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Tuberin-deficiency downregulates N-cadherin and upregulates vimentin in kidney tumor of TSC patients.

Liang S, Salas T, Gencaslan E, Li B, Habib SL - Oncotarget (2014)

Bottom Line: In addition, cells treated with rapamycin showed a significant increase in p-Akt and a decrease in p-p70S6K that was associated with a decrease expression of vimentin and a slight increase expression in N-cadherin.On the other hand, cells treated with Akt inhibitor revealed a significant decrease in p-Akt and p-p70S6K that was associated with a significant decrease in vimentin and an increase in N-cadherin expression.Kidney tumors from TSC patients showed significant decrease in N-cadherin and significant increased in vimentin protein expression compared to control kidney tissues.

View Article: PubMed Central - PubMed

Affiliation: South Texas Veterans Health Care System, San Antonio, TX.

ABSTRACT
Angiomyolipomas (AMLs) are associated with cell fibrosis in kidney of Tuberous Sclerosis Complex patients. The mechanism by which the fibrotic proteins accumulated in AMLs has not been explored. In the present study, we investigated the role of Akt/tuberin/mTOR pathway in the regulation cell fibrosis proteins. AML cells that expressed low levels of tuberin showed less expression of N-cadherin and higher of vimentin proteins compared to HEK293 cells. AML cells infected with Ad-tuberin showed a significant decrease in vimentin and an increase in N-cadherin protein expression. In addition, cells treated with rapamycin showed a significant increase in p-Akt and a decrease in p-p70S6K that was associated with a decrease expression of vimentin and a slight increase expression in N-cadherin. On the other hand, cells treated with Akt inhibitor revealed a significant decrease in p-Akt and p-p70S6K that was associated with a significant decrease in vimentin and an increase in N-cadherin expression. In addition, cells transfected with DN-Akt or DN-S6K show significant increase expression in N-cadherin and a decrease in vimentin. Moreover, cells transfected with siRNA against rictor or siRNA against raptor resulted in a decrease in vimentin and an increase N-cadherin expression. Kidney tumors from TSC patients showed significant decrease in N-cadherin and significant increased in vimentin protein expression compared to control kidney tissues. These data comprise the first report to provide the role of Akt/tuberin/mTORC1/2 in the regulation of N-cadherin and vimentin that are involved in the progression of fibrosis in kidney tumor of TSC patients.

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Deficiency of tuberin results in increased vimentin protein expression in tumor kidney tissue of TSC patients(A) Representative Immunoblot analysis showed significant decreased in tuberin and increased in vimentin protein expression in tumor kidney (T) from patients with tuberous sclerosis compared to normal kidney tissues. Actin was used as loading control. (B) Histograms represent means±SE (n=6). Significant difference from control is indicated by **P<0.01. (C) H&E staining shows (a) a normal tubular and glomerular structure in control kidney tissue and (b) *fat, ^vessel and # smooth muscle cells types in kidney angiomyolipoma tissue of TSC patients. (D) Kidney sections were stained with vimentin followed by immunofluroscene staining. Control of kidney (c) shows a few cells stained with vimentin while (d) most of blood vessel and smooth muscle cells were stained in kidney tumor tissue. Control sections in both procedures were incubated without primary antibody.
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Figure 5: Deficiency of tuberin results in increased vimentin protein expression in tumor kidney tissue of TSC patients(A) Representative Immunoblot analysis showed significant decreased in tuberin and increased in vimentin protein expression in tumor kidney (T) from patients with tuberous sclerosis compared to normal kidney tissues. Actin was used as loading control. (B) Histograms represent means±SE (n=6). Significant difference from control is indicated by **P<0.01. (C) H&E staining shows (a) a normal tubular and glomerular structure in control kidney tissue and (b) *fat, ^vessel and # smooth muscle cells types in kidney angiomyolipoma tissue of TSC patients. (D) Kidney sections were stained with vimentin followed by immunofluroscene staining. Control of kidney (c) shows a few cells stained with vimentin while (d) most of blood vessel and smooth muscle cells were stained in kidney tumor tissue. Control sections in both procedures were incubated without primary antibody.

Mentions: To determine whether deficiency of tuberin in tumor tissue of TSC patients has an effect on vimentin protein expression, we extracted proteins from tumor kidney tissues of TSC patients and normal kidney tissues of healthy subjects. Western blot analysis were performed and blotted against tuberin and vimentin antibodies. Representative blots showed significant decreased expression of tuberin is associated with significant increase in vimentin expression in tumor kidney of TSC patients (Figure 5A&B). On the other hand, weak vimentin expression was detected in healthy subject of normal kidney tissues (Figure 5A&B). H&E staining showed normal tubular and glomerular structure in control kidney tissues, while fat, vessel and smooth muscle cells were shown in kidney section of angiomyolipoma tissue (Figure 5Ca&b). To confirm the expression of vimentin in kidney tissues from normal and tumor samples, immunofluorescence staining was performed. Data in Figure 5Dd showed a strong staining of vimentin in vessel and smooth muscle cells as well as around the fat cells in kidney section of angiomyolipoma tissue. While weak staining of vimentin was detected in normal tubular cells of kidney section of healthy subject (Figure 5Dc).


