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

2019 ◽  
Author(s):  
Vithushiya Yoganandarajah ◽  
Josie Patel ◽  
Bede van Schaijik ◽  
Nicholas Bockett ◽  
Helen D. Brasch ◽  
...  
Keyword(s):  
Stem Cell ◽  
Malignant Melanoma ◽  
Head And Neck ◽  
Cell Lines ◽  
Tissue Sample ◽  
Quantitative Polymerase Chain Reaction ◽  
Metastatic Malignant Melanoma ◽  
Regional Lymph Nodes ◽  
Tissue Samples ◽  
Two Samples

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 four 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.

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

Cells ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 324 ◽  
Author(s):  
Vithushiya Yoganandarajah ◽  
Josie Patel ◽  
Bede van Schaijik ◽  
Nicholas Bockett ◽  
Helen D. Brasch ◽  
...  
Keyword(s):  
Stem Cell ◽  
Malignant Melanoma ◽  
Head And Neck ◽  
Cell Lines ◽  
Tissue Sample ◽  
Quantitative Polymerase Chain Reaction ◽  
Metastatic Malignant Melanoma ◽  
Regional Lymph Nodes ◽  
Tissue Samples ◽  
Two Samples

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.

scholarly journals Cancer Stem Cells in Head and Neck Metastatic Malignant Melanoma Express Components of the Renin-Angiotensin System

Life ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 268
Author(s):  
Sam Siljee ◽  
Tessa Pilkington ◽  
Helen D. Brasch ◽  
Nicholas Bockett ◽  
Josie Patel ◽  
...  
Keyword(s):  
Stem Cells ◽  
Malignant Melanoma ◽  
Cancer Stem Cells ◽  
Cell Lines ◽  
Renin Angiotensin System ◽  
Metastatic Malignant Melanoma ◽  
Primary Cell ◽  
Tissue Samples ◽  
Angiotensin System ◽  
Primary Cell Lines

Components of the renin-angiotensin system (RAS) are expressed by cancer stem cells (CSCs) in many cancer types. We here investigated expression of the RAS by the CSC subpopulations in human head and neck metastatic malignant melanoma (HNmMM) tissue samples and HNmMM-derived primary cell lines. Immunohistochemical staining demonstrated expression of pro-renin receptor (PRR), angiotensin-converting enzyme (ACE), and angiotensin II receptor 2 (AT2R) in all; renin in one; and ACE2 in none of the 20 HNmMM tissue samples. PRR was localized to cells within the tumor nests (TNs), while AT2R was expressed by cells within the TNs and the peritumoral stroma (PTS). ACE was localized to the endothelium of the tumor microvessels within the PTS. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) detected transcripts for PRR, ACE, ACE2, and AT1R, in all the five HNmMM tissue samples and four HNmMM-derived primary cell lines; renin in one tissue sample and one cell line, and AT2R in none of the five HNmMM tissue samples and cell lines. Western blotting showed variable expression of ACE, PRR, and AT2R, but not ACE2, in six HNmMM tissue samples and two HNmMM-derived primary cell lines. Immunofluorescence staining of two HNmMM tissue samples demonstrated expression of PRR and AT2R by the SOX2+ CSCs within the TNs and the OCT4+ CSCs within the PTS, with ACE localized to the endothelium of the tumor microvessels within the PTS.

scholarly journals Cancer stem cells enrichment with surface markers CD271 and CD44 in human head and neck squamous cell carcinomas

2019 ◽  
Vol 41 (4) ◽  
pp. 458-466 ◽  
Author(s):  
Osama A Elkashty ◽  
Ghada Abu Elghanam ◽  
Xinyun Su ◽  
Younan Liu ◽  
Peter J Chauvin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Head And Neck ◽  
Cell Lines ◽  
Squamous Cell ◽  
Colony Formation ◽  
Self Renewal ◽  
Tissue Samples ◽  
Hnscc Cell

Abstract Head and neck squamous cell carcinoma (HNSCC) has a poor 5-year survival rate of 50%. One potential reason for treatment failure is the presence of cancer stem cells (CSCs). Several cell markers, particularly CD44, have been used to isolate CSCs. However, isolating a pure population of CSC in HNSCC still remains a challenging task. Recent findings show that normal oral stem cells were isolated using CD271 as a marker. Thus, we investigated the combined use of CD271 and CD44 to isolate an enriched subpopulation of CSCs, followed by their characterization in vitro, in vivo, and in patients’ tissue samples. Fluorescent-activated cell sorting was used to isolate CD44+/CD271+ and CD44+/CD271− from two human HNSCC cell lines. Cell growth and self-renewal were measured with MTT and sphere/colony formation assays. Treatment-resistance was tested against chemotherapy (cisplatin and 5-fluorouracil) and ionizing radiation. Self-renewal, resistance, and stemness-related genes expression were measured with qRT-PCR. In vivo tumorigenicity was tested with an orthotopic immunodeficient mouse model of oral cancer. Finally, we examined the co-localization of CD44+/CD271+ in patients’ tissue samples. We found that CD271+ cells were a subpopulation of CD44+ cells in human HNSCC cell lines and tissues. CD44+/CD271+ cells exhibited higher cell proliferation, sphere/colony formation, chemo- and radio-resistance, upregulation of CSCs-related genes, and in vivo tumorigenicity when compared to CD44+/CD271− or the parental cell line. These cell markers showed increased expression in patients with the increase of the tumor stage. In conclusion, using both CD44 and CD271 allowed the isolation of CSCs from HNSCC. These enriched CSCs will be more relevant in future treatment and HNSCC progression studies.

