Establishment and Characterization of a Novel Head and Neck Squamous Cell Carcinoma Cell Line USC-HN1
© Liebertz et al; licensee BioMed Central Ltd. 2010
Received: 23 January 2010
Accepted: 22 February 2010
Published: 22 February 2010
Head and neck squamous cell carcinoma (HNSCC) is an aggressive and lethal malignancy. Publically available cell lines are mostly of lingual origin, or have not been carefully characterized. Detailed characterization of novel HNSCC cell lines is needed in order to provide researchers a concrete keystone on which to build their investigations.
The USC-HN1 cell line was established from a primary maxillary HNSCC biopsy explant in tissue culture. The immortalized cells were then further characterized by heterotransplantation in Nude mice; immunohistochemical staining for relevant HNSCC biomarkers; flow cytometry for surface markers; cytogenetic karyotypic analysis; human papillomavirus and Epstein-Barr virus screening; qRT-PCR for oncogene and cytokine analysis; investigation of activated, cleaved Notch1 levels; and detailed 35,000 gene microarray analysis.
Characterization experiments confirmed the human HNSCC origin of USC-HN1, including a phenotype similar to the original tumor. Viral screening revealed no HPV or EBV infection, while western blotting displayed significant upregulation of activated, cleaved Notch1.
USC-HN1, a novel immortalized cell line has been derived from a maxillary HNSCC. Characterization studies have shown that the cell line is of HNSCC origin and displays many of the same markers previously reported in the literature. USC-HN1 is available for public research and will further the investigation of HNSCC and the development of new therapeutic modalities.
Current research has delineated many generalized and specific markers to characterize HNSCC cell lines. Histologically, HNSCC is a squamous epithelial carcinoma with variable degrees of keratinization. Well-differentiated cell lines may display keratin pearls, whereas poorly differentiated, anaplastic cell lines may have little-to-no keratin production. HNSCC is typically characterized by a malignant phenotype including large, pleomorphic nuclei and large or multiple nucleoli; cytoplasmic vacuolation with abundant cytoplasm; intercellular bridging; and high numbers of mitotic figures, both typical and atypical. Beside these morphologic features, surface and intracellular markers are also used to identify the cell line lineage. Along with traditional markers such as FABP5, epidermal growth factor receptor, E-cadherin, CD74, and CD24, newly published biomarkers for the staining of HNSCC primary tumor biopsies include IL13Rα2, CD44v6, and the stem cell marker CD133 . The population of cancer stem cells (CSC) within the tumor biopsy represented by CD44+CD133+ cells has been shown to have a high incidence of metastasis and invasion. These cells, however, have been found to be a rare subset of cells (2-10%) in the overall tumor cell population [7, 8]. Karyotypic analysis of HNSCC typically reveals an aberrant chromosome set with various deletions, translocations, and double chromosomes, including a deletion of the small arm of chromosome 3 found in many epithelial cancers . Human papilloma virus (HPV), specifically HPV subtype-16 is associated with HNSCC in about 30% of cases and is most commonly associated with tumors arising from the oropharynx. HPV-positive HNSCC is now considered a distinct biomodel from HPV-negative HNSCC, occurring in patients without the usual history of alcohol and/or tobacco use . Epstein-Barr virus (EBV) is a commonly-associated Herpes virus linked with nasopharyngeal carcinoma not found in HNSCC. Bergmann et al.  have reported extensive evidence of the immunomodulatory effects of HNSCC, including the local and regional suppression of the immune system by interleukins, TGF-β, and other cytokines. Oncogenes such as c-myc, and c-Kit and mutations in tumor suppressor genes p53 and Rb are also characteristically found in HNSCC [4, 11–13]. Lastly, it has been reported that Notch1, an embryonically-associated receptor for inhibition of differentiation, may play a role in the oncogenesis of multiple types of cancers including leukemias, lung, melanoma, breast, and neurological tumors  but its role in HNSCC has not been studied extensively to date.
In this report, we now describe the establishment and characterization of a unique HNSCC cell line designated USC-HN1. The cell line was derived from an invasive primary right superior alveolar ridge squamous cell carcinoma in a nonsmoking patient (Stage IVa, T4aN0M0, based on the American Joint Commission on Cancer Staging, 6th Ed.) and has been found to recapitulate the phenotype of the original tumor biopsy. Heterotransplantation and cytogenetic studies demonstrate its oncogenic derivation and monoclonality, respectively. This cell line has been made available for others in the scientific community through the American Tissue-type Cell Collection (ATCC, http://www.atcc.org) and represents an important model for further studies of HNSCC.
Cell Lines and Cells
HeLa, HUT102, Raji, FaDu, and SW579 cell lines were obtained from the ATCC. All cell lines were maintained in complete medium in humidified incubators at 37°C with 5% CO2. IRB approval from the USC Keck School of Medicine (HS-09-00048) has been obtained for the collection and use of HNSCC tumor biopsies. Tumor biopsies were surgically resected and placed into 50 mL collection tubes containing approximately 30 mL of RPMI-1640 medium with 20% FCS, 1% Antibiotic-Antimycotic Solution (Mediatech, Inc., Manassas, VA), 10 ug/ml Ciprofloxacin-HCl (Mediatech, Inc.), and 10 ug/ml Gentamicin Sulfate (Irvine Scientific, Santa Ana, CA). The tubes were immediately put on ice and transported to the laboratory for tissue dissociation.
