Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes

Jørgensen, Stine; Baker, Adam; Möller, Sören; Nielsen, Boye Schnack

doi:10.1016/j.ymeth.2010.07.002

Cited by 168 publications

(129 citation statements)

References 27 publications

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“…A similar scenario may contribute to breast cancer development : miR-9, miR-19b, miR-146, miR-181c, miR-183, miR-200c, miR-205, miR-223, miR-423, miR-425 Y: let-7a, miR-32, miR-33b, miR-369, miR-409, miR-424, miR-431, miR-451, miR-496, miR-503 miR-516 Ratner et al (2010) EEC carcinoma versus benign tissue from control patients [: miR-34a, miR-182, miR-183, miR-200a, miR-205, miR-572, miR-622, miR-650 Y: miR-411, miR-487b Chung et al (2009) EEC versus benign tissue from control patients [: miR-10a, miR-23a, miR-25, miR-28, miR-34a, miR-95, miR-103, miR-106a, miR-107, miR-130b, miR-141, miR-151, miR-155, miR-17-5p, miR-182, miR-183, miR-184, miR-191, miR-194, miR-200a, miR-200c, miR-203, miR-205, miR-210 : miR-10a, miR-31, miR-96, miR-133b, miR-141, miR-142-5p, miR-155, miR-182, miR-200a, miR-200b, miR-200c, miR-203, miR-205, miR-210, miR-363, miR-429, miR-432, miR-449 Y: miR-99b, miR-133, miR-193a, miR-193b, miR-204, miR-368 Boren et al (2008) EEC versus atypical hyperplasia versus normal endometrium [: let-7c, miR-103, miR-106a, miR-107, miR-181a, miR-185, miR-210, miR-423 Y: let-7i, miR-30c, miR-152, miR-193, miR-221 MiRNA represents a new class of biomarkers that can complement existing conventional markers, including metabolites, antigens and mRNA transcripts. The fact that miRNAs remain largely intact in formalin-fixed, paraffinembedded clinical samples highlights their potential in molecular phenotyping of benign and malignant reproductive disorders as well as for monitoring treatment efficacy (Bitossi et al 2008, Szafranska et al 2008, Tam 2008, Jorgensen et al 2010, Nuovo 2010, Schuster et al 2010. Furthermore, recent evidence suggests that some neoplastic processes, for example, ovarian cancer, generate mRNA profiles in blood cells characteristic of the tumours, suggesting that whole blood miRNA profiling could be used for diagnostic purposes (Balch et al 2009, Lodes et al 2009, Resnick et al 2009, Hausler et al 2010.…”

Section: The Mirna-foxo Axis In Endometrial Carcinogenesismentioning

confidence: 99%

The diversity of sex steroid action: the role of micro-RNAs and FOXO transcription factors in cycling endometrium and cancer

Lam¹,

Shah²,

Brosens³

2011

Journal of Endocrinology

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The rise and fall in ovarian oestrogen and progesterone production orchestrates a series of events that are indispensable for reproduction, including ovulation, implantation, decidualisation and menstruation. In the uterus, these events involve extensive tissue remodelling, characterised by waves of endometrial cell proliferation, differentiation, recruitment of inflammatory cells, apoptosis, tissue breakdown, menstruation and regeneration. The ability of ovarian hormones to trigger such diverse physiological responses is foremost dependent upon interaction of activated steroid receptors with specific transcription factors, such as Forkhead box class O (FOXO) proteins, involved in cell fate decisions. Furthermore, micro-RNAs (miRNAs), small non-coding RNAs that function as posttranscriptional regulators of gene expression, have emerged as a major regulator system of steroid hormone responses in the female reproductive tract. Consequently, increasing evidence shows that deregulated uterine miRNA expression underpins a spectrum of common reproductive disorders, ranging from implantation failure to endometriosis. Furthermore, by targeting FOXO transcription factors and other key regulators of tissue homeostasis, oncogenic endometrial miRNAs promote tumourigenesis and cancer progression.

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Section: The Mirna-foxo Axis In Endometrial Carcinogenesismentioning

confidence: 99%

The diversity of sex steroid action: the role of micro-RNAs and FOXO transcription factors in cycling endometrium and cancer

Lam¹,

Shah²,

Brosens³

2011

Journal of Endocrinology

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“…ISH was performed as described elsewhere (21). In brief, ISH was carried out on 6-µm tissue sections containing CRC and normal colorectal tissue using double DIG-labelled LNA probes for human miRNA-126 (Exiqon A/S, Vedbaek, Denmark).…”

Section: Cd105 and Caldesmon Immunostaining And Mvd Countingmentioning

confidence: 99%

Elevated microRNA-126 is associated with high vascular endothelial growth factor receptor 2 expression levels and high microvessel density in colorectal cancer

Hansen

Andersen

Nielsen³

et al. 2011

Oncology Letters

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Abstract. MicroRNAs (miRNAs) are involved in a number of biological processes, including tumour biology. Pre-clinical studies have shown that miRNA-126 regulates signalling downstream of vascular endothelial growth factor receptor 2 (VEGFR-2) and, consequently, angiogenesis. The aim of this study was to analyse the possible relationship between miRNA-126, VEGFR-2 and angiogenesis in tumour tissue from patients with colorectal cancer (CRC). Tumour tissue was obtained from 81 patients. The miRNA-126 and VEGFR-2 gene expression levels were analysed by PCR and the protein concentrations of VEGFR-2 were analysed by ELISA. Angiogenesis, visualised by the endothelial cell marker CD105 combined with caldesmon, was assessed by immunohistochemistry and the microvessel density (MVD) technique. In situ hybridisation was performed for miRNA-126. Tumours were classified as low or high miRNA-126-expressing using the median as the cut-off. The median gene expression levels of VEGFR-2 were significantly lower in the tumours expressing low levels of miRNA-126, 0.30 (95% CI, 0.24-0.36), compared to those expressing high levels of miRNA-126, 0.48 (95% CI, 0.28-0.60), p=0.02. A positive association was observed with VEGFR-2 protein concentrations, p=0.06. The median MVD was significantly lower in the tumours expressing low levels of 5.8 (95% CI,), compared to those expressing high levels, 8.0 (95% CI, 6.33-9.00), p<0.01. miRNA-126 was detected in endothelial cells by in situ hybridisation analysis. These results suggest that high levels of miRNA-126 in CRC are associated with high VEGFR-2 mRNA and protein levels and a higher density of newly formed microvessels. However, further studies should be conducted to analyse the clinical value of miRNA-126 in CRC.

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“…The extent of proteinase-K digestion (which depends on the concentration, incubation temperature and time) and the hybridization temperature which has been found to provide a fairly high signal-to-noise ratio at 30° C below the measured Tm [74]. As for other methodologies, negative and positive controls are pivotal.…”

Section: Analysis Performed Atmentioning

confidence: 99%

miRNA Detection Methods and Clinical Implications in Lung Cancer

et al. 2014

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Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes

Cited by 168 publications

References 27 publications

The diversity of sex steroid action: the role of micro-RNAs and FOXO transcription factors in cycling endometrium and cancer

The diversity of sex steroid action: the role of micro-RNAs and FOXO transcription factors in cycling endometrium and cancer

Elevated microRNA-126 is associated with high vascular endothelial growth factor receptor 2 expression levels and high microvessel density in colorectal cancer

miRNA Detection Methods and Clinical Implications in Lung Cancer

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