Journal
CELLS
Volume 10, Issue 2, Pages -Publisher
MDPI
DOI: 10.3390/cells10020243
Keywords
cutaneous squamous cell carcinoma; cancer stem cells; angiotensinogen; renin; prorenin receptor; angiotensin-converting enzyme; angiotensin-converting enzyme 2; angiotensin II receptor 1; angiotensin II receptor 2
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Funding
- Deane endowment Trust summer student scholarship
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Our study investigated the expression of renin-angiotensin system components by cancer stem cell subpopulations in metastatic head and neck cutaneous squamous cell carcinoma. Through immunohistochemical staining, Western blotting, and RT-qPCR, we confirmed the complex molecular expression pattern, with angiotensinogen, PRR, ACE, and AT(1)R transcripts present, while renin and AT(2)R were absent, and ACE2 expression was tissue-specific. Further immunostaining showed specific expression patterns within tumor nests and peritumoral stroma.
We investigated the expression of components of the renin-angiotensin system (RAS) by cancer stem cell (CSC) subpopulations in metastatic head and neck cutaneous squamous cell carcinoma (mHNcSCC). Immunohistochemical staining demonstrated expression of prorenin receptor (PRR), angiotensin-converting enzyme (ACE), and angiotensin II receptor 2 (AT(2)R) in all cases and angiotensinogen in 14 cases; however, renin and ACE2 were not detected in any of the 20 mHNcSCC tissue samples. Western blotting showed protein expression of angiotensinogen in all six mHNcSCC tissue samples, but in none of the four mHNcSCC-derived primary cell lines, while PRR was detected in the four cell lines only. RT-qPCR confirmed transcripts of angiotensinogen, PRR, ACE, and angiotensin II receptor 1 (AT(1)R), but not renin or AT(2)R in all four mHNcSCC tissue samples and all four mHNcSCC-derived primary cell lines, while ACE2 was expressed in the tissue samples only. Double immunohistochemical staining on two of the mHNcSCC tissue samples showed expression of angiotensinogen by the SOX2+ CSCs within the tumor nests (TNs), and immunofluorescence showed expression of PRR and AT(2)R by the SOX2+ CSCs within the TNs and the peritumoral stroma (PTS). ACE was expressed on the endothelium of the tumor microvessels within the PTS. We demonstrated expression of angiotensinogen by CSCs within the TNs, PRR, and AT(2)R by the CSCs within the TNs and the PTS, in addition to ACE on the endothelium of tumor microvessels in mHNcSCC.
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