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Proteomic analysis of bladder cancer indicates Prx-I as a key molecule in BI-TK/GCV treatment system.

Jiang L, Xiao X, Ren J, Tang Y, Weng H, Yang Q, Wu M, Tang W - PLoS ONE (2014)

Bottom Line: In order to understand the molecular mechanisms of Bifidobacterium infantis thymidine kinase/nucleoside analogue ganciclovir (BI-TK/GCV) treatment system which was proven to exhibit sustainable anti-tumor growth activity and induce apoptosis in bladder cancer, a proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ), followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. 192 down-regulated and 210 up-regulated proteins were identified after treatment with BI-TK/GCV system in Sprague-Dawley (SD) rats.Western blot analysis and immunohistochemistry analysis confirmed that Peroxiredoxin-I (Prx-I) was significantly down-regulated in bladder cancer after treatment.Prx-I silencing by transfection of Prx-I shRNA significantly suppressed growth, promoted apoptosis and regulated the cell cycle in T24 cells and reduced the phospho-NF-κB p50 and p65 protein expression which revealed the links between Prx-I and NF-κB pathway implied by Ingenuity pathway analysis (IPA).

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

ABSTRACT
In order to understand the molecular mechanisms of Bifidobacterium infantis thymidine kinase/nucleoside analogue ganciclovir (BI-TK/GCV) treatment system which was proven to exhibit sustainable anti-tumor growth activity and induce apoptosis in bladder cancer, a proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ), followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. 192 down-regulated and 210 up-regulated proteins were identified after treatment with BI-TK/GCV system in Sprague-Dawley (SD) rats. Western blot analysis and immunohistochemistry analysis confirmed that Peroxiredoxin-I (Prx-I) was significantly down-regulated in bladder cancer after treatment. Prx-I silencing by transfection of Prx-I shRNA significantly suppressed growth, promoted apoptosis and regulated the cell cycle in T24 cells and reduced the phospho-NF-κB p50 and p65 protein expression which revealed the links between Prx-I and NF-κB pathway implied by Ingenuity pathway analysis (IPA). These findings yield new insights into the therapy of bladder cancer, revealing Prx-I as a new therapeutic target and indicating BI-TK/GCV system as a prospective therapy by down-regulation of Prx-I through NF-κB signaling pathway.

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Related in: MedlinePlus

Validation of Prx-I by Western blot and immunohistochemical (IHC) analysis.A: Expressions of Prx-I in four groups by Western blot analysis. Beta-actin was used as a loading control. B: Representative images showing the immunoexpression of Prx-I in tumor tissues of four groups. Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. (Asterisk (*) indicates P<0.05 in BI-TK group versus normal saline group)
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pone-0098764-g003: Validation of Prx-I by Western blot and immunohistochemical (IHC) analysis.A: Expressions of Prx-I in four groups by Western blot analysis. Beta-actin was used as a loading control. B: Representative images showing the immunoexpression of Prx-I in tumor tissues of four groups. Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. (Asterisk (*) indicates P<0.05 in BI-TK group versus normal saline group)

Mentions: The differential expression levels of Prx- I identified by iTRAQ approach were validated by Western blot (Figure 3A). Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. Moreover, the Prx-I expression in the tumor tissues was verified by IHC analysis (Figure 3B). The Prx-I content in bladder cancer cells of BI-TK group was significantly lower than that in the other groups (p<0.05, Figure 3B), which is consistent with the results obtained by iTRAQ and Western blot.


Proteomic analysis of bladder cancer indicates Prx-I as a key molecule in BI-TK/GCV treatment system.

Jiang L, Xiao X, Ren J, Tang Y, Weng H, Yang Q, Wu M, Tang W - PLoS ONE (2014)

Validation of Prx-I by Western blot and immunohistochemical (IHC) analysis.A: Expressions of Prx-I in four groups by Western blot analysis. Beta-actin was used as a loading control. B: Representative images showing the immunoexpression of Prx-I in tumor tissues of four groups. Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. (Asterisk (*) indicates P<0.05 in BI-TK group versus normal saline group)
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048271&req=5

pone-0098764-g003: Validation of Prx-I by Western blot and immunohistochemical (IHC) analysis.A: Expressions of Prx-I in four groups by Western blot analysis. Beta-actin was used as a loading control. B: Representative images showing the immunoexpression of Prx-I in tumor tissues of four groups. Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. (Asterisk (*) indicates P<0.05 in BI-TK group versus normal saline group)
Mentions: The differential expression levels of Prx- I identified by iTRAQ approach were validated by Western blot (Figure 3A). Compared with the normal saline group, expression of Prx-I is down-regulated in the other three groups (especially in the BI-TK group), which is similar to the results obtained by iTRAQ. Moreover, the Prx-I expression in the tumor tissues was verified by IHC analysis (Figure 3B). The Prx-I content in bladder cancer cells of BI-TK group was significantly lower than that in the other groups (p<0.05, Figure 3B), which is consistent with the results obtained by iTRAQ and Western blot.

Bottom Line: In order to understand the molecular mechanisms of Bifidobacterium infantis thymidine kinase/nucleoside analogue ganciclovir (BI-TK/GCV) treatment system which was proven to exhibit sustainable anti-tumor growth activity and induce apoptosis in bladder cancer, a proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ), followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. 192 down-regulated and 210 up-regulated proteins were identified after treatment with BI-TK/GCV system in Sprague-Dawley (SD) rats.Western blot analysis and immunohistochemistry analysis confirmed that Peroxiredoxin-I (Prx-I) was significantly down-regulated in bladder cancer after treatment.Prx-I silencing by transfection of Prx-I shRNA significantly suppressed growth, promoted apoptosis and regulated the cell cycle in T24 cells and reduced the phospho-NF-κB p50 and p65 protein expression which revealed the links between Prx-I and NF-κB pathway implied by Ingenuity pathway analysis (IPA).

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

ABSTRACT
In order to understand the molecular mechanisms of Bifidobacterium infantis thymidine kinase/nucleoside analogue ganciclovir (BI-TK/GCV) treatment system which was proven to exhibit sustainable anti-tumor growth activity and induce apoptosis in bladder cancer, a proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ), followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. 192 down-regulated and 210 up-regulated proteins were identified after treatment with BI-TK/GCV system in Sprague-Dawley (SD) rats. Western blot analysis and immunohistochemistry analysis confirmed that Peroxiredoxin-I (Prx-I) was significantly down-regulated in bladder cancer after treatment. Prx-I silencing by transfection of Prx-I shRNA significantly suppressed growth, promoted apoptosis and regulated the cell cycle in T24 cells and reduced the phospho-NF-κB p50 and p65 protein expression which revealed the links between Prx-I and NF-κB pathway implied by Ingenuity pathway analysis (IPA). These findings yield new insights into the therapy of bladder cancer, revealing Prx-I as a new therapeutic target and indicating BI-TK/GCV system as a prospective therapy by down-regulation of Prx-I through NF-κB signaling pathway.

Show MeSH
Related in: MedlinePlus