2009
DOI: 10.2119/molmed.2009.00030
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Role of E2F1-Cyclin E1-Cyclin E2 Circuit in Human Coronary Smooth Muscle Cell Proliferation and Therapeutic Potential of Its Downregulation by siRNAs

Abstract: Aberrant coronary vascular smooth muscle cell (CSMC) proliferation is a pivotal event underlying intimal hyperplasia, a phenomenon impairing the long-term efficacy of bypass surgery and angioplasty procedures. Consequently research has become focused on efforts to identify molecules that are able to control CSMC proliferation. We investigated downregulation of CSMC growth by small interfering RNAs (siRNAs) targeted against E2F1, cyclin E1, and cyclin E2 genes, whose contribution to CSMC proliferation is only n… Show more

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Cited by 43 publications
(33 citation statements)
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“…We typically perform the selection by testing different amounts of a given lipid, different ratio lipid/siRNA, different transfection times and different transfection medium [103][104][105]. In parallel to the above mentioned tests, we also evaluate the transfection efficiency and siRNA cellular distribution by using FITC-labelled siRNAs.…”
Section: Genuine Sirna Off-targetingmentioning
confidence: 99%
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“…We typically perform the selection by testing different amounts of a given lipid, different ratio lipid/siRNA, different transfection times and different transfection medium [103][104][105]. In parallel to the above mentioned tests, we also evaluate the transfection efficiency and siRNA cellular distribution by using FITC-labelled siRNAs.…”
Section: Genuine Sirna Off-targetingmentioning
confidence: 99%
“…In contrast, a more contained proteomic analysis limited to some proteins interacting with the target protein, are technically easier to perform, still being reasonably informative. For example, while studying the effects of the depletion of the transcription factor E2F1 [104], we evaluated also the effects on the protein levels of some of its transcriptional regulated proteins such as cyclin E1 and E2. A higher, although still limited, amount of protein interactions can be evaluated by a recently developed approach.…”
Section: Experimental Approaches To Detect Sirna Off-targeting and Spmentioning
confidence: 99%
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“…The sequences of the siRNAs directed against the transcription factor E2F1 mRNA (siE2F1) and the luciferase mRNA (siGL2, control), were purchased from Eurofins Genomics GmBH, Ebersberg, D. The sequences of these chemically un-modified siRNAs, have been previously reported Dapas et al, 2009). The day before transfection, JHH6 cells were seeded at a density of 3.8 Á 10 3 cells/cm 2 in 6 well microplate in the presence of 3 ml of 10% fetal calf serum-containing medium.…”
Section: In Vitro Transfection and Determination Of Target Mrna/protementioning
confidence: 99%
“…Compared to the conventional therapeutic drugs, siRNAs represent an emerging paradigm for the treatment of many human diseases such as cancer Resnier et al, 2013), cardiovascular, neurodegenerative, metabolic diseases and viral illnesses (Werth et al, 2010;Dapas et al, 2009;Racchi et al, 2009;Deng et al, 2014). However, owing to their negative charges, siRNA molecules are not readily taken up by cells that have a negatively charged surface.…”
Section: Introductionmentioning
confidence: 99%