scholarly journals Metastatic Malignant Melanoma of Parotid Gland with a Regressed Primary Tumor

2016 ◽  
Vol 2016 ◽  
pp. 1-4
Author(s):  
M. Mustafa Kılıçkaya ◽  
Giray Aynali ◽  
Ali Murat Ceyhan ◽  
Metin Çiriş
Keyword(s):  
Malignant Melanoma ◽  
Parotid Gland ◽  
Head And Neck ◽  
Primary Tumor ◽  
Primary Melanoma ◽  
Metastatic Malignant Melanoma ◽  
Malignant Melanomas ◽  
Pigmented Lesions

Malignant melanoma of the parotid gland is often metastatic and mainly originates from malignant melanomas in the head and neck. Nevertheless, some malignant melanomas may metastasize and subsequently regress. Therefore, it may not be possible to observe a metastatic malignant melanoma and its primary melanoma simultaneously. The investigation of a patient’s old photographs may help in the detection of preexisting and regressed pigmented lesions in the facial and neck regions.

scholarly journals Investigation of the expression of RIF1 gene on head and neck, pancreatic and brain cancer and cancer stem cells

2016 ◽  
Vol 39 (6) ◽  
pp. 43 ◽  
Author(s):  
Hacer E GursesCila ◽  
Muradiye Acar ◽  
Furkan B Barut ◽  
Mehmet Gunduz ◽  
Reidar Grenman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pancreatic Cancer ◽  
Stem Cells ◽  
Stem Cell ◽  
Cancer Stem Cells ◽  
Head And Neck ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines

Purpose: Recent studies have shown that cancer stem cells are resistant to chemotherapy. The aim of this study was to compare RIF1 gene expression in head and neck, pancreatic cancer and glioma cell lines and the cancer stem cells isolated from these cell lines. Methods: UT-SCC-74 from Turku University and UT-SCC-74B primary tumor metastasis and neck cancer cell lines, YKG-1 glioma cancer cell line from RIKEN, pancreatic cancer cell lines and ASPC-1 cells from ATCC were grown in cell culture. To isolate cancer stem cells, ALDH-1 for UT-SCC-74 and UT-SCC-74B cell line, CD-133 for YKG-1 cell line and CD-24 for ASPC-1 cell line, were used as markers of cancer stem cells. RNA isolation was performed for both cancer lines and cancer stem cells. RNAs were converted to cDNA. RIF1 gene expression was performed by qRT-PCR analysis. RIF1 gene expression was compared with cancer cell lines and cancer stem cells isolated from these cell lines. The possible effect of RIF1 gene was evaluated. Results: In the pancreatic cells, RIF1 gene expression in the stem cell-positive cell line was 256 time that seen in the stem cell-negative cell line. Conclusion: Considering the importance of RIF1 in NHEJ and of NHEJ in pancreatic cancer, RIF1 may be one of the genes that plays an important role in the diagnoses and therapeutic treatment of pancreatic cancer. The results of head and neck and brain cancers are inconclusive and further studies are required to elucidate the connection between RIF1 gene and these other types of cancers.

scholarly journals Effects of active acromegaly on bone mRNA and microRNA expression patterns

2018 ◽  
Vol 178 (4) ◽  
pp. 353-364 ◽  
Author(s):  
Zhanna Belaya ◽  
Tatiana Grebennikova ◽  
Galina Melnichenko ◽  
Alexey Nikitin ◽  
Alexander Solodovnikov ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Bone Tissue ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Sella Turcica ◽  
Compensatory Mechanisms ◽  
Tissue Samples ◽  
Active Acromegaly ◽  
Cell Commitment

Objective To evaluate the response of bone to chronic long-term growth hormone (GH) and insulin-like growth factor-1 (IGF1) excess by measuring the expression of selected mRNA and microRNA (miR) in bone tissue samples of patients with active acromegaly. Design Case–control study. Methods Bone tissue samples were obtained during transsphenoidal adenomectomy from the sphenoid bone (sella turcica) from 14 patients with clinically and biochemically confirmed acromegaly and 10 patients with clinically non-functioning pituitary adenoma (NFPA) matched by sex and age. Expression of genes involved in the regulation of bone remodeling was studied using quantitative polymerase chain reaction (qPCR). Results Of the genes involved in osteoblast and osteoclast activity, only alkaline phosphatase (ALP) mRNA was 50% downregulated in patients with acromegaly. GH excess caused increased expression of the Wnt signaling antagonists (DKK1) and agonists (WNT10B) and changes in the levels of miR involved in mesenchymal stem cell commitment to chondrocytes (miR-199a-5p) or adipocytes (miR-27-5p, miR-125b-5p, miR-34a-5p, miR-188-3p) P < 0.05; q < 0.1. Relevant compensatory mechanisms were found through the changes in miR involved in osteoblastogenesis (miR-210-5p, miR-135a-5p, miR-211, miR-23a-3p, miR-204-5p), but the expression of TWIST1 was 50% downregulated and RUNX2 was unchanged. Conclusions Acromegaly had minimal effects on tested mRNAs specific to osteoblast or osteoclast function except for downregulated ALP expression. The expressions of miR known to be involved in mesenchymal stem cell commitment and downregulated TWIST1 expression suggest acromegaly has a negative effect on osteoblastogenesis.

