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Up-regulation of stomatin expression by hypoxia and glucocorticoid stabilizes membrane-associated actin in alveolar epithelial cells.

Chen JC, Cai HY, Wang Y, Ma YY, Song LN, Yin LJ, Cao DM, Diao F, Li YD, Lu J - J. Cell. Mol. Med. (2013)

Bottom Line: Stomatin is an important lipid raft-associated protein which interacts with membrane proteins and plays a role in the membrane organization.In this study we found that hypoxia and dexamethasone (dex), a synthetic GC not only up-regulated the expression of stomatin alone, but also imposed additive effect on the expression of stomatin in A549 cells, primary AEC and lung of rats.Moreover, we found that hypoxia exposure did not affect membrane-associated actin, but decreased actin in cytoplasm in A549 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, The Second Military Medical University, Shanghai, China.

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Hypoxia and dex had additive effects on up-regulation of stomatin expression in vitro and in vivo. A549 cells were exposed to hypoxia (1% O2, 5% CO2, 94% N2), or treated with 10 nM dex, or co-treated with hypoxia and dex for 16 hrs. Control cells were cultured in a normoxia incubator (with 5% CO2, 20% O2). The levels of stomatin mRNA and protein were assessed by real-time PCR (A) and Western blot (B). (C) Primary alveolar epithelial cells (AEC) were isolated and cultured to fourth or fifth passage. About 4 × 105 cells/dish were treated as described above and stomatin mRNA were assessed. β-actin was used as a normalization control for real-time PCR and Western blot. Data were represented as mean ± S.D. of three independent experiments.*P < 0.05;**P < 0.01; #P < 0.01 versus Dex. Adult male rats with adx were exposed to hypoxia (8% O2 and 92% N2), or treated with dex (5 mg/kg), or co-treated with both hypoxia and dex for 12 hrs. Sham rats were treated with or without hypoxia (n = 6–9). The levels of stomatin mRNA and protein were assessed by real-time PCR (D) and Western blot (E). Stomatin level was normalized to that of β-actin. Values were expressed as mean ± S.D. *P < 0.05 versus sham, #P < 0.05 versus adx.
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fig03: Hypoxia and dex had additive effects on up-regulation of stomatin expression in vitro and in vivo. A549 cells were exposed to hypoxia (1% O2, 5% CO2, 94% N2), or treated with 10 nM dex, or co-treated with hypoxia and dex for 16 hrs. Control cells were cultured in a normoxia incubator (with 5% CO2, 20% O2). The levels of stomatin mRNA and protein were assessed by real-time PCR (A) and Western blot (B). (C) Primary alveolar epithelial cells (AEC) were isolated and cultured to fourth or fifth passage. About 4 × 105 cells/dish were treated as described above and stomatin mRNA were assessed. β-actin was used as a normalization control for real-time PCR and Western blot. Data were represented as mean ± S.D. of three independent experiments.*P < 0.05;**P < 0.01; #P < 0.01 versus Dex. Adult male rats with adx were exposed to hypoxia (8% O2 and 92% N2), or treated with dex (5 mg/kg), or co-treated with both hypoxia and dex for 12 hrs. Sham rats were treated with or without hypoxia (n = 6–9). The levels of stomatin mRNA and protein were assessed by real-time PCR (D) and Western blot (E). Stomatin level was normalized to that of β-actin. Values were expressed as mean ± S.D. *P < 0.05 versus sham, #P < 0.05 versus adx.

Mentions: The effect of dex on stomatin expression in A549 cells was investigated using RT PCR and Western blot. Cells were treated with different concentrations of dex (0.1–100 nM) for 12 hrs, or 10 nM dex for different times. As shown in Figure 2A and B, dex increased the mRNA level of stomatin in a concentration- and time-dependent manner. The level of stomatin mRNA in A549 cells treated with 10 nM dex for 8 hrs was about 3.8-fold of that in control cells (P < 0.01). Ten nanomole dex also significantly increased stomatin expression at protein level in a time-dependent fashion (Fig. 2C). The stomatin mRNA induced by dex was significantly blocked by RU486, an antagonist of GR (Fig. 3D), indicating that the effects of dex were mediated through GR.


