GAPDH and PUM1: Optimal Housekeeping Genes for Quantitative Polymerase Chain Reaction-Based Analysis of Cancer Stem Cells and Epithelial-Mesenchymal Transition Gene Expression in Rectal Tumors

Vermani, Litika; Kumar, Rajeev; Kumar, Nachimuthu Senthil

doi:10.7759/cureus.12020

Cited by 15 publications

(15 citation statements)

References 28 publications

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“…Recurrence of epithelial ovarian cancer is seriously deteriorated with little chance of complete cure. Therefore, the treatment of recurrent epithelial ovarian cancer is mainly chemotherapy to control the disease condition, improve the existing clinical symptoms and quality of life of patients, and prolong the survival time of patients [ 22 ]. According to the clinical effect after treatment, bevacizumab combined with chemotherapy has better therapeutic effect.…”

Section: Discussionmentioning

confidence: 99%

Effect of bevacizumab combined with chemotherapy on SDF-1 and CXCR4 in epithelial ovarian cancer and its prognosis

2022

World J Surg Onc

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Background The effect of bevacizumab combined with chemotherapy on the expression of stromal cell-derived factor-1 (SDF-1) and receptor CXCR4 in epithelial ovarian cancer tumor cells and its prognosis are unknown. Our work aimed to investigate the effect of chemotherapy +/− bevacizumab on these markers and the impact of this treatment modality in clinical outcomes. Methods Altogether 68 patients with epithelial ovarian cancer who were treated with chemotherapy in our hospital from June 2018 to June 2019 were selected. It was an open-labeled and controlled clinical trial (ethical approval no. 20180435). The patients were grouped according to their admission order. Patients treated with paclitaxel and carboplatin were included in group A, while patients treated with bevacizumab, paclitaxel, and carboplatin were included in group B. qRT-PCR was used to detect the changes of SDF-1 and CXCR4 before and after chemotherapy. Various clinical indicators of patients in the two groups were recorded to analyze the clinical efficacy, and safety of different treatment modalities and the prognosis of the two groups was analyzed. Results The relative expression of SDF-1 and CXCR4 was positively correlated with epithelial ovarian cancer stages (P<0.00). Together, SDF-1 and CXCR4 were positively correlated in epithelial ovarian cancer staging (P<0.001). SDF-1 and CXCR4 in both groups after chemotherapy were significantly decreased (P<0.001), and the downregulation of SDF-1 and CXCR4 expression in group B was significantly higher than that in group A after chemotherapy (P<0.001). No significant difference in the metastasis rates of the two groups before chemotherapy was observed (P>0.05), but the recurrence rate after 1 year was lower in group B than in group A (P<0.05). Conclusion Adding bevacizumab diminished the expression of related cancer markers SDF-1 and CXCR4 more than chemotherapy alone in patients with epithelial ovarian cancer. Furthermore, better rates of recurrence with no concerns regarding adverse drug reactions or quality of life were seen in bevacizumab plus chemotherapy group.

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Section: Discussionmentioning

confidence: 99%

Effect of bevacizumab combined with chemotherapy on SDF-1 and CXCR4 in epithelial ovarian cancer and its prognosis

2022

World J Surg Onc

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“…The expression level of GAPDH might be affected by the disease status of septic patients. The GAPDH gene was highly conserved(28) and was often used as a reference gene for RT-qPCR (29)(30)(31). GAPDH was an important enzyme in glycolysis, which catalyzed the oxidative dehydrogenation and phosphorylation of glyceraldehyde 3-phosphate to produce 1,3-diphosphoglyceric acid.…”

Section: Discussionmentioning

confidence: 99%

Validation of reference genes for the normalization of the RT- qPCR in peripheral blood mononuclear cells of septic patients