Tuberin-deficiency downregulates N-cadherin and upregulates vimentin in kidney tumor of TSC patients.

Liang S, Salas T, Gencaslan E, Li B, Habib SL - Oncotarget (2014)

Deficiency of tuberin results in increased vimentin protein expression in tumor kidney tissue of TSC patients(A) Representative Immunoblot analysis showed significant decreased in tuberin and increased in vimentin protein expression in tumor kidney (T) from patients with tuberous sclerosis compared to normal kidney tissues. Actin was used as loading control. (B) Histograms represent means±SE (n=6). Significant difference from control is indicated by **P<0.01. (C) H&E staining shows (a) a normal tubular and glomerular structure in control kidney tissue and (b) *fat, ^vessel and # smooth muscle cells types in kidney angiomyolipoma tissue of TSC patients. (D) Kidney sections were stained with vimentin followed by immunofluroscene staining. Control of kidney (c) shows a few cells stained with vimentin while (d) most of blood vessel and smooth muscle cells were stained in kidney tumor tissue. Control sections in both procedures were incubated without primary antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196174&req=5

Figure 5: Deficiency of tuberin results in increased vimentin protein expression in tumor kidney tissue of TSC patients(A) Representative Immunoblot analysis showed significant decreased in tuberin and increased in vimentin protein expression in tumor kidney (T) from patients with tuberous sclerosis compared to normal kidney tissues. Actin was used as loading control. (B) Histograms represent means±SE (n=6). Significant difference from control is indicated by **P<0.01. (C) H&E staining shows (a) a normal tubular and glomerular structure in control kidney tissue and (b) *fat, ^vessel and # smooth muscle cells types in kidney angiomyolipoma tissue of TSC patients. (D) Kidney sections were stained with vimentin followed by immunofluroscene staining. Control of kidney (c) shows a few cells stained with vimentin while (d) most of blood vessel and smooth muscle cells were stained in kidney tumor tissue. Control sections in both procedures were incubated without primary antibody.
Mentions: To determine whether deficiency of tuberin in tumor tissue of TSC patients has an effect on vimentin protein expression, we extracted proteins from tumor kidney tissues of TSC patients and normal kidney tissues of healthy subjects. Western blot analysis were performed and blotted against tuberin and vimentin antibodies. Representative blots showed significant decreased expression of tuberin is associated with significant increase in vimentin expression in tumor kidney of TSC patients (Figure 5A&B). On the other hand, weak vimentin expression was detected in healthy subject of normal kidney tissues (Figure 5A&B). H&E staining showed normal tubular and glomerular structure in control kidney tissues, while fat, vessel and smooth muscle cells were shown in kidney section of angiomyolipoma tissue (Figure 5Ca&b). To confirm the expression of vimentin in kidney tissues from normal and tumor samples, immunofluorescence staining was performed. Data in Figure 5Dd showed a strong staining of vimentin in vessel and smooth muscle cells as well as around the fat cells in kidney section of angiomyolipoma tissue. While weak staining of vimentin was detected in normal tubular cells of kidney section of healthy subject (Figure 5Dc).

Bottom Line: In addition, cells treated with rapamycin showed a significant increase in p-Akt and a decrease in p-p70S6K that was associated with a decrease expression of vimentin and a slight increase expression in N-cadherin.On the other hand, cells treated with Akt inhibitor revealed a significant decrease in p-Akt and p-p70S6K that was associated with a significant decrease in vimentin and an increase in N-cadherin expression.Kidney tumors from TSC patients showed significant decrease in N-cadherin and significant increased in vimentin protein expression compared to control kidney tissues.

View Article: PubMed Central - PubMed

Affiliation: South Texas Veterans Health Care System, San Antonio, TX.

ABSTRACT
Angiomyolipomas (AMLs) are associated with cell fibrosis in kidney of Tuberous Sclerosis Complex patients. The mechanism by which the fibrotic proteins accumulated in AMLs has not been explored. In the present study, we investigated the role of Akt/tuberin/mTOR pathway in the regulation cell fibrosis proteins. AML cells that expressed low levels of tuberin showed less expression of N-cadherin and higher of vimentin proteins compared to HEK293 cells. AML cells infected with Ad-tuberin showed a significant decrease in vimentin and an increase in N-cadherin protein expression. In addition, cells treated with rapamycin showed a significant increase in p-Akt and a decrease in p-p70S6K that was associated with a decrease expression of vimentin and a slight increase expression in N-cadherin. On the other hand, cells treated with Akt inhibitor revealed a significant decrease in p-Akt and p-p70S6K that was associated with a significant decrease in vimentin and an increase in N-cadherin expression. In addition, cells transfected with DN-Akt or DN-S6K show significant increase expression in N-cadherin and a decrease in vimentin. Moreover, cells transfected with siRNA against rictor or siRNA against raptor resulted in a decrease in vimentin and an increase N-cadherin expression. Kidney tumors from TSC patients showed significant decrease in N-cadherin and significant increased in vimentin protein expression compared to control kidney tissues. These data comprise the first report to provide the role of Akt/tuberin/mTORC1/2 in the regulation of N-cadherin and vimentin that are involved in the progression of fibrosis in kidney tumor of TSC patients.

Show MeSH
Related in: MedlinePlus