Up-regulation of stomatin expression by hypoxia and glucocorticoid stabilizes membrane-associated actin in alveolar epithelial cells.

Chen JC, Cai HY, Wang Y, Ma YY, Song LN, Yin LJ, Cao DM, Diao F, Li YD, Lu J - J. Cell. Mol. Med. (2013)

Hypoxia and dex had additive effects on up-regulation of stomatin expression in vitro and in vivo. A549 cells were exposed to hypoxia (1% O2, 5% CO2, 94% N2), or treated with 10 nM dex, or co-treated with hypoxia and dex for 16 hrs. Control cells were cultured in a normoxia incubator (with 5% CO2, 20% O2). The levels of stomatin mRNA and protein were assessed by real-time PCR (A) and Western blot (B). (C) Primary alveolar epithelial cells (AEC) were isolated and cultured to fourth or fifth passage. About 4 × 105 cells/dish were treated as described above and stomatin mRNA were assessed. β-actin was used as a normalization control for real-time PCR and Western blot. Data were represented as mean ± S.D. of three independent experiments.*P < 0.05;**P < 0.01; #P < 0.01 versus Dex. Adult male rats with adx were exposed to hypoxia (8% O2 and 92% N2), or treated with dex (5 mg/kg), or co-treated with both hypoxia and dex for 12 hrs. Sham rats were treated with or without hypoxia (n = 6–9). The levels of stomatin mRNA and protein were assessed by real-time PCR (D) and Western blot (E). Stomatin level was normalized to that of β-actin. Values were expressed as mean ± S.D. *P < 0.05 versus sham, #P < 0.05 versus adx.
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fig03: Hypoxia and dex had additive effects on up-regulation of stomatin expression in vitro and in vivo. A549 cells were exposed to hypoxia (1% O2, 5% CO2, 94% N2), or treated with 10 nM dex, or co-treated with hypoxia and dex for 16 hrs. Control cells were cultured in a normoxia incubator (with 5% CO2, 20% O2). The levels of stomatin mRNA and protein were assessed by real-time PCR (A) and Western blot (B). (C) Primary alveolar epithelial cells (AEC) were isolated and cultured to fourth or fifth passage. About 4 × 105 cells/dish were treated as described above and stomatin mRNA were assessed. β-actin was used as a normalization control for real-time PCR and Western blot. Data were represented as mean ± S.D. of three independent experiments.*P < 0.05;**P < 0.01; #P < 0.01 versus Dex. Adult male rats with adx were exposed to hypoxia (8% O2 and 92% N2), or treated with dex (5 mg/kg), or co-treated with both hypoxia and dex for 12 hrs. Sham rats were treated with or without hypoxia (n = 6–9). The levels of stomatin mRNA and protein were assessed by real-time PCR (D) and Western blot (E). Stomatin level was normalized to that of β-actin. Values were expressed as mean ± S.D. *P < 0.05 versus sham, #P < 0.05 versus adx.
Mentions: The effect of dex on stomatin expression in A549 cells was investigated using RT PCR and Western blot. Cells were treated with different concentrations of dex (0.1–100 nM) for 12 hrs, or 10 nM dex for different times. As shown in Figure 2A and B, dex increased the mRNA level of stomatin in a concentration- and time-dependent manner. The level of stomatin mRNA in A549 cells treated with 10 nM dex for 8 hrs was about 3.8-fold of that in control cells (P < 0.01). Ten nanomole dex also significantly increased stomatin expression at protein level in a time-dependent fashion (Fig. 2C). The stomatin mRNA induced by dex was significantly blocked by RU486, an antagonist of GR (Fig. 3D), indicating that the effects of dex were mediated through GR.

Bottom Line: Stomatin is an important lipid raft-associated protein which interacts with membrane proteins and plays a role in the membrane organization.In this study we found that hypoxia and dexamethasone (dex), a synthetic GC not only up-regulated the expression of stomatin alone, but also imposed additive effect on the expression of stomatin in A549 cells, primary AEC and lung of rats.Moreover, we found that hypoxia exposure did not affect membrane-associated actin, but decreased actin in cytoplasm in A549 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, The Second Military Medical University, Shanghai, China.

Show MeSH
Related in: MedlinePlus