Gillies McIndoe Research Institute, PO Box 7184, Newtown, Wellington 6242, New Zealand.
Gillies McIndoe Research Institute, PO Box 7184, Newtown, Wellington 6242, New Zealand; Auckland University, Private Bag 92019, Victoria St, West Auckland 1142, New Zealand.
J Plast Reconstr Aesthet Surg. 2019 Sep;72(9):1484-1493. doi: 10.1016/j.bjps.2018.11.013. Epub 2018 Nov 22.
To investigate the expression of components of the renin-angiotensin system (RAS): pro-renin receptor (PRR), angiotensin converting enzyme (ACE), angiotensin II receptor 1 (ATIIR1) and angiotensin II receptor 2 (ATIIR2) by the cancer stem cell (CSC) subpopulations in moderately differentiated head and neck cutaneous squamous cell carcinoma (MDHNCSCC).
3,3-Diaminobenzidine (DAB) immunohistochemical (IHC) staining for PRR, ACE, ATIIR1 and ATIIR2 was performed on formalin-fixed paraffin-embedded sections of ten MDHNCSCC tissue samples. Immunofluorescence (IF) IHC staining was used to localise components of the RAS. Western blotting (WB) and RT-qPCR were performed on snap-frozen MDHNCSCC tissue samples and MDHNCSCC-derived primary cell lines to investigate protein transcription expression of these proteins, respectively.
DAB IHC staining demonstrated the presence of PRR, ACE, ATIIR1 and ATIIR2 in all ten MDHNCSCC tissue samples. IF IHC staining showed expression of PRR and ATIIR2 by the OCT4 cells, and ACE and ATIIR1 by the SOX2 cells, within the tumour nests (TNs) and the peritumoural stroma (PTS). PRR, ACE, ATIIR1 and ATIIR2 were expressed by the endothelium of the microvessels within the PTS. WB confirmed protein expression for PRR, ACE and ATIIR1 in MDHNCSCC tissue samples and MDHNCSCC-derived primary cell lines. RT-qPCR showed transcriptional activation of PRR, ACE, ATIIR1 and ATIIR2 in MDHNCSCC tissue samples; and PRR, ACE, ATIIR1 but not ATIIR2, in MDHNCSCC-derived primary cell lines.
PRR, ACE, ATIIR1 and ATIIR2 are expressed by the CSC subpopulations within the TNs, the PTS, and the endothelium of the microvessels within the PTS, in MDHNCSCC.
研究头颈部皮肤鳞癌（MDHNCSCC）中等分化样本中，癌症干细胞（CSC）亚群中肾素-血管紧张素系统（RAS）的组成部分：前肾素受体（PRR）、血管紧张素转换酶（ACE）、血管紧张素 II 受体 1（ATIIR1）和血管紧张素 II 受体 2（ATIIR2）的表达。
对 10 例 MDHNCSCC 组织样本的福尔马林固定石蜡包埋切片进行 3,3-二氨基联苯胺（DAB）免疫组织化学（IHC）染色，以检测 PRR、ACE、ATIIR1 和 ATIIR2。免疫荧光（IF）IHC 染色用于定位 RAS 的组成部分。对 MDHNCSCC 组织样本和 MDHNCSCC 来源的原代细胞系进行 snap-frozen Western 印迹（WB）和 RT-qPCR，以分别研究这些蛋白的蛋白转录表达。
DAB IHC 染色显示 10 例 MDHNCSCC 组织样本中均存在 PRR、ACE、ATIIR1 和 ATIIR2。IF IHC 染色显示，在肿瘤巢（TN）和肿瘤周围基质（PTS）中，OCT4 细胞表达 PRR 和 ATIIR2，SOX2 细胞表达 ACE 和 ATIIR1。PRR、ACE、ATIIR1 和 ATIIR2 也表达于 PTS 中小血管的内皮细胞中。WB 证实 MDHNCSCC 组织样本和 MDHNCSCC 来源的原代细胞系中存在 PRR、ACE 和 ATIIR1 蛋白表达。RT-qPCR 显示 PRR、ACE、ATIIR1 和 ATIIR2 在 MDHNCSCC 组织样本中均有转录激活；而在 MDHNCSCC 来源的原代细胞系中，只有 PRR、ACE、ATIIR1 有转录激活，而 ATIIR2 没有。
在 MDHNCSCC 中，PRR、ACE、ATIIR1 和 ATIIR2 由 TN 内、PTS 内和 PTS 中小血管内皮细胞中的 CSC 亚群表达。
