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Galectin-3-induced oxidized low-density lipoprotein promotes the phenotypic transformation of vascular smooth muscle cells.

Tian L, Chen K, Cao J, Han Z, Gao L, Wang Y, Fan Y, Wang C - Mol Med Rep (2015)

Bottom Line: The oxLDL‑induced cells exhibited increased expression levels of osteopontin, a smooth muscle synthetic protein, and calponin and α‑actin, smooth muscle contractile proteins.The oxLDL‑induced changes in cellular phenotype were associated with increased migration, proliferation, and phagocytosis.Concordant with these results, oxLDL‑treated smooth muscle cells exhibited activation of canonical Wnt signaling, as determined by an increase in the protein expression levels of β‑catenin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, P.R. China.

ABSTRACT
Oxidized low-density lipoprotein (oxLDL) is involved in the pathological phenotypic transformation of vascular smooth muscle cells in atherosclerosis. Galectin‑3 also has an important role in atherosclerosis. However, little is currently known regarding the effects of galectin‑3 on the oxLDL‑induced phenotypic transformation of vascular smooth muscle cells. In the present study, primary culture human umbilical vascular smooth muscle cells were treated with various oxLDL concentrations (0‑50 µg/ml) for 72 h, and phenotypic changes were subsequently recorded. The results of the present study suggested that oxLDL increases the expression levels of galectin‑3, and induces the phenotypic transformation of vascular smooth muscle cells. The oxLDL‑induced cells exhibited increased expression levels of osteopontin, a smooth muscle synthetic protein, and calponin and α‑actin, smooth muscle contractile proteins. The oxLDL‑induced changes in cellular phenotype were associated with increased migration, proliferation, and phagocytosis. Concordant with these results, oxLDL‑treated smooth muscle cells exhibited activation of canonical Wnt signaling, as determined by an increase in the protein expression levels of β‑catenin. Silencing of galectin‑3 by small interfering RNA reversed the phenotypic transformation and functional changes observed in the oxLDL‑treated cells, suggesting these changes were dependent on the activation of galectin‑3. In addition, galectin‑3 knockdown decreased the protein expression levels of β‑catenin in both the cytoplasm and nucleus; however, the mRNA expression levels of β‑catenin remained unchanged. These results suggest that galectin‑3 is responsible for the phenotypic transformation of human umbilical vascular smooth muscle cells, and the canonical Wnt/β-catenin signaling pathway may be involved in this process.

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Silencing of galectin-3 (gal-3) reversed the oxidized low-density lipoprotein (oxLDL)-induced phenotypic transformation of human umbilical smooth muscle cells (HUSMCs). HUSMCs were transfected with gal-3-specific small interfering (si)RNA for 24 h, and then cultured with 50 μg/ml oxLDL for 48 h. The mRNA and protein expression levels of gal-3, smooth muscle α-actin (SMA), calponin, and osteopontin (OPN) were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. (A and B) Western blotting and (C) RT-qPCR results of gal-3, calponin, OPN and SMA are shown. (B) The respective densitometric measurement results are given. The protein expression levels of gal-3, calponin, OPN and SMA were normalized to those of GAPDH. Band density of HUSMCs transfected with scramble siRNA was defined as control and set to 1. Data are presented as the mean ± standard deviation. *P<0.05, vs. the control. #P<0.05, vs. oxLDL.
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f2-mmr-12-04-4995: Silencing of galectin-3 (gal-3) reversed the oxidized low-density lipoprotein (oxLDL)-induced phenotypic transformation of human umbilical smooth muscle cells (HUSMCs). HUSMCs were transfected with gal-3-specific small interfering (si)RNA for 24 h, and then cultured with 50 μg/ml oxLDL for 48 h. The mRNA and protein expression levels of gal-3, smooth muscle α-actin (SMA), calponin, and osteopontin (OPN) were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. (A and B) Western blotting and (C) RT-qPCR results of gal-3, calponin, OPN and SMA are shown. (B) The respective densitometric measurement results are given. The protein expression levels of gal-3, calponin, OPN and SMA were normalized to those of GAPDH. Band density of HUSMCs transfected with scramble siRNA was defined as control and set to 1. Data are presented as the mean ± standard deviation. *P<0.05, vs. the control. #P<0.05, vs. oxLDL.

Mentions: To investigate the role of gal-3 in the oxLDL-induced phenotypic changes of HUSMCs, gal-3 knockdown was performed. Following transfection with siRNA, the expression of endogenous gal-3 was inhibited, and the phenotypic changes of HUSMCs were assessed. Gal-3-specific siRNA reduced the mRNA and protein expression levels of gal-3 by 95 and 45%, respectively. The non-targeting siRNA had no effect on gal-3 expression. Neither gal-3 nor the non-targeting siRNA induced a non-specific knockdown of GAPDH. Furthermore, gal-3 knockdown significantly inhibited both the oxLDL-induced mRNA and protein expression levels of OPN, calponin, and SMA (Fig. 2A-C). These results suggest that gal-3 may be involved in the oxLDL-induced phenotypic transformation of HUSMCs.