Song

et al. 2022

Preprint

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Objective To screen and validate reference genes suitable for gene mRNA expression study in peripheral blood mononuclear cells (PBMCs) between septic patients and healthy controls (HC). Methods Total RNA in PBMCs was extracted and RT-qPCR was used to determine the mRNA expression profiles of 9 candidate genes, including ACTB, B2M, GAPDH, GUSB, HPRT1, PGK1, RPL13A, SDHA and YWHAZ. The genes expression stabilities were assessed by both geNorm and NormFinder software. Results YWHAZ was the most stable gene among the 9 candidate genes evaluated by both geNorm and NormFinder in mixed and sepsis groups. The most stable gene combination in mixed group analyzed by geNorm was the combination of GAPDH, PKG1 and YWHAZ, while that in sepsis group was the combination of ACTB, PKG1 and YWHAZ. Conclusion Some commonly used stably expressed reference genes might be up-regulated or down-regulated in septic patients compared with healthy people. Of all nine genes studied, YWHAZ was found to be the best reference gene for studying the PBMC gene expression in septic patients. The best reference gene combination for septic patients’ investigation was ACTB, PKG1 and YWHAZ. We provide an important data for selecting the most stable reference genes to standardize RT-qPCR data, gene expression verification of sepsis and future gene function exploration.

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“…The three cecum samples (one from a normal group and two from a tumorous group) were excluded from the analyses. qRT-PCR was performed with FastStart Green PCR Master Mix, and gene expression levels were normalized to the GAPDH and Pum1 levels (25)based on the method of Hellemans and colleagues (26). Database analyses were performed using GENT2 database (27) and The Cancer Genome Atlas Colon Adenocarcinoma (TCGA-COAD) using the Xena Platform (28).…”

Section: Methodsmentioning

confidence: 99%

“…The Colon Cancer Tissue cDNA Array IV panel (HCRT104) was purchased from Origene Technologies (Origene Technologies Inc., Rockville, MD to the GAPDH and Pum1 levels [13]based on the method of Hellemans and colleagues [14]. Database analyses were performed using Gent2 database [15] and…”

Section: Tissue Cdna Array and Database Analysesmentioning

confidence: 99%

Exploring the Interdependence of TKS4 and CD2AP: Implications for EMT Process and Early Detection in Colon Cancer

Kurilla

László

Takács

et al. 2023

Preprint

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Colon cancer is a leading cause of death worldwide. Although several molecular biomarkers have been developed for colon cancer, more sensitive and specific methods are needed for its early detection. The TKS4 and CD2AP scaffold proteins have been linked to dynamic actin assembly-related processes, such as cell migration and epithelial-mesenchymal transition (EMT), although their co-instructive role during colon cancer development remains unknown. Therefore, this study was designed to investigate the interdependence of TKS4 and CD2AP during colon cancer progression as well as their involvement in EMT. This is the first study demonstrating a TKS4-CD2AP protein-protein interaction in vitro and in vivo. We describe that CD2AP and TKS4 interdependently induce a mesenchymal transition-like process and impair the migration ability of HCT116 colon cancer cells. Here we show that CD2AP is a novel TKS4 interacting partner, and together they form a scaffold complex that potentially regulates EMT-related signalling. Based on our analysis of the relative TKS4 and CD2AP expression levels, we propose that their levels might cooperatively serve as a reliable biomarker for colon cancer detection.

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GAPDH and PUM1: Optimal Housekeeping Genes for Quantitative Polymerase Chain Reaction-Based Analysis of Cancer Stem Cells and Epithelial-Mesenchymal Transition Gene Expression in Rectal Tumors

Cited by 15 publications

References 28 publications

Effect of bevacizumab combined with chemotherapy on SDF-1 and CXCR4 in epithelial ovarian cancer and its prognosis

Effect of bevacizumab combined with chemotherapy on SDF-1 and CXCR4 in epithelial ovarian cancer and its prognosis

Validation of reference genes for the normalization of the RT- qPCR in peripheral blood mononuclear cells of septic patients

Exploring the Interdependence of TKS4 and CD2AP: Implications for EMT Process and Early Detection in Colon Cancer

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