The effects of CD44 down-regulation on stem cell properties of head and neck cancer cell lines

2013 ◽  
Vol 42 (9) ◽  
pp. 682-690 ◽  
Author(s):  
Fahad Kidwai ◽  
Daniela E. Costea ◽  
Iain Hutchison ◽  
Ian Mackenzie
Keyword(s):  
Head And Neck Cancer ◽  
Stem Cell ◽  
Head And Neck ◽  
Cell Lines ◽  
Cancer Cell ◽  
Neck Cancer ◽  
Cancer Cell Lines ◽  
Down Regulation

scholarly journals Prevalence and genotype identification of Toxoplasma gondii in suburban rodents collected at waste disposal sites

Parasite ◽  
2019 ◽  
Vol 26 ◽  
pp. 27 ◽  
Author(s):  
Vladimir Ivovic ◽  
Sandra Potusek ◽  
Elena Buzan
Keyword(s):  
Toxoplasma Gondii ◽  
Brain Tissue ◽  
Mus Musculus ◽  
Quantitative Polymerase Chain Reaction ◽  
Rattus Rattus ◽  
Wood Mouse ◽  
Apodemus Sylvaticus ◽  
Tissue Samples ◽  
Black Rat ◽  
Two Samples

To assess the prevalence of Toxoplasma gondii infection in native and commensal rodents as indicators of environmental pollution, we analyzed brain tissue from small mammals collected on legal and illegal waste sites in the Slovenian and Croatian parts of Istria. A total of 136 animals and five species of the family Muridae were analyzed: black rat (Rattus rattus), domestic mouse (Mus musculus), wood mouse (Apodemus sylvaticus), striped field mouse (Apodemus agrarius), and yellow-necked mouse (Apodemus flavicollis). Using quantitative Polymerase Chain Reaction (qPCR), T. gondii DNA was detected in four homogenized brain tissue samples (2.94%), from all of the analyzed species, except black rat. Out of these, two samples, domestic mouse (Mus musculus) and wood mouse (Apodemus sylvaticus) had sufficient DNA for genotyping of T. gondii isolates in which we demonstrated the presence of clonal type II using RFLP PCR with four markers (SAG1, SAG2, GRA6 and GRA7). Three of four infected animals (75%) were collected on dumpsites.

scholarly journals Cancer cell line microarray as a novel screening method for identification of radioresistance biomarkers in head and neck squamous cell carcinoma

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Johannes Routila ◽  
Karri Suvila ◽  
Reidar Grénman ◽  
Ilmo Leivo ◽  
Jukka Westermarck ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Stem Cell ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Line ◽  
Cell Lines ◽  
Squamous Cell ◽  
Stem Cell Marker ◽  
Cell Marker ◽  
Hnscc Cell

Abstract Background Currently, no clinically useful biomarkers for radioresistance are available in head and neck squamous cell carcinoma (HNSCC). This study assesses the usefulness of Cell Line Microarray (CMA) method to enhance immunohistochemical screening of potential immunohistochemical biomarkers for radioresistance in HNSCC cell lines. Methods Twenty-nine HNSCC cell lines were cultured, cell pellets formalin-fixed, paraffin-embedded, and arrayed. Radioresistance features of the cell lines were combined to immunohistochemical stains for p53, NDFIP1, EGFR, stem cell marker Oct4, and PP2A inhibitor CIP2A. Results Expression of p53, EGFR or CIP2A did not indicate intrinsic radioresistance in vitro. Stem cell marker Oct4 nuclear positivity and NDFIP1 nuclear positivity was correlated with increased intrinsic radioresistance. Conclusion The usefulness of CMA in analysis of HNSCC cell lines and discovery of biomarkers is demonstrated. CMA is very well adapted to both testing of antibodies in a large panel of cell lines as well as correlating staining results with other cell line characteristics. In addition, CMA-based antibody screening proved an efficient and relatively simple method to identify potential radioresistance biomarkers in HNSCC.

Stem cell patterns in cell lines derived from head and neck squamous cell carcinoma

2007 ◽  
Vol 36 (10) ◽  
pp. 594-603 ◽  
Author(s):  
Lisa J. Harper ◽  
Kim Piper ◽  
John Common ◽  
Farida Fortune ◽  
Ian C. Mackenzie
Keyword(s):  
Squamous Cell Carcinoma ◽  
Stem Cell ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Lines ◽  
Squamous Cell ◽  
Neck Squamous Cell Carcinoma ◽  
Cell Patterns
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