Galectin-3-induced oxidized low-density lipoprotein promotes the phenotypic transformation of vascular smooth muscle cells.

Tian L, Chen K, Cao J, Han Z, Gao L, Wang Y, Fan Y, Wang C - Mol Med Rep (2015)

Silencing of galectin-3 (gal-3) reversed the oxidized low-density lipoprotein (oxLDL)-induced phenotypic transformation of human umbilical smooth muscle cells (HUSMCs). HUSMCs were transfected with gal-3-specific small interfering (si)RNA for 24 h, and then cultured with 50 μg/ml oxLDL for 48 h. The mRNA and protein expression levels of gal-3, smooth muscle α-actin (SMA), calponin, and osteopontin (OPN) were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. (A and B) Western blotting and (C) RT-qPCR results of gal-3, calponin, OPN and SMA are shown. (B) The respective densitometric measurement results are given. The protein expression levels of gal-3, calponin, OPN and SMA were normalized to those of GAPDH. Band density of HUSMCs transfected with scramble siRNA was defined as control and set to 1. Data are presented as the mean ± standard deviation. *P<0.05, vs. the control. #P<0.05, vs. oxLDL.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581830&req=5

f2-mmr-12-04-4995: Silencing of galectin-3 (gal-3) reversed the oxidized low-density lipoprotein (oxLDL)-induced phenotypic transformation of human umbilical smooth muscle cells (HUSMCs). HUSMCs were transfected with gal-3-specific small interfering (si)RNA for 24 h, and then cultured with 50 μg/ml oxLDL for 48 h. The mRNA and protein expression levels of gal-3, smooth muscle α-actin (SMA), calponin, and osteopontin (OPN) were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. (A and B) Western blotting and (C) RT-qPCR results of gal-3, calponin, OPN and SMA are shown. (B) The respective densitometric measurement results are given. The protein expression levels of gal-3, calponin, OPN and SMA were normalized to those of GAPDH. Band density of HUSMCs transfected with scramble siRNA was defined as control and set to 1. Data are presented as the mean ± standard deviation. *P<0.05, vs. the control. #P<0.05, vs. oxLDL.
Mentions: To investigate the role of gal-3 in the oxLDL-induced phenotypic changes of HUSMCs, gal-3 knockdown was performed. Following transfection with siRNA, the expression of endogenous gal-3 was inhibited, and the phenotypic changes of HUSMCs were assessed. Gal-3-specific siRNA reduced the mRNA and protein expression levels of gal-3 by 95 and 45%, respectively. The non-targeting siRNA had no effect on gal-3 expression. Neither gal-3 nor the non-targeting siRNA induced a non-specific knockdown of GAPDH. Furthermore, gal-3 knockdown significantly inhibited both the oxLDL-induced mRNA and protein expression levels of OPN, calponin, and SMA (Fig. 2A-C). These results suggest that gal-3 may be involved in the oxLDL-induced phenotypic transformation of HUSMCs.

Bottom Line: The oxLDL‑induced cells exhibited increased expression levels of osteopontin, a smooth muscle synthetic protein, and calponin and α‑actin, smooth muscle contractile proteins.The oxLDL‑induced changes in cellular phenotype were associated with increased migration, proliferation, and phagocytosis.Concordant with these results, oxLDL‑treated smooth muscle cells exhibited activation of canonical Wnt signaling, as determined by an increase in the protein expression levels of β‑catenin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, P.R. China.

ABSTRACT
Oxidized low-density lipoprotein (oxLDL) is involved in the pathological phenotypic transformation of vascular smooth muscle cells in atherosclerosis. Galectin‑3 also has an important role in atherosclerosis. However, little is currently known regarding the effects of galectin‑3 on the oxLDL‑induced phenotypic transformation of vascular smooth muscle cells. In the present study, primary culture human umbilical vascular smooth muscle cells were treated with various oxLDL concentrations (0‑50 µg/ml) for 72 h, and phenotypic changes were subsequently recorded. The results of the present study suggested that oxLDL increases the expression levels of galectin‑3, and induces the phenotypic transformation of vascular smooth muscle cells. The oxLDL‑induced cells exhibited increased expression levels of osteopontin, a smooth muscle synthetic protein, and calponin and α‑actin, smooth muscle contractile proteins. The oxLDL‑induced changes in cellular phenotype were associated with increased migration, proliferation, and phagocytosis. Concordant with these results, oxLDL‑treated smooth muscle cells exhibited activation of canonical Wnt signaling, as determined by an increase in the protein expression levels of β‑catenin. Silencing of galectin‑3 by small interfering RNA reversed the phenotypic transformation and functional changes observed in the oxLDL‑treated cells, suggesting these changes were dependent on the activation of galectin‑3. In addition, galectin‑3 knockdown decreased the protein expression levels of β‑catenin in both the cytoplasm and nucleus; however, the mRNA expression levels of β‑catenin remained unchanged. These results suggest that galectin‑3 is responsible for the phenotypic transformation of human umbilical vascular smooth muscle cells, and the canonical Wnt/β-catenin signaling pathway may be involved in this process.

Show MeSH
Related in: MedlinePlus