MicroRNA miR-145 inhibits proliferation, invasiveness, and stem cell phenotype of an in vitro endometriosis model by targeting multiple cytoskeletal elements and pluripotency factors

Adammek, M; Greve, Burkhard; Kässens, Nadja; Schneider, Cornelia; Brüggemann, Kathrin; Schüring, Andreas N.; Starzinski‐Powitz, Anna; Kiesel, L.; Götte, Martin

doi:10.1016/j.fertnstert.2012.11.055

Cited by 86 publications

(100 citation statements)

References 39 publications

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“…It has been reported that miR-145 is expressed at low levels in breast cancer, colon cancer, lung cancer and other tumor tissues compared with normal tissues (14). As previously demonstrated, by transfecting synthetic miR-145 oligonucleotides into neuroblastoma cells, endometrial stem cells, and bladder cancer cells, the expression of genes, such as insulin-like growth factor receptor I (IGF-IR) and hypoxia-inducible factor 2α (HIF-2α) was downregulated, which resulted in the inhibition of cancer cell growth and invasion (15)(16)(17).…”

Section: Introductionmentioning

confidence: 94%

“…miR-145 was reverse transcribed into cDNA using an miRcute miRNA cDNA kit and miRcute miRNA qPCR detection kits, to measure the expression levels of miR-145. The PCR cycles were as follows: denaturation at 94˚C for 2 min, followed by 40 cycles at 94˚C for 20 sec and 60˚C for 34 sec and snRU6 was used as an internal normalization control as previously described (16). The relative changes in miRNA expression were calculated using the 2 -ΔΔCT method.…”

Section: Analysis Of Endogenous Mir-145 Expression Levels In Differenmentioning

confidence: 99%

“…Cell proliferation was measured using by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as previously described (16). Following transfection for 48 h, 10,000 HepG2 cells were seeded in 96-well plates and cultured for 24 h. Briefly, MTT solution was added and followed by further incubation for 4 h; DMSO was then added to dissolve the formazan crystals and the optical density was measured at 570 and 630 nm using a microplate reader (Multiskan MK3, Thermo Fisher Scientific Inc.,Waltham, MA, USA).…”

Section: Analysis Of Endogenous Mir-145 Expression Levels In Differenmentioning

confidence: 99%

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Optimization of a cationic liposome-based gene delivery system for the application of miR-145 in anticancer therapeutics

Jin

Ding

Che

et al. 2016

International Journal of Molecular Medicine

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Abstract. In order to improve the delivery efficiency of microRNA (miRNA or miR)-145, the present study examined several factors which may affect cationic liposome (CL)-based transfection, including the hydration medium used for the preparation of liposomes, the quantity of the plasmid, the molar ratio of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP)/cholesterol (chol), or DOTAP/chol, and the weight ratio of DOTAP/DNA. In order to enhance the transfection efficiency, protamine was selected as a DNA-condensing agent to form liposome-protamine-DNA (LPD) ternary complexes. An agarose gel retardation assay was used to examine the DNA binding affinity of the CLs. Following transfection, GFP fluorescence images were captured and flow cytometry was performed to determine the transfection efficiency. Furthermore, an MTT assay was performed to determine the cytotoxicity of the liposome complexes. The final optimal conditions were as follows: 5% glucose as the hydration medium, a molar ratio of DOTAP/chol at 3:1 for the preparation of CLs, a weight ratio of DOTAP/protamine/DNA of 3:0.5:1, with 8 µg plasmid added for the preparation of the LPD complexes. In vitro, the LPD complexes exhibited an enhanced transfection efficiency and low cytotoxicity, which indicated that the presented LPD vector enhanced the transfection efficiency of the CLs. The HepG2 cells were found to have the lowest expression levels of miR-145out of the cell lines tested (A549, BGC-823, HepG2, HeLa, LoVo and MCF-7). Following the transient transfection of the HepG2 cells with miR-145, the results revealed that the overexpression of miR-145 inhibited the proliferation of the HepG2 cells and downregulated the expression of cyclin-dependent kinase 6 (CDK6), cyclinD1, c-Myc, and Sp1 transcription factor (Sp1). In conclusion, in this study, we optimized a liposome-based delivery system for the efficient delivery of miR-145 into cancer cells. This may provide a foundation for further research into the use of miR-145 in anticancer therapeutics.

