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

人类头颈部鳞状细胞癌中肿瘤干细胞富集表面标记 CD271 和 CD44。

  • 影响因子:4.33
  • DOI:10.1093/carcin/bgz182
  • 作者列表:"Elkashty OA","Abu Elghanam G","Su X","Liu Y","Chauvin PJ","Tran SD
  • 发表时间:2020-06-17
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.

摘要

: 头颈部鳞状细胞癌 (HNSCC) 的 5 年生存率较差,为 50%。治疗失败的一个潜在原因是癌症干细胞 (CSCs) 的存在。几种细胞标记物,特别是 CD44,已被用于分离 CSCs。然而,在 HNSCC 中分离 CSC 的纯人群仍然是一项具有挑战性的任务。最近的研究结果表明,使用 CD271 作为标记物分离出正常的口腔干细胞。因此,我们研究了联合使用 CD271 和 CD44 分离富集的 CSCs 亚群,然后在体外、体内和患者组织样本中进行表征。荧光激活细胞分选从两株人 HNSCC 细胞系中分离 CD44 +/CD271 + 和 CD44 +/CD271-。用 MTT 和球/集落形成试验测量细胞生长和自我更新。测试了对化疗 (顺铂和 5-氟尿嘧啶) 和电离辐射的治疗耐药性。QRT-PCR 检测自我更新、抗性和干性相关基因的表达。用原位免疫缺陷小鼠口腔癌模型检测体内致瘤性。最后,我们检测了患者组织样本中 CD44 +/CD271 + 的共定位。我们发现 CD271 + 细胞是人 HNSCC 细胞系和组织中 CD44 + 细胞的亚群。CD44 +/CD271 + 细胞表现出较高的细胞增殖、球/集落形成、化疗和放射抗性、 CSCs 相关基因的上调,与 CD44 +/CD271-或亲代细胞系相比,体内致瘤性。这些细胞标志物在患者中随着肿瘤分期的增加而表达增加。总之,使用 CD44 和 CD271 允许从 HNSCC 隔离 CSCs。这些富集的 CSCs 将在未来的治疗和 HNSCC 进展研究中更加相关。

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影响因子:2.65
发表时间:2020-01-29
来源期刊:BMJ open
DOI:10.1136/bmjopen-2019-031602
作者列表:["Wendland EM","Kops NL","Comerlato J","Horvath JDC","Bessel M","Sperb D","Pimenta C","de Souza FMA","Mendes Pereira GF","Falcetta FS"]

METHODS:INTRODUCTION:Human papillomavirus (HPV) is the most common sexually transmitted infection and is associated with several types of cancer. The number of cases of HPV-associated head and neck squamous cell carcinomas (HNSCCs), especially oropharyngeal carcinomas, has increased significantly in recent years despite decreased tobacco smoking rates. Currently, no data concerning the risk factors and prevalence of HPV in HNSCC patients in all regions of Brazil are available, making it difficult to promote advances in this field of public health. Therefore, our goal is to determine the impact of infection by HPV, including HPVs with different genotypes, on head and neck cancer and the risk factors associated with the development of head and neck cancer in all regions of Brazil. METHODS AND ANALYSIS:This is a case-control study that will include 622 patients and 622 controls from all regions of Brazil. A questionnaire will be applied to gather information on sociodemographic, behavioural and health factors. Oral, cervical or penile/scrotal, and anal specimens and serum samples will be collected from all participants. Formalin-fixed paraffin-embedded tissue from tumour biopsies will be analysed only in the case group. Molecular and serological analyses will be performed to evaluate the presence and role of HPV in the development of head and neck cancer. ETHICS AND DISSEMINATION:This project was approved by the research ethical committee of the proposing institution (Hospital Moinhos de Vento, number 2.852.060). Ethical approval from the collaborators is currently under evaluation and is not yet complete. The results of this study will be presented at meetings with the Brazilian Ministry of Health through technical reports and to the scientific community at national and international events, with subsequent publication of scientific articles.

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影响因子:2.60
发表时间:2020-01-21
来源期刊:Head & neck
DOI:10.1002/hed.26063
作者列表:["Soldera EB","Ortigara GB","Bonzanini LIL","Schulz RE","Danesi CC","Antoniazzi RP","Linhares Ferrazzo K"]

METHODS:BACKGROUND:Factors related to head and neck cancer and the treatment of the disease can affect quality of life. The aim of this study was to determine factors associated with the severity of impact on oral health-related quality of life (OHRQoL) in survivors of head and neck cancer using a multivariate analysis. METHODS:This cross-sectional study evaluated 90 volunteers who had completed radiotherapy at least 3 months earlier. OHRQoL was assessed using oral health impact profile (OHIP-14) and the data were analyzed using robust variance poisson regression models. RESULTS:The mean total OHIP-14 score was 23.98 ± 12.55. Patients with hyposalivation had 56% higher (worse) mean OHIP-14 total scores (CI:1.11-2.18) and patients with advanced stage tumors had 31% higher mean OHIP-14 total scores (CI:1.03-1.66) in multivariate analyses. CONCLUSION:OHRQoL of survivors of head and neck cancer experienced a negative impact following radiotherapy. The impact was associated with hyposalivation and advanced stage tumors.

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影响因子:1.62
发表时间:2020-01-22
DOI:10.4317/medoral.23335
作者列表:["Ramos-Vega V","Venegas Rojas B","Donoso Torres W"]

METHODS:BACKGROUND:To immunohistochemically evaluate the association between the presence of cancer-associated fibroblasts (CAFs) and the tumour expression of podoplanin (PDPN) in head and neck squamous cell carcinoma (HNSCC) and their association with clinicopathological variables. MATERIAL AND METHODS:A tissue microarray (TMA) with biopsy sections from patients diagnosed with HNSCC was stained with antibodies against the CAFs marker, α-smooth muscle actin (α-SMA), and PDPN. We subsequently evaluated their expression to determine the association between them and with clinicopathological variables including age, primary tumour site, TNM stage, and tumour differentiation grade. RESULTS:Positive reaction to α-SMA was observed in the tumour stroma, revealing spindle-shaped cells compatible with CAFs, which showed a high expression in 62% of cases and a significant association with laryngeal carcinomas, advanced clinical stages, and lower tumour differentiation (P ≤ 0.05). PDPN staining on tumour cells showed low expression in 72% of cases, and it was not associated with any clinicopathological variable or with the presence of CAFs. CONCLUSIONS:The presence of CAFs in the tumour stroma is related to an aggressive phenotype and could increase as the disease progresses, although based on our findings, it would have no relationship, at least directly, with the expression of PDPN.

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