Selection and validation of endogenous controls for microRNA expression studies in endometrioid endometrial cancer tissues

Torres, Anna; Torres, Kamil; Wdowiak, Paulina; Paszkowski, Tomasz; Maciejewski, Ryszard

doi:10.1016/j.ygyno.2013.06.026

Cited by 68 publications

(61 citation statements)

References 40 publications

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“…For relative quantification of miRNA expression data were normalized using geometric mean of expression of experimentally chosen, stable endogenous controls RNU48, RNU44, U75 and U6 which were previously experimentally validated [12]. …”

Section: Methodsmentioning

confidence: 99%

In vitro and in vivo activity of miR-92a–Locked Nucleic Acid (LNA)–Inhibitor against endometrial cancer

et al. 2016

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BackgroundEndometrial cancer is the most common cancer of the female reproductive tract.Based on our previous studies we speculated that miR-92a exhibited pro-oncogenic properties in endometrial cancer, and therefore its inhibition could be used as a therapeutic measure in this disease. Therefore in the present study we aimed to investigate both in vitro and in vivo if inhibition of miR-92a in endometrial cancer would limit cancer cells proliferation.MethodsmiR-92a expression was evaluated in four endometrial cancer cell lines using qPCR. Inhibition of miR-92a activity was obtained in endometrial cancer cell lines by a transient transfection of a custom designed Locked Nucleic Acid (LNA)-Inhibitor, developed to work both in vitro and in vivo. In vitro proliferation studies were performed using xCELLigence RTCA DP system. In vivo experiment was performed in Cby.Cg-Foxn1 < nu>/cmdb mice bearing endometrial cancer xenografts, which were intraperitoneally injected with nine dosages of 25 mg/kg of miR-205-LNA-inhibitor.ResultsqPCR revealed increased expression of miR-92a in HEC-1-B, Ishikawa and AN3CA cells. LNA-i-miR-92a inhibited endometrial cancer growth in vitro. It was also demonstrated that systemic administration of LNA-i-miR-92a was feasible and exerted inhibitory effect on endometrial cancer xenograft growth in vivo with only mild toxic effects in treated animals, however the effect was observed until 12th experimental day and the last three dosages did not maintain the attenuating effect with the acceleration of tumor growth observed at the end and after cessation of the intraperitoneal therapy.ConclusionsTaken together, these results indicate that intraperitoneal delivery of miR-92a-LNA-modified-inhibitor is feasible, devoid of significant toxicity and moderately inhibits endometrial cancer growth in vivo, and therefore warrants further studies investigating other routes of inhibitor delivery possibly in other animal models.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2867-z) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

In vitro and in vivo activity of miR-92a–Locked Nucleic Acid (LNA)–Inhibitor against endometrial cancer

et al. 2016

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“…Two small nucleolar RNAs, RNU44 (ID 001094) and RNU48 (ID 001006) showing stable expression in human endometrium [21] were used as endogenous controls. cDNA synthesis was conducted from miRNA enriched RNA fraction with TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems, California, US).…”

Section: Methodsmentioning

confidence: 99%

High-Throughput Sequencing Approach Uncovers the miRNome of Peritoneal Endometriotic Lesions and Adjacent Healthy Tissues

et al. 2014

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Accumulating data have shown the involvement of microRNAs (miRNAs) in endometriosis pathogenesis. In this study, we used a novel approach to determine the endometriotic lesion-specific miRNAs by high-throughput small RNA sequencing of paired samples of peritoneal endometriotic lesions and matched healthy surrounding tissues together with eutopic endometria of the same patients. We found five miRNAs specific to epithelial cells – miR-34c, miR-449a, miR-200a, miR-200b and miR-141 showing significantly higher expression in peritoneal endometriotic lesions compared to healthy peritoneal tissues. We also determined the expression levels of miR-200 family target genes E-cadherin, ZEB1 and ZEB2 and found that the expression level of E-cadherin was significantly higher in endometriotic lesions compared to healthy tissues. Further evaluation verified that studied miRNAs could be used as diagnostic markers for confirming the presence of endometrial cells in endometriotic lesion biopsy samples. Furthermore, we demonstrated that the miRNA profile of peritoneal endometriotic lesion biopsies is largely masked by the surrounding peritoneal tissue, challenging the discovery of an accurate lesion-specific miRNA profile. Taken together, our findings indicate that only particular miRNAs with a significantly higher expression in endometriotic cells can be detected from lesion biopsies, and can serve as diagnostic markers for endometriosis.

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“…This finding indicates that some of these miRNAs could be, directly or indirectly, responsible for VEGF-A levels in endometrioid EC. A microarray study done by Torres et al [37] revealed regulation of several miRNAs in EEC tissues compared to normal endometrium. All miR-200 family members (miR-141, miR-200a, miR200b, miR-200c, and miR-429) were significantly upregulated, mostly in early stages of EEC.…”

Section: Endometrial Tumorsmentioning

confidence: 99%

“…Research into miRNAs thus carries great potential for a noninvasive diagnosis because it is, nowadays, possible to achieve miRNAs from the patients' plasma, and furthermore, aberrant expression can be also proven. For example, increased levels of miR-135b and miR-205 and decreased level of miR-30a-3p and miR-21 were identified in the plasma of EC patients, whereby after hysterectomy, levels of miR-135, miR-205, and miR-30a-3p decreased [39] respectively; signatures from EEC patients plasma-miR-9/miR-1228 and miR-9/miR92a-were also identified [37].…”

Section: Endometrial Tumorsmentioning

confidence: 99%

Malignant tumors of the uterine corpus: molecular background of their origin

et al. 2015

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Tumors of the uterine corpus can be divided into two main groups: endometrial tumors and mesenchymal tumors. The former ones are common gynecological diseases, whereas malignant mesenchymal tumors, which behave in a much more aggressive way, are quite rare with a poorer prognosis. The most common type of endometrial tumors is endometrioid adenocarcinomas, and in case of mesenchymal tumors, these are carcinosarcomas, or leiomyosarcomas, if only clear types of tumors are taken into account. The objective of this article is to review molecular-genetic abnormalities associated with tumorigenesis of both types of tumors, with focus on the most aggressive forms. This view includes a different expression pattern of genes, usually aberrant in cases of uterine cancer that can arise due to epigenetic modifications, mostly hypermethylation of promoters or microRNA (miRNA)'s interference with concrete genes. Furthermore, clinical predispositions of tumorigenesis, involving hormonal factors, age, and ethnicity, are also mentioned.

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Selection and validation of endogenous controls for microRNA expression studies in endometrioid endometrial cancer tissues

Cited by 68 publications

References 40 publications

In vitro and in vivo activity of miR-92a–Locked Nucleic Acid (LNA)–Inhibitor against endometrial cancer

In vitro and in vivo activity of miR-92a–Locked Nucleic Acid (LNA)–Inhibitor against endometrial cancer

High-Throughput Sequencing Approach Uncovers the miRNome of Peritoneal Endometriotic Lesions and Adjacent Healthy Tissues

Malignant tumors of the uterine corpus: molecular background of their origin

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