Separation of nasopharyngeal epithelial cells from carcinoma cells on 3D scaffold platforms

Nestin, a class VI intermediate filament protein, is known to be expressed in various types of human neoplasms, including breast cancer, and is associated with their progression. However, its expression and role in canine mammary tumors remain unknown. We analyzed nestin expression in canine mammary tumors using in situ hybridization and immunohistochemistry. We also investigated its role in a canine mammary carcinoma cell line using RNA interference. Nestin expression was not observed in luminal epithelial cells of any of the 62 cases of benign mammary lesions examined, although myoepithelial cells showed its expression in most cases. In 16/50 (32%) primary mammary carcinomas and 6/15 (40%) metastases of mammary carcinomas, cytoplasmic nestin expression was detected in luminal epithelial cells. In luminal cells of primary mammary carcinomas, its expression was positively related to several pathological parameters that indicate high-grade malignancy, including histological grading ( P < .01), vascular/lymphatic invasion ( P < .01), Ki-67 index ( P < .01), and metastasis ( P < .05). Immunohistochemistry revealed that nestin expression was related to vimentin expression in mammary carcinomas ( P < .01). This relationship was confirmed using reverse transcription-quantitative polymerase chain reaction using 9 cell lines derived from canine mammary carcinoma ( P < .01). Finally, nestin knockdown in canine mammary carcinoma cells using small interfering RNA inhibited cell proliferation and migration based on WST-8, Boyden chamber, and cell-tracking assays. These findings suggest that nestin may at least partially mediate these behaviors of canine mammary carcinoma cells.

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Rat carcinoma cells in long-term, serum-free culture provide a continuing supply of collagenase

Bioscience Reports ◽

10.1007/bf01119628 ◽

1985 ◽

Vol 5 (12) ◽

pp. 1071-1077 ◽

Cited By ~ 11

Author(s):

Geoffrey A. Stevenson ◽

J. Guy Lyons ◽

David A. Cameron ◽

Robert L. O'Grady

Keyword(s):

Epithelial Cells ◽

Bovine Serum Albumin ◽

Serum Albumin ◽

Mammary Carcinoma ◽

Bovine Serum ◽

Defined Medium ◽

Carcinoma Cells ◽

Serum Free ◽

Vertebrate Collagenase

Neoplastic, epithelial cells derived from a spontaneously-arising rat mammary carcinoma have been cultured in a defined medium, in the absence of serum, continuously, for over 2 years. The medium is a mixture of Ham's F12 and Dulbecco's Modified Eagle's media supplemented with insulin, transferrin and bovine serum albumin. The cells have retained their potential to produce tumours and, in culture, a true vertebrate collagenase. This system provides a continuing supply of vertebrate collagenase through the application of recently developed methods.

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Molecular and cytogenetic changes involved in the immortalization of nasopharyngeal epithelial cells by telomerase

International Journal of Cancer ◽

10.1002/ijc.22032 ◽

2006 ◽

Vol 119 (7) ◽

pp. 1567-1576 ◽

Cited By ~ 81

Author(s):

H.M. Li ◽

C. Man ◽

Y. Jin ◽

W. Deng ◽

Y.L. Yip ◽

...

Keyword(s):

Epithelial Cells ◽

Cytogenetic Changes ◽

Nasopharyngeal Epithelial

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Enzymatic Modulation of the Cell Surface in Malignant Transformation of Normal Human Mammary Epithelial Cells and in Conversion of Mammary Carcinoma Cells

Pathobiology ◽

10.1159/000162953 ◽

1979 ◽

Vol 47 (5) ◽

pp. 332-350

Author(s):

Anwar A. Hakim

Keyword(s):

Epithelial Cells ◽

Cell Surface ◽

Malignant Transformation ◽

Mammary Carcinoma ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Carcinoma Cells ◽

Human Mammary Epithelial ◽

Mammary Carcinoma Cells ◽

Normal Human

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Transgelin, a p53 and PTEN-Upregulated Gene, Inhibits the Cell Proliferation and Invasion of Human Bladder Carcinoma Cells in Vitro and in Vivo

International Journal of Molecular Sciences ◽

10.3390/ijms20194946 ◽

2019 ◽

Vol 20 (19) ◽

pp. 4946 ◽

Cited By ~ 6

Author(s):

Tsui ◽

Lin ◽

Chang ◽

Hou ◽

Chen ◽

...