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Section: Introductionmentioning

confidence: 94%

Section: Analysis Of Endogenous Mir-145 Expression Levels In Differenmentioning

confidence: 99%

Section: Analysis Of Endogenous Mir-145 Expression Levels In Differenmentioning

confidence: 99%

See 1 more Smart Citation

Optimization of a cationic liposome-based gene delivery system for the application of miR-145 in anticancer therapeutics

Jin

Ding

Che

et al. 2016

International Journal of Molecular Medicine

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“…miR9 and miR-374 directly target the 3′-untranslated region of claudin14 mRNA, leading to claudin14 mRNA translational repression and decay, in a cooperative manner [28] . miR145 impairs TJ function by repressing junctional adhesion molecule1 expression [29] .…”

Section: Mirnas and Intestinal Tissue Barriersmentioning

confidence: 99%

miRNAs as new molecular insights into inflammatory bowel disease: Crucial regulators in autoimmunity and inflammation

Zhang

2016

WJG

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Inflammatory bowel disease (IBD) is characterized by chronic relapsing inflammatory disorders of the gastrointestinal tract, and includes two major phenotypes: ulcerative colitis and Crohn's disease. The pathogenesis of IBD is not fully understood as of yet. It is believed that IBD results from complicated interactions between environmental factors, genetic predisposition, and immune disorders. miRNAs are a class of small non-coding RNAs that can regulate gene expression by targeting the 3′-untranslated region of specific mRNAs for degradation or translational inhibition. miRNAs are considered to play crucial regulatory roles in many biologic processes, such as immune cellular differentiation, proliferation, and apoptosis, and maintenance of immune homeostasis. Recently, aberrant expression of miRNAs was revealed to play an important role in autoimmune diseases, including IBD. In this review, we discuss the current understanding of how miRNAs regulate autoimmunity and inflammation by affecting the differentiation, maturation, and function of various immune cells. In particular, we focus on describing specific miRNA expression profiles in tissues and peripheral blood that may be associated with the pathogenesis of IBD. In addition, we summarize the opportunities for utilizing miRNAs as new biomarkers and as potential therapeutic targets in IBD.

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“…For example, miRNA-145 and OCT4, one of the key transcription factors required to maintain the pluripotent state of embryonic stem cells, suppress each other's expression ( Fig. 1; Xu et al 2009;Wu et al 2011;Adammek et al 2013). In embryonic stem cells, high levels of OCT4 lead to repression of miR-145 expression, while a reversed pattern is present in differentiated cells.…”

mentioning

confidence: 99%

Small but sturdy: small RNAs in cellular memory and epigenetics

2014

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Cells in multicellular organisms have distinct identities characterized by their profiles of expressed genes. Cell identities can be stable over a long time and through multiple cellular divisions but are also responsive to extracellular signals. Since the DNA sequence is identical in all cells, a “cellular memory” of expression profiles is achieved by what are defined as epigenetic mechanisms. Two major molecular principles—networks of transcription factors and maintenance of cis-chromatin modifications—have been implicated in maintaining cellular memory. Here we describe recent studies demonstrating that short noncoding RNAs can also provide molecular signals that define epigenetic states of cells. Small RNAs can act independently or cooperate with chromatin modifications to achieve long-lasting effects necessary for cellular memory and transgenerational inheritance.

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MicroRNA miR-145 inhibits proliferation, invasiveness, and stem cell phenotype of an in vitro endometriosis model by targeting multiple cytoskeletal elements and pluripotency factors

Cited by 86 publications

References 39 publications

Optimization of a cationic liposome-based gene delivery system for the application of miR-145 in anticancer therapeutics

Optimization of a cationic liposome-based gene delivery system for the application of miR-145 in anticancer therapeutics

miRNAs as new molecular insights into inflammatory bowel disease: Crucial regulators in autoimmunity and inflammation

Small but sturdy: small RNAs in cellular memory and epigenetics

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