Identification of reference genes for circulating microRNA analysis in colorectal cancer

Niu, Yanqin; Wu, Yike; Huang, Jinyong; Li, Qing; Kang, Kang; Qu, Junle; Li, Furong; Gou, Deming

doi:10.1038/srep35611

Cited by 52 publications

(54 citation statements)

References 42 publications

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“…33 34 In addition, studies are now also defining appropriate reference gene selection within tumour tissue and cancer cell lines. [35][36][37] Therefore, the aim of the present study was to define the impact of our FDG-MNPs in their non-magnetic stimulated state on tumour and non-tumour tissue in mice bearing MAC16 adenocarcinomas in a non-magnetic field environment. Here, we establish the expression profile of commonly used reference genes in a mouse adenocarcinoma model exposed to FDG-MNPs.…”

Section: Introductionmentioning

confidence: 99%

Tissue Morphology and Gene Expression Characterisation of Transplantable Adenocarcinoma Bearing Mice Exposed to Fluorodeoxyglucose-Conjugated Magnetic Nanoparticles

Watkins¹,

Pearce²,

Ünak³

et al. 2018

j biomed nanotechnol

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Fluorodeoxyglucose-conjugated magnetic nanoparticles, designed to target cancer cells with high specificity when heated by an alternating magnetic field, could provide a low-cost, non-toxic treatment for cancer. However, it is essential that the in vivo impacts of such technologies on both tumour and healthy tissues are characterised fully. Profiling tissue gene expression by semi-quantitative reverse transcriptase real-time PCR can provide a sensitive measurement of tissue response to treatment. However, the accuracy of such analyses is dependent on the selection of stable reference genes. In this study, we determined the impact of fluorodeoxyglucose-conjugated magnetic nanoparticles on tumour and nontumour tissue gene expression and morphology in MAC16 adenocarcinoma established male NMRI mice. Mice received an injection of 8 mg/kg body weight fluorodeoxyglucose-conjugated magnetic nanoparticles either intravenously in to the tail vein, directly into the tumour or subcutaneously directly overlying the tumour. Tissues from mice were sampled between 70 minutes and 12 hours post injection. Using the bioinformatic geNorm tool, we established the stability of six candidate reference genes (Hprt, Pgk1, Ppib, Sdha, Tbp and Tuba); we observed Pgk1 and Ppib to be the most stable. We then characterised the expression profiles of several apoptosis genes of interest in our adenocarcinoma samples, observing differential expression in response to mode of administration and exposure duration. Using histological assessment and fluorescent TUNNEL staining, we observed no detrimental impact on either tumour or non-tumour tissue morphology or levels of apoptosis. These observations define the underlying efficacy of fluorodeoxyglucose-conjugated magnetic nanoparticles on tumour and non-tumour tissue morphology and gene expression, setting the basis for future studies.

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Section: Introductionmentioning

confidence: 99%

Tissue Morphology and Gene Expression Characterisation of Transplantable Adenocarcinoma Bearing Mice Exposed to Fluorodeoxyglucose-Conjugated Magnetic Nanoparticles

Watkins¹,

Pearce²,

Ünak³

et al. 2018

j biomed nanotechnol

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show abstract

“…To determine the relative expression level of miR-145 and miR-143, three stem-loop primers (miR-145, 5 0 -CTC AAC TGG TGT CGT GGA GTC GGC AAT TCA GTT GAG AGG GAT TC-3 0 ; miR-143, 5 0 -GTC GTA TCC AGT GCT GGG TCC GAG TGA TTC GCA CTG GAT ACG ACT GAG CTA CA-3 0 ; and miR-93 [which has been identified as a reference gene for qPCR analysis of miRNA levels 46,47 ], 5 0 -CTC AAC GGT GTC GTG GAG TCG GCA ATT CAG TTG AGC TAC CTG C-3 0 ) were used for RT with an iScript cDNA synthesis kit (#1708890, Bio-Rad) according to the manufacturer's instructions. qPCR was then conducted using three primer pairs targeting miR-145 (forward, 5 0 -CGG CGG GTC CAG TTT TCC CAG G-3 0 ; reverse, 5 0 -CTG GTG TCG TGG AGT CGG CAA TTC-3 0 ), miR-143 (forward, 5 0 -CCT GGC CTG AGA TGA AGC AC-3 0 ; reverse, 5 0 -CAG TGC TGG GTC CGA GTG A-3 0 ), and miR-93 (forward, 5 0 -CGG CGG CAA AGT GCT GTT CGT G-3 0 ; reverse, 5 0 -CTG GTG TCG TGG AGT CGG CAA TTC-3 0 ).…”

Section: Quantitative Reverse Transcriptase Pcr (Qrt-pcr)mentioning

confidence: 99%

MicroRNA Modification of Coxsackievirus B3 Decreases Its Toxicity, while Retaining Oncolytic Potency against Lung Cancer

