Identification of reliable reference genes for gene expression studies in mouse models under microplastics stress

Mu, Ju; Wang, Yu; Wang, Miao; Zhang, Dayi; Liu, Mingying

doi:10.1016/j.ecoenv.2023.114569

Cited by 11 publications

(4 citation statements)

References 69 publications

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“…In this study, to obtain reliable results, we applied four algorithms: ΔCt method, BestKeeper, NormFinder, and geNorm to identify reliable reference genes for hiPSCs. Reference gene validation based on these four algorithms has been reported in several studies on various cell types and experimental conditions [14][15][16][17] .…”

Section: Discussionmentioning

confidence: 99%

Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human-induced pluripotent stem cells

Okamoto,

Inagaki,

Okamura

et al. 2024

Preprint

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Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is an essential tool for gene expression analysis; however, choosing appropriate reference genes for normalization is crucial to ensure data reliability. Most studies on osteogenic differentiation have had limited success in identifying optimal reference genes; to the best of our knowledge, no optimal reference genes in three-dimensional (3D) osteogenic differentiation culture experiments using human induced pluripotent stem cells (hiPSCs) have been identified. In this study, we aimed to identify stable reference genes that could be used for normalization in gene expression analyses during the 3D osteogenic differentiation of hiPSCs using an atelocollagen sponge as the scaffold. Four algorithms—ΔCt, BestKeeper, NormFinder, and geNorm—were used to evaluate the stability of 14 candidate reference genes. TATA box-binding protein, hypoxanthine phosphoribosyltransferase 1, and 14-3-3 protein zeta polypeptide emerged as the most stable reference genes. In comparison, conventionally used reference genes (beta-2 microglobulin and beta-actin) ranked among those with low stability. We also demonstrated the successful 3D osteogenic differentiation of hiPSCs on the atelocollagen sponge. Our findings provide valuable insights into reference gene selection and bone tissue regeneration from hiPSCs, which will improve the treatment prospects for bone defects and other similar conditions in regenerative medicine.

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Section: Discussionmentioning

confidence: 99%

Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human-induced pluripotent stem cells

Okamoto,

Inagaki,

Okamura

et al. 2024

Preprint

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“…However, another study showed that serum CRE decreased in mice after MP treatment, while BUN was not affected [22]. Xiong et al reported the vacuolar and granular degeneration of renal tubular epithelial cells and tubular atrophy in mice kidneys after exposure to MPs [25], while no obvious damage in kidney tissues was presented in Mu et al's study [24]. These disparities could mainly be attributed to varying MP exposure dosages.…”

Section: Introductionmentioning

confidence: 94%

“…Previous studies have documented the bioaccumulation of MPs with varying particle sizes in the kidneys of mice after oral administration [21]. Although a few studies have found that MPs could induce histopathological and functional abnormalities regarding the kidney following gastrointestinal exposure, the results are inconsistent [22][23][24][25]. For example, Wang et al discovered that polystyrene microplastics (PS-MPs) could lead to lower renal coefficients and higher levels of serum creatinine (CRE) and blood urea nitrogen (BUN) in rats [23].…”

Section: Introductionmentioning

confidence: 99%

Effects of Microplastic (MP) Exposure at Environmentally Relevant Doses on the Structure, Function, and Transcriptome of the Kidney in Mice

Shen,

Zhang,

Wang

et al. 2023

Molecules

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As a common emerging environmental pollutant, microplastics (MPs) have been detected in a variety of environmental media and human bodies. The potential toxic effects and mechanisms of MPs need to be revealed urgently. MPs can be deposited in the kidney, and exposure to high doses of MPs can cause nephrotoxicity in experimental animals. In this study, we investigated the effects of exposure to polystyrene microplastics (PS-MPs) at environmentally relevant doses (0.1 and 1 mg/L) on kidney structure, function, and transcriptome in mice. We found that mice exposed to PS-MPs in drinking water for eight weeks had no change in body weight or kidney coefficient. PS-MPs administration decreased the levels of blood urea nitrogen (BUN) in mice, while serum creatinine (CRE) and uric acid (UA) concentrations were unaffected. Through using periodic acid–Schiff (PAS) and Masson staining, we discovered that the glomerular tuft area increased in the PS-MP-treated mice, while the degree of renal fibrosis remained unchanged. Furthermore, renal cortex transcriptomic analysis identified 388 and 303 differentially expressed genes (DEGs) in the 0.1 and 1 mg/L dose groups, respectively. The DEGs were highly enriched in mitochondrial-related terms and pathways of thermogenesis and oxidative phosphorylation. Moreover, protein–protein interaction (PPI) network analysis revealed that cytochrome b-c1 complex subunit 10 (UQCR11) and cytochrome c oxidase subunit 3 (MT-CO3) were important node proteins. These findings suggest that environmental exposure to MPs can cause abnormalities in renal structure and filtration function and that long-term exposure to MPs may be a risk factor for renal disease.

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“…The use of (RT-PCR) to assess the mRNA levels of the target genes needs to use the housekeeping genes such as β-actin (9). It is presumed that the expression of these genes keeps constant normalizing the variations in signal estimation and processing (10).…”

Section: Introductionmentioning

confidence: 99%

Role Toll-like receptor 4 gene underlying Hypoxia-inducible transcription factor 1α gene expression in cervical cancer of women

Nama¹,

Kraidi²

2023

Preprint

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Molecular methods are being used in cervical cancer screening to identify high-risk human papillomavirus. Annually, 490,000 new infections are diagnosed with papillomavirus, and about 298,000 death cases due to cervical cancer worldwide. Pap and biopsy samples from 65 samples from cervixes were collected from women with histological changes from Alsider Teaching Hospital, Maysan Hospital for Children, South of Iraq. The viruses were detected by RT-PCR in 69.2 % of the all cases. Extracted RNA samples were used to analyze the gene expression of Toll-like receptor 4 and hypoxia-inducible transcription factor 1α from biopsy samples. It was found that the number of positive samples for viruse genotype 16 was in 30 samples, which represents 66.6 %, while the genotype 18 was detected in 17 samples, which represents 37.7%, while 9 samples gave positive results for genotype 31 and represented about 20 %, relying on E6 or E7 specific primers in cDNA template by real-time PCR ,and the values of Toll-like receptor 4 in negative samples were normalized to the housekeeping gene (β actin) to 1 and its values in positive samples were compared to that 1 which was more roughly in 1-fold, and the hypoxia-inducible transcription factor 1α gene expression in positive samples was more in roughly 48-fold compared to its value in the housekeeping gene (β actin) to 1 (negative control). The study concluded that toll-like receptor 4 and hypoxia-inducible transcription factor 1α are overexpressed in cervical cancer, and both synergistically promote the development of cervical cancer.

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Identification of reliable reference genes for gene expression studies in mouse models under microplastics stress

Cited by 11 publications

References 69 publications

Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human-induced pluripotent stem cells

Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human-induced pluripotent stem cells

Effects of Microplastic (MP) Exposure at Environmentally Relevant Doses on the Structure, Function, and Transcriptome of the Kidney in Mice

Role Toll-like receptor 4 gene underlying Hypoxia-inducible transcription factor 1α gene expression in cervical cancer of women

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