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Cancer Stem Cells in Moderately Differentiated Buccal Mucosal Squamous Cell Carcinoma Express Components of the Renin – Angiotensin System

View Article: PubMed Central - PubMed

ABSTRACT

Aim: We have recently identified and characterized cancer stem cell (CSC) subpopulations within moderately differentiated buccal mucosal squamous cell carcinoma (MDBMSCC). We hypothesized that these CSCs express components of the renin–angiotensin system (RAS).

Methods: 3,3′-Diaminobenzidine (DAB) immunohistochemical (IHC) staining was performed on formalin-fixed paraffin-embedded MDBMSCC samples to investigate the expression of the components of the RAS: (pro)renin receptor (PRR), angiotensin converting enzyme (ACE), angiotensin II receptor 1 (ATIIR1), and angiotensin II receptor 2 (ATIIR2). NanoString mRNA gene expression analysis and Western Blotting (WB) were performed on snap-frozen MDBMSCC samples to confirm gene expression and translation of these transcripts, respectively. Double immunofluorescent (IF) IHC staining of these components of the RAS with the embryonic stem cell markers OCT4 or SALL4 was performed to demonstrate their localization in relation to the CSC subpopulations within MDBMSCC.

Results: DAB IHC staining demonstrated expression of PRR, ACE, ATIIR1, and ATIIR2 in MDBMSCC. IF IHC staining showed that PRR was expressed by the CSC subpopulations within the tumor nests, the peri-tumoral stroma, and the endothelium of the microvessels within the peri-tumoral stroma. ATIIR1 and ATIIR2 were localized to the CSC subpopulations within the tumor nests and the peri-tumoral stroma, while ACE was localized to the endothelium of the microvessels within the peri-tumoral stroma. WB and NanoString analyses confirmed protein expression and transcription activation of PRR, ACE, and ATIIR1, but not of ATIIR2, respectively.

Conclusion: Our novel findings of the presence and localization of PRR, ACE, ATIIR1, and potentially ATIIR2 to the CSC subpopulations within MDBMSCC suggest CSC as a therapeutic target by modulation of the RAS.

No MeSH data available.


Related in: MedlinePlus

Relative expression of mRNA transcripts of the components of the renin–angiotensin system in five moderately differentiated buccal mucosal squamous cell carcinoma samples. Expression is depicted in relative units (RU) as a ratio to the GUSB housekeeper. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples.
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Figure 4: Relative expression of mRNA transcripts of the components of the renin–angiotensin system in five moderately differentiated buccal mucosal squamous cell carcinoma samples. Expression is depicted in relative units (RU) as a ratio to the GUSB housekeeper. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples.

Mentions: NanoString gene analysis of the components of the RAS: PRR, ACE, ATIIR1, and ATIIR2, was performed in five samples of MDBMSCC from the original cohort of six patients used for DAB IHC staining, normalized against the housekeeping gene, GUSB. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples (Figure 4).


Cancer Stem Cells in Moderately Differentiated Buccal Mucosal Squamous Cell Carcinoma Express Components of the Renin – Angiotensin System
Relative expression of mRNA transcripts of the components of the renin–angiotensin system in five moderately differentiated buccal mucosal squamous cell carcinoma samples. Expression is depicted in relative units (RU) as a ratio to the GUSB housekeeper. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037224&req=5

Figure 4: Relative expression of mRNA transcripts of the components of the renin–angiotensin system in five moderately differentiated buccal mucosal squamous cell carcinoma samples. Expression is depicted in relative units (RU) as a ratio to the GUSB housekeeper. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples.
Mentions: NanoString gene analysis of the components of the RAS: PRR, ACE, ATIIR1, and ATIIR2, was performed in five samples of MDBMSCC from the original cohort of six patients used for DAB IHC staining, normalized against the housekeeping gene, GUSB. Transcriptional profiling confirmed the presence of PRR and ACE mRNA in all five samples and ATIIR1 in one sample, but ATIIR2 mRNA was below the detectable level within all five samples (Figure 4).

View Article: PubMed Central - PubMed

ABSTRACT

Aim: We have recently identified and characterized cancer stem cell (CSC) subpopulations within moderately differentiated buccal mucosal squamous cell carcinoma (MDBMSCC). We hypothesized that these CSCs express components of the renin–angiotensin system (RAS).

Methods: 3,3′-Diaminobenzidine (DAB) immunohistochemical (IHC) staining was performed on formalin-fixed paraffin-embedded MDBMSCC samples to investigate the expression of the components of the RAS: (pro)renin receptor (PRR), angiotensin converting enzyme (ACE), angiotensin II receptor 1 (ATIIR1), and angiotensin II receptor 2 (ATIIR2). NanoString mRNA gene expression analysis and Western Blotting (WB) were performed on snap-frozen MDBMSCC samples to confirm gene expression and translation of these transcripts, respectively. Double immunofluorescent (IF) IHC staining of these components of the RAS with the embryonic stem cell markers OCT4 or SALL4 was performed to demonstrate their localization in relation to the CSC subpopulations within MDBMSCC.

Results: DAB IHC staining demonstrated expression of PRR, ACE, ATIIR1, and ATIIR2 in MDBMSCC. IF IHC staining showed that PRR was expressed by the CSC subpopulations within the tumor nests, the peri-tumoral stroma, and the endothelium of the microvessels within the peri-tumoral stroma. ATIIR1 and ATIIR2 were localized to the CSC subpopulations within the tumor nests and the peri-tumoral stroma, while ACE was localized to the endothelium of the microvessels within the peri-tumoral stroma. WB and NanoString analyses confirmed protein expression and transcription activation of PRR, ACE, and ATIIR1, but not of ATIIR2, respectively.

Conclusion: Our novel findings of the presence and localization of PRR, ACE, ATIIR1, and potentially ATIIR2 to the CSC subpopulations within MDBMSCC suggest CSC as a therapeutic target by modulation of the RAS.

No MeSH data available.


Related in: MedlinePlus