Liu

Xue

Deng

et al. 2020

Molecular Therapy - Oncolytics

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We recently discovered that coxsackievirus B3 (CVB3) is a potent oncolytic virus against KRAS mutant lung adenocarcinoma. Nevertheless, the evident toxicity restricts the use of wild-type (WT)-CVB3 for cancer therapy. The current study aims to engineer the CVB3 to decrease its toxicity and to extend our previous research to determine its safety and efficacy in treating TP53/RB1 mutant small-cell lung cancer (SCLC). A microRNA-modified CVB3 (miR-CVB3) was generated via inserting multiple copies of tumor-suppressive miR-145/miR-143 target sequences into the viral genome. In vitro experiments revealed that miR-CVB3 retained the ability to infect and lyse KRAS mutant lung adenocarcinoma and TP53/RB1mutant SCLC cells, but with a markedly reduced cytotoxicity toward cardiomyocytes. In vivo study using a TP53/RB1mutant SCLC xenograft model demonstrated that a single dose of miR-CVB3 via systemic administration resulted in a significant tumor regression. Most strikingly, mice treated with miR-CVB3 exhibited greatly attenuated cardiotoxicities and decreased viral titers compared to WT-CVB3-treated mice. Collectively, we generated a recombinant CVB3 that is powerful in destroying both KRAS mutant lung adenocarcinoma and TP53/RB1-mutant SCLC, with a negligible toxicity toward normal tissues. Future investigation is needed to address the issue of genome instability of miR-CVB3, which was observed in~40% of mice after a prolonged treatment.

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“…The relative miRNA expression was calculated using the Ct method and normalized on miR-93-5p. Several pieces of evidence reported high stability of miR-93-5p in bio uids [32][33][34][35] thus miR-93-5p was suggested as plasmatic reference gene in manufacture's handbook. According to this, the expression levels of miR-93-5p in plasma samples of our cohort showed comparable expression levels without signi cant difference among groups (data unshown).…”

Section: Analysis Of the Expression Of Circulating Mirnasmentioning

confidence: 99%

Apolipoprotein E polymorphism and oxidative stress in human peripheral blood cells: can physical activity reactivate the proteasome system through epigenetic mechanisms?

Piccarducci

Daniele

Polini

et al. 2020

Preprint

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Background Alzheimer’s Disease (AD) is characterised by proteasome activity impairment and oxidative stress, resulting in β-amyloid (Aβ) production/degradation imbalance. Apolipoprotein E (ApoE) is implicated in Aβ clearance, and ApoE ε4 isoform predisposes to AD development. Regular physical activity is known to reduce AD progression. However, the impact of ApoE polymorphism and physical exercise on Aβ production and proteasome system activity has never been investigated in human peripheral blood cells.Methods Healthy subjects were enrolled and classified based on the ApoE polymorphism (by the restriction fragment length polymorphism technique) and physical activity level (Borg Scale), dividing them in ApoE ε4/non-ε4 carriers and active/non-active subjects. The plasma antioxidant capability (AOC), the erythrocyte Aβ production/accumulation, and the nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated proteasome functionality were evaluated in all groups by chromatographic and immunoenzymatic assay, respectively. Moreover, epigenetic mechanisms were investigated considering the expression of the histone deacetylase 6, employing a competitive ELISA, and the modulation of two keys miRNAs (miR-153-3p and miR-195-5p), through miRNeasy Serum/Plasma Mini Kit.Results ApoE ε4 subjects showed a reduction in plasma AOC and an increase of the Nrf2 blocker, miR-153-3p, contributing to an enhancement of the erythrocyte concentration of Aβ. Physical exercise increased plasma AOC and reduced the amount of Aβ and its precursor, involving a reduced miR-153-3p expression and a miR-195-5p enhancement.Conclusions Our data highlight the impact of ApoE genotype on the amyloidogenic pathway and the proteasome system, and suggest the positive impact of physical exercise, also through epigenetic mechanisms.

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Identification of reference genes for circulating microRNA analysis in colorectal cancer

Cited by 52 publications

References 42 publications

Tissue Morphology and Gene Expression Characterisation of Transplantable Adenocarcinoma Bearing Mice Exposed to Fluorodeoxyglucose-Conjugated Magnetic Nanoparticles

Tissue Morphology and Gene Expression Characterisation of Transplantable Adenocarcinoma Bearing Mice Exposed to Fluorodeoxyglucose-Conjugated Magnetic Nanoparticles

MicroRNA Modification of Coxsackievirus B3 Decreases Its Toxicity, while Retaining Oncolytic Potency against Lung Cancer

Apolipoprotein E polymorphism and oxidative stress in human peripheral blood cells: can physical activity reactivate the proteasome system through epigenetic mechanisms?

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