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Bladder Carcinoma ◽

Carcinoma Cells ◽

Human Bladder ◽

Ectopic Overexpression ◽

Proliferation And Invasion ◽

Bladder Carcinoma Cells

Transgelin (TAGLN/SM22-α) is a regulator of the actin cytoskeleton, affecting the survival, migration, and apoptosis of various cancer cells divergently; however, the roles of TAGLN in bladder carcinoma cells remain inconclusive. We compared expressions of TAGLN in human bladder carcinoma cells to the normal human bladder tissues to determine the potential biological functions and regulatory mechanisms of TAGLN in bladder carcinoma cells. Results of RT-qPCR and immunoblot assays indicated that TAGLN expressions were higher in bladder smooth muscle cells, fibroblast cells, and normal epithelial cells than in carcinoma cells (RT-4, HT1376, TSGH-8301, and T24) in vitro. Besides, the results of RT-qPCR revealed that TAGLN expressions were higher in normal tissues than the paired tumor tissues. In vitro, TAGLN knockdown enhanced cell proliferation and invasion, while overexpression of TAGLN had the inverse effects in bladder carcinoma cells. Meanwhile, ectopic overexpression of TAGLN attenuated tumorigenesis in vivo. Immunofluorescence and immunoblot assays showed that TAGLN was predominantly in the cytosol and colocalized with F-actin. Ectopic overexpression of either p53 or PTEN induced TAGLN expression, while p53 knockdown downregulated TAGLN expression in bladder carcinoma cells. Our results indicate that TAGLN is a p53 and PTEN-upregulated gene, expressing higher levels in normal bladder epithelial cells than carcinoma cells. Further, TAGLN inhibited cell proliferation and invasion in vitro and blocked tumorigenesis in vivo. Collectively, it can be concluded that TAGLN is an antitumor gene in the human bladder.

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Epstein–Barr Virus Infection of Pseudostratified Nasopharyngeal Epithelium Disrupts Epithelial Integrity

Cancers ◽

10.3390/cancers12092722 ◽

2020 ◽

Vol 12 (9) ◽

pp. 2722

Author(s):

Fenggang Yu ◽

Yanan Lu ◽

Yingying Li ◽

Yuji Uchio ◽

Utomo Andi Pangnguriseng ◽

...

Keyword(s):

Epithelial Cells ◽

Epstein Barr Virus ◽

Basal Layer ◽

Infection Model ◽

Hybridization Technique ◽

Epstein Barr Virus Infection ◽

Barr Virus ◽

Ebv Infection ◽

Epstein Barr ◽

Nasopharyngeal Epithelial

Epstein–Barr virus (EBV) is a human oncogenic virus that causes several types of tumor, such as Burkitt’s lymphoma and nasopharyngeal carcinoma (NPC). NPC tumor cells are clonal expansions of latently EBV-infected epithelial cells. However, the mechanisms by which EBV transforms the nasopharyngeal epithelium is hampered, because of the lack of good in vitro model to pursue oncogenic process. Our primary nasopharyngeal epithelial cell cultures developed pseudostratified epithelium at the air-liquid interface, which was susceptible to EBV infection. Using the highly sensitive RNA in situ hybridization technique, we detected viral infection in diverse cell types, including ciliated cells, goblet cells, and basal cells. EBV-encoded small RNA-positive cells were more frequently detected in the suprabasal layer than in the basal layer. We established the most physiologically relevant EBV infection model of nasopharyngeal epithelial cells. This model will advance our understanding of EBV pathogenesis in the development of NPC.

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