Identification of Cancer Stem Cell Subpopulations in Head and Neck Metastatic Malignant Melanoma.

Vithushiya Yoganandarajah,Swee T Tan,Josie Patel,Imogen M Roth,Helen D Brasch,Dalice Sim,Erin Paterson,Nicholas Bockett,Bede Van Schaijik,Paul F Davis,Tinte Itinteang

doi:10.3390/cells9020324

Vithushiya Yoganandarajah, Swee T Tan + Show 9 more

Open Access

https://doi.org/10.3390/cells9020324

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Abstract

Cancer stem cells (CSCs) have been identified in many cancer types. This study identified and characterized CSCs in head and neck metastatic malignant melanoma (HNmMM) to regional lymph nodes using induced pluripotent stem cell (iPSC) markers. Immunohistochemical (IHC) staining performed on 20 HNmMM tissue samples demonstrated expression of iPSC markers OCT4, SOX2, KLF4, and c-MYC in all samples, while NANOG was expressed at low levels in two samples. Immunofluorescence (IF) staining demonstrated an OCT4+/SOX2+/KLF4+/c-MYC+ CSC subpopulation within the tumor nests (TNs) and another within the peritumoral stroma (PTS) of HNmMM tissues. IF also showed expression of NANOG by some OCT4+/SOX2+/KLF4+/c-MYC+ cells within the TNs in an HNmMM tissue sample that expressed NANOG on IHC staining. In situ hybridization (n = 6) and reverse-transcription quantitative polymerase chain reaction (n = 5) on the HNmMM samples confirmed expression of all five iPSC markers. Western blotting of primary cell lines derived from four of the 20 HNmMM tissue samples showed expression of SOX2, KLF4, and c-MYC but not OCT4 and NANOG, and three of these cell lines formed tumorspheres in vitro. We demonstrate the presence of two putative CSC subpopulations within HNmMM, which may be a novel therapeutic target in the treatment of this aggressive cancer.

Highlights

In 2018, 287,723 new cases of malignant melanoma (MM) were diagnosed worldwide with a record of 60,712 deaths [1]
octamer-binding transcription factor 4 (OCT4), NANOG, sex-determining region Y-box 2 (SOX2), Kruppel-like factor 4 (KLF4), and c-MYC Proteins are Expressed in head and neck metastatic malignant melanoma (HNmMM) Tissue Samples
In order to confirm the presence of HNmMM, the slides were stained with H&E that showed tumor nests (TNs), surrounded by peritumoral stroma (PTS) in all 20 tissue samples (Figure ??A)

Summary

Introduction

In 2018, 287,723 new cases of malignant melanoma (MM) were diagnosed worldwide with a record of 60,712 deaths [1]. MM accounts for 60–80% of deaths from all skin cancers globally with. New Zealand and Australia having the highest incidence [2,3,4]. The 5-year survival of metastatic MM (mMM) is 5–19% depending on the site of the metastasis [2,5] with an overall median survival of. Sentinel lymph node biopsy confers prognostic value and a potential survival advantage for patients with nodal metastases from intermediate-thickness MM who are treated with elective nodal dissection [8,9]. Surgical excision and adjuvant radiotherapy are the mainstay treatment for nodal mMM, with distant metastases requiring chemotherapy and/or immunotherapy [8,9].

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