data were correlated with gene expression. Results: 330 genes were differentially expressed between refractory and sensitive patients (p≤0.05). Network analysis showed 16 significant networks represented. Gene expression of 96 selected markers was tested in 47 primary tumors and 24 metastases. We found IL8, VEGF and NOTCH pathways upregulated on refractory patients. Survival analysis showed that the overexpression of IL8 correlated with a worse PFS (HR: 2.42, 95%CI1.27 – 4.63, p=0.0075) and overall survival (OS) (HR: 3.42, 95%CI 1.50 – 7.79, p=0.0034). Overexpression of VEGFBcorrelated with a prolonged PFS (HR 0.52, 95% CI 0.28 – 0.98, p=0.0415). Conclusions: Our results confirm the predictive value of IL8 expression in a cohort of ccRCC patients treated with sunitinib and suggests novel biomarkers of response to sunitinib. These data will be further validated in an independent cohort of patients.

scholarly journals In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR

2007 ◽  
Vol 8 (1) ◽  
pp. 47 ◽  
Author(s):  
Monika Jung ◽  
Azizbek Ramankulov ◽  
Jan Roigas ◽  
Manfred Johannsen ◽  
Martin Ringsdorf ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Real Time Pcr ◽  
Renal Cell ◽  
Reference Genes ◽  
Human Renal Cell Carcinoma ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference
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