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Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin ‚Äď Angiotensin System

View Article: PubMed Central - PubMed

ABSTRACT

Aim: To investigate the expression of the renin–angiotensin system (RAS) in cancer stem cells (CSCs), we have previously characterized in glioblastoma multiforme (GBM).

Methods: 3,3-Diaminobenzidine (DAB) immunohistochemical (IHC) staining for the stem cell marker, SOX2, and components of the RAS: angiotensin converting enzyme (ACE), (pro)renin receptor (PRR), angiotensin II receptor 1 (ATIIR1), and angiotensin II receptor 2 (ATIIR2) on 4 μm-thick formalin-fixed paraffin-embedded sections of previously characterized GBM samples in six patients was undertaken. Immunofluorescent (IF) IHC staining was performed to demonstrate expression of GFAP, SOX2, PRR, ACE, ATIIR1, and ATIIR2. The protein expression and the transcriptional activities of the genes encoding for ACE, PRR, ATIIR1, and ATIIR2 were studied using Western blotting (WB) and NanoString gene expression analysis, respectively.

Results: DAB and IF IHC staining demonstrated the expression SOX2 on the GFAP+ GBM CSCs. Cytoplasmic expression of PRR by the GFAP+ CSCs and the endothelium of the microvessels was observed. ACE was expressed on the endothelium of the microvessels only, while nuclear and cytoplasmic expression of ATIIR1 and ATIIR2 was observed on the endothelium of the microvessels and the CSCs. ATIIR1 was expressed on the GFAP+ CSCs cells, and ATIIR2 was expressed by the SOX2+ CSCs. The expression of ACE, PRR, and ATIIR1, but not ATIIR2, was confirmed by WB. NanoString gene analysis demonstrated transcriptional activation of ACE, PRR, and ATIIR1, but not ATIIR2.

Conclusion: This study demonstrated the expression of PRR, ATIIR1, and ATIIR2 by the SOX2 CSC population, and ACE on the endothelium of the microvessels, within GBM. ACE, PRR, and ATIIR1 were expressed at the protein and mRNA levels, with ATIIR2 detectable only by IHC staining. This novel finding suggests that the CSCs may be a novel therapeutic target for GBM by modulation of the RAS.

No MeSH data available.


Western blots demonstrating the expression of PRR (~38‚ÄČkDa) (A) and ATIIR1 (~45‚ÄČkDa) (B) in all five GBM samples. ATIIR2 was not detected in any of the samples (C). ACE was detected in four out of the five GBM samples examined (D).
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Figure 3: Western blots demonstrating the expression of PRR (~38‚ÄČkDa) (A) and ATIIR1 (~45‚ÄČkDa) (B) in all five GBM samples. ATIIR2 was not detected in any of the samples (C). ACE was detected in four out of the five GBM samples examined (D).

Mentions: Western blotting was performed to examine the presence of components of the RAS in GBM samples of five patients included in DAB IHC staining. PRR (Figure 3A) and ATIIR1 (Figure 3B) were present in all five samples with bands of ~37 and 45‚ÄČkDa, respectively. Bands of ~70‚ÄČkDa represent PRR dimerization (Figure 3A). ATIIR2 was absent in all five samples (Figure 3C), while ACE was present, at low levels, in all five samples (Figure 3D).


Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin ‚Äď Angiotensin System
Western blots demonstrating the expression of PRR (~38‚ÄČkDa) (A) and ATIIR1 (~45‚ÄČkDa) (B) in all five GBM samples. ATIIR2 was not detected in any of the samples (C). ACE was detected in four out of the five GBM samples examined (D).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037176&req=5

Figure 3: Western blots demonstrating the expression of PRR (~38‚ÄČkDa) (A) and ATIIR1 (~45‚ÄČkDa) (B) in all five GBM samples. ATIIR2 was not detected in any of the samples (C). ACE was detected in four out of the five GBM samples examined (D).
Mentions: Western blotting was performed to examine the presence of components of the RAS in GBM samples of five patients included in DAB IHC staining. PRR (Figure 3A) and ATIIR1 (Figure 3B) were present in all five samples with bands of ~37 and 45‚ÄČkDa, respectively. Bands of ~70‚ÄČkDa represent PRR dimerization (Figure 3A). ATIIR2 was absent in all five samples (Figure 3C), while ACE was present, at low levels, in all five samples (Figure 3D).

View Article: PubMed Central - PubMed

ABSTRACT

Aim: To investigate the expression of the renin–angiotensin system (RAS) in cancer stem cells (CSCs), we have previously characterized in glioblastoma multiforme (GBM).

Methods: 3,3-Diaminobenzidine (DAB) immunohistochemical (IHC) staining for the stem cell marker, SOX2, and components of the RAS: angiotensin converting enzyme (ACE), (pro)renin receptor (PRR), angiotensin II receptor 1 (ATIIR1), and angiotensin II receptor 2 (ATIIR2) on 4 μm-thick formalin-fixed paraffin-embedded sections of previously characterized GBM samples in six patients was undertaken. Immunofluorescent (IF) IHC staining was performed to demonstrate expression of GFAP, SOX2, PRR, ACE, ATIIR1, and ATIIR2. The protein expression and the transcriptional activities of the genes encoding for ACE, PRR, ATIIR1, and ATIIR2 were studied using Western blotting (WB) and NanoString gene expression analysis, respectively.

Results: DAB and IF IHC staining demonstrated the expression SOX2 on the GFAP+ GBM CSCs. Cytoplasmic expression of PRR by the GFAP+ CSCs and the endothelium of the microvessels was observed. ACE was expressed on the endothelium of the microvessels only, while nuclear and cytoplasmic expression of ATIIR1 and ATIIR2 was observed on the endothelium of the microvessels and the CSCs. ATIIR1 was expressed on the GFAP+ CSCs cells, and ATIIR2 was expressed by the SOX2+ CSCs. The expression of ACE, PRR, and ATIIR1, but not ATIIR2, was confirmed by WB. NanoString gene analysis demonstrated transcriptional activation of ACE, PRR, and ATIIR1, but not ATIIR2.

Conclusion: This study demonstrated the expression of PRR, ATIIR1, and ATIIR2 by the SOX2 CSC population, and ACE on the endothelium of the microvessels, within GBM. ACE, PRR, and ATIIR1 were expressed at the protein and mRNA levels, with ATIIR2 detectable only by IHC staining. This novel finding suggests that the CSCs may be a novel therapeutic target for GBM by modulation of the RAS.

No MeSH data available.