Identification of ETFB as a candidate protein that participates in the mechanoregulation of fibroblast cell number in collagen gel culture

Hirokawa, Shigenari; Shimanuki, Tomomasa; Kohno, Hiroyuki; Nishimori, Yasutomo; Shimosaka, Makoto

doi:10.1016/j.jdermsci.2011.08.003

Cited by 6 publications

(13 citation statements)

References 39 publications

(63 reference statements)

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“…The electron transfer flavoprotein b subunit (ETFB) was one of the candidates identified [1]. In the present report, we verified the inhibitory effect of ETFB on fibroblast cell number in our 3D collagen culture system and determined the ETFB mRNA expression pattern.…”

supporting

confidence: 64%

“…We previously reported that knockdown of ETFB inhibited mechanoregulated cell number in fibroblast-populated 3D collagen gel cultures [1]. To confirm these findings, we transfected fibroblasts with ETFB or negative control siRNA and then counted cell numbers every day for 4 days of attached or detached collagen gel culture (Fig.…”

Section: Resultsmentioning

confidence: 55%

“…ETFB knockdown was initially shown to reduce mechanoregulated cell number [1]. Because ETFB is involved in electron transfer during the b-oxidation cascade [24,25], it is possible that the inhibitory effect of ETFB knockdown affects multiple cellular processes in treated fibroblasts.…”

Section: Resultsmentioning

confidence: 99%

“…We identified ETFB in a previous loss-of-function screen for factors that affect mechanoregulated cell proliferation in collagen gel culture [1]. As we previously described [1], it has been reported that the 3D collagen culture system can mimic the proliferative or quiescent state of fibroblasts by changing mechanical stress by tethering the well (attached culture) or floating freely in the medium (detached culture).…”

Section: Discussionmentioning

confidence: 99%

“…The collagen gel culture was performed as previously described [1]. Briefly, for one 24-well plate, a collagen matrix solution was prepared by mixing 7 mL of 5 mg/mL bovine dermis-derived, acidsolubilized type I collagen (Cellgen, Koken, Tokyo), 3.5 mL FBS, 7 mL of 5Â DMEM (4500 mg/mL glucose, final concentration, HEPES(À), NaHCO 3 (À); 12800-017, Invitrogen, CA), 3.5 mL of 200 mM HEPES, 3.5 mL of 2.2% NaHCO 3 , 1.75 mL of 0.1 N NaOH and 5.25 mL of H 2 O.…”

Section: Cell Culture and Reagentsmentioning

confidence: 99%

See 4 more Smart Citations

Knockdown of electron transfer flavoprotein β subunit reduced TGF-β-induced α-SMA mRNA expression but not COL1A1 in fibroblast-populated three-dimensional collagen gel cultures

Hirokawa

Shimanuki

Kohno

et al. 2012

Journal of Dermatological Science

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supporting

confidence: 64%

Section: Resultsmentioning

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Cell Culture and Reagentsmentioning

confidence: 99%

See 3 more Smart Citations

Knockdown of electron transfer flavoprotein β subunit reduced TGF-β-induced α-SMA mRNA expression but not COL1A1 in fibroblast-populated three-dimensional collagen gel cultures

Hirokawa

Shimanuki

Kohno

et al. 2012

Journal of Dermatological Science

Self Cite

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Phosphoproteomic response of cardiac endothelial cells to ischemia and ultrasound

Emechebe

Giraud

Ammi

et al. 2021

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Identification of potential biomarkers for predicting the early onset of diabetic cardiomyopathy in a mouse model

Johnson

Nxele

Cour

et al. 2020

Sci Rep

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Type 2 diabetes (T2D) is characterized by metabolic derangements that cause a shift in substrate preference, inducing cardiac interstitial fibrosis. Interstitial fibrosis plays a key role in aggravating left ventricular diastolic dysfunction (LVDD), which has previously been associated with the asymptomatic onset of heart failure. The latter is responsible for 80% of deaths among diabetic patients and has been termed diabetic cardiomyopathy (DCM). Through in silico prediction and subsequent detection in a leptin receptor-deficient db/db mice model (db/db), we confirmed the presence of previously identified potential biomarkers to detect the early onset of DCM. Differential expression of Lysyl Oxidase Like 2 (LOXL2) and Electron Transfer Flavoprotein Beta Subunit (ETFβ), in both serum and heart tissue of 6-16-week-old db/db mice, correlated with a reduced left-ventricular diastolic dysfunction as assessed by high-resolution Doppler echocardiography. Principal component analysis of the combined biomarkers, LOXL2 and ETFβ, further displayed a significant difference between wild type and db/ db mice from as early as 9 weeks of age. Knockdown in H9c2 cells, utilising siRNA of either LOXL2 or ETFβ, revealed a decrease in the expression of Collagen Type I Alpha1 (COL1A1), a marker known to contribute to enhanced myocardial fibrosis. Additionally, receiver-operating curve (ROC) analysis of the proposed diagnostic profile showed that the combination of LOXL2 and ETFβ resulted in an area under the curve (AUC) of 0.813, with a cutoff point of 0.824, thus suggesting the favorable positive predictive power of the model and further supporting the use of LOXL2 and ETFβ as possible early predictive DCM biomarkers. Diabetes affects 463 million people worldwide and this number is said to increase to 700 million by 2045 1. Diabetes and its complications are inextricably linked to cardiovascular dysfunction, which is currently the leading cause of mortality worldwide, affecting 17.9 million individuals 2. This association was first reported in the Framingham Heart study and since then numerous reports have come to the forefront, confirming a 2-4 times increased susceptibility of diabetic individuals to heart failure (HF) 3,4. Coronary artery disease (CAD) is the major type of CVD responsible for HF in diabetic individuals, however diabetic cardiomyopathy (DCM) is an established complication of diabetes mellitus (DM) existing in the absence of CAD or hypertension 5. Furthermore, DCM is referred to as the silent killer, due to the manifestation of a long subclinical period in which the disease exists with no overt clinical symptoms 6-8 .

show abstract

Identification of ETFB as a candidate protein that participates in the mechanoregulation of fibroblast cell number in collagen gel culture

Cited by 6 publications

References 39 publications

Knockdown of electron transfer flavoprotein β subunit reduced TGF-β-induced α-SMA mRNA expression but not COL1A1 in fibroblast-populated three-dimensional collagen gel cultures

Knockdown of electron transfer flavoprotein β subunit reduced TGF-β-induced α-SMA mRNA expression but not COL1A1 in fibroblast-populated three-dimensional collagen gel cultures

Phosphoproteomic response of cardiac endothelial cells to ischemia and ultrasound

Identification of potential biomarkers for predicting the early onset of diabetic cardiomyopathy in a mouse model

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