MicroRNA Isolation by Trizol-Based Method and Its Stability in Stored Serum and cDNA Derivatives

Trakunram, Keson; Champoochana, Nidanut; Chaniad, Pichitpon; Thongsuksai, Paramee; Raungrut, Pritsana

doi:10.31557/apjcp.2019.20.6.1641

Cited by 23 publications

(19 citation statements)

References 28 publications

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“…Their impact on series of cell signaling pathways related to tumorigenesis creates a wide research field towards their use as diagnostic and prognostic (pre)cancer biomarkers [ 73 , 100 , 118 , 140 , 163 ]. miRNAs are relatively easy to measure under laboratory conditions due to the much greater stability of the molecules compared to other RNA fractions [ 168 ]. However, the techniques of isolation, determination, normalization, and analysis of results require standardization in order to obtain repeatability and comparability of test results obtained in various research centers.…”

Section: Discussionmentioning

confidence: 99%

MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer

Pisarska

Baldy-Chudzik

2020

JCM

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The regulatory functions of microRNA (miRNA) are involved in all processes contributing to carcinogenesis and response to viral infections. Cervical cancer in most cases is caused by the persistence of high-risk human papillomavirus (HR-HPV) infection. While oncogenic human papillomaviruses induce aberrant expression of many cellular miRNAs, this dysregulation could be harnessed as a marker in early diagnosis of HR-HPV infection, cervical squamous intraepithelial lesions, and cancer. In recent years, growing data indicate that miRNAs show specific patterns at various stages of cervical pathology. The aim of this review is to systematize current reports on miRNA capacity that can be utilized in personalized diagnostics of cervical precancerous and cancerous lesions. The analysis of the resources available in online databases (National Center for Biotechnology Information—NCBI, PubMed, ScienceDirect, Scopus) was performed. To date, no standardized diagnostic algorithm using the miRNA pattern in cervical pathology has been defined. However, the high sensitivity and specificity of the reported assays gives hope for the development of non-invasive diagnostic tests that take into account the heterogeneity of tumor-related changes. Due to this variability resulting in difficult to predict clinical outcomes, precise molecular tools are needed to improve the diagnostic and therapeutic process.

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Section: Discussionmentioning

confidence: 99%

MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer

Pisarska

Baldy-Chudzik

2020

JCM

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“…Total RNA was isolated from breast milk (F0 and F1 generations) and liver and gonad tissues (F2 generation) using TRIZOL (Roche, Germany; Trakunram et al, 2019 ). Concentration and purity of RNA for each sample were confirmed using a Nanodrop1000 (Thermo Fisher Scientific, MA, United States).…”

Section: Methodsmentioning

confidence: 99%

Epigenetic Programming Through Breast Milk and Its Impact on Milk-Siblings Mating

et al. 2020

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Background The epigenetic effects of transmission of certain regulatory molecules, such as miRNAs, through maternal milk on future generations, are still unknown and have not been fully understood yet. We hypothesized that breastfeeding regularly by adoptive-mother may cause transmission of miRNAs as epigenetic regulating factors to the infant, and the marriage of milk-siblings may cause various pathologies in the future generations. Results A cross-fostering model using a/a and A vy /a mice had been established. F2 milk-sibling and F2 control groups were obtained from mating of milk-siblings or unrelated mice. Randomized selected animals in the both F2 groups were sacrificed for miRNA expression studies and the remainings were followed for phenotypic changes (coat color, obesity, hyperglycemia, liver pathology, and life span). The lifespan in the F2 milk-sibling group was shorter than the control group (387 vs 590 days, p = 0.011) and they were more obese during the aging period. Histopathological examination of liver tissues revealed abnormal findings in F2 milk-sibling group. In order to understand the epigenetic mechanisms leading to these phenotypic changes, we analyzed miRNA expression differences between offspring of milk-sibling and control matings and focused on the signaling pathways regulating lifespan and metabolism. Bioinformatic analysis demonstrated that differentially expressed miRNAs were associated with pathways regulating metabolism, survival, and cancer development such as the PI3K-Akt, ErbB, mTOR, and MAPK, insulin signaling pathways. We further analyzed the expression patterns of miR-186-5p, miR-141-3p, miR-345-5p, and miR-34c-5p and their candidate target genes Mapk8, Gsk3b, and Ppargc1a in ovarian and liver tissues. Conclusion Our findings support for the first time that the factors modifying the epigenetic mechanisms may be transmitted by breast milk and these epigenetic interactions may be transferred transgenerationally. Results also suggested hereditary epigenetic effects of cross-fostering on future generations and the impact of mother-infant dyad on epigenetic programming.

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“…Peripheral blood (5 ml) was collected in a clotting tube (Greiner Bio-One, Kremsmünster, Austria). Serum was isolated by centrifugation and prepared for miRNA isolation, as previously described [ 8 ].…”

Section: Methodsmentioning

confidence: 99%

Serum miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer

et al. 2020

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Tumor-promoting cytokines are a cause of tumor progression; therefore, identifying key regulatory microRNAs (miRNAs) for controlling their production is important. The aim of this study is to identify promising miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer (NSCLC). We identified circulating miRNAs from 16 published miRNA profiles. The selected miRNAs were validated in the serum of 32 NSCLC patients and compared with 33 patients with other lung diseases and 23 healthy persons using quantitative real-time PCR. The cytokine concentration was investigated using the enzyme-linked immunoassay in the same sample set, with clinical validation of the miRNAs. The correlation between miRNA expression and cytokine concentration was evaluated by Spearman’s rank correlation. For consistent direction, one up-regulated miRNA (miR-145) was found in four studies, and seven miRNAs were reported in three studies. One miRNA (miR-20a) and four miRNAs (miR-25-3p, miR-223, let-7f, and miR-20b) were reported in six and five studies. However, their expression was inconsistent. In the clinical validation, serum miR-145 was significantly down-regulated, whereas serum miR-20a was significantly up-regulated in NSCLC, compared with controls. Regarding serum cytokine, all cytokines [vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and transforming growth factor β (TGF-β)], except tumor necrosis factor-α (TNF-α), had a higher level in NSCLC patients than controls. In addition, we found a moderate correlation between the TGF-β concentration and miR-20a (r = −0.537, p = 0.002) and miR-223 (r = 0.428, p = 0.015) and a weak correlation between the VEGF concentration with miR-20a (r = 0.376, p = 0.037) and miR-223 (r = −0.355, p = 0.046). MiR-145 and miR-20a are potential biomarkers for NSCLC. In addition, the regulation of tumor-promoting cytokine, through miR-20a and miR-223, might be a new therapeutic approach for lung cancer.

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MicroRNA Isolation by Trizol-Based Method and Its Stability in Stored Serum and cDNA Derivatives

Cited by 23 publications

References 28 publications

MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer

MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer

Epigenetic Programming Through Breast Milk and Its Impact on Milk-Siblings Mating

Serum miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer

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