Establishment of Cell Line USC-HN1
Tumor biopsies were mechanically dissociated into small tissue fragments using scissors and forceps. Fragments were then further dissociated enzymatically in 20 ml of RPMI-1640 medium containing 10% FCS, and 0.2 micron membrane filtered 0.01% hyaluronidase, 0.1% collagenase, and 0.01% DNase (Sigma Chemical Co., St. Louis, MO) for 40 min in a 37°C water bath with intermittent mixing. After digestion, the cells and fragments were washed once in complete medium (RPMI-1640 medium containing 20% FCS, 1% Antibiotic-Antimycotic Solution, 10 ug/ml ciprofloxacin HCl, and 10 μg/ml Gentamicin Sulfate), treated for 30 seconds in 30 ml sterile filtered RBC lysis buffer (8.3 g NH4Cl, 1 g KHCO3, and 0.037 g EDTA/L dH2O), washed again in complete medium, and seeded in two T-75 flasks. After 2-3 days of incubation in a humidified 5% CO2 37°C incubator, the fragments were repeatedly pipetted with a 2 ml glass pipette for further dissociation. After centrifugation, the cells and residual small fragments were resuspended in freezing medium (RPMI-1640 medium with 30% FCS, 1% Penicillin/Streptomycin, and 10% DMSO), aliquoted into four-six 1.8 ml cryotubes (Nunc, Denmark), and placed in liquid nitrogen for long-term storage.
Cells and fragments that adhered to the T-75 flasks were grown in complete medium for 2 weeks before being trypsinized and passaged weekly to new flasks. When the malignant cells were seen to grow, the normal fibroblastic cells were removed by differential trypsinization to enrich the malignant cell population. Finally, malignant cells were cloned in petri dishes using cloning rings to isolate a pure population of HNSCC cells to establish the cell line. The doubling time was determined by cell count measurements at 24 hr intervals for one week from cells in culture after trypsinization. After establishment of the cell line, interval screening for Mycoplasma was performed using the MycoAlert Mycoplasma Detection Kit (Lonza, Rockland, ME).
Heterotransplantation in Nude mice
Six-week-old female Nude mice were purchased from Harlan Sprague Dawley (Indianapolis, IN). Institutional Animal Care and Use Committee-approved protocols and institutional guidelines for the proper humane care and use of animals in research were followed. Mice (n = 5) were injected s.c. in the flank with a 0.2 mL inoculum of 5 × 106 viable USC-HN1 cells. Three weeks after implantation, tumors were removed and the tissue was either flash frozen in liquid nitrogen or fixed in 10% neutral buffered formalin overnight at room temperature for paraffin-embedded procedures.
For immunohistochemistry (IHC) studies, cytospin preparations of USC-HN1 cells and tissue sections of USC-HN1 tumors grown in Nude mice were used and compared to stained, fixed slides of the original tumor. USC-HN1 cells grown for 24 hr directly on sterile printed 25 × 75 mm glass slides (Bellco Glass, Inc., Vineland, NJ) were fixed sequentially with 2% parafolmaldehyde (Polysciences, Inc., Warrington, PA) for 10 min at room temperature and acetone for 5 min at -20°C. For tissue sections, excised heterotransplanted USC-HN1 tumors from Nude mice were fixed overnight in 10% neutral buffered formalin and embedded in paraffin blocks. For cell culture and tissue morphology studies, Wright-Giemsa and hematoxilin & eosin stains were used, respectively on air-dried cytospin preparations. In addition, USC-HN1 cytospin preparations and 5 micron tissue sections were stained with monoclonal antibodies against human CD44 (clone DF1485 Dako Corp., Carpinteria, CA), E-cadherin (clone 4A2C7 Invitrogen, Carlsbad, CA), epidermal growth factor receptor (EGFr) (clone E30 Biogenex, San Ramon, CA), keratin (clone AE1/AE-3 Covance, Berkeley, CA), p53 (clone 1801 CalBiochem, San Diego, CA), and Rb (clone RbG3-245 BD Biosciences, San Diego, CA). Observation, evaluation and image acquisition were made using Leica DM2500 microscope (Leica Microsystems, http://www.leica-microsystems.com) connected to an automated, digital SPOT RTke camera and SPOT Advanced Software (SPOT Diagnostic Instrument Inc., http://www.diaginc.com). Images were further resized and brightened for publication using Adobe Photoshop software (Adobe, http://www.adobe.com).
Single cell suspensions (1 × 106 cells in 100 μl) in FACS buffer (1% FCS in PBS) were stained with FITC, PE, PerCPCy5.5, and APC conjugated antibodies. For intracellular staining, cell surface staining was performed first, followed by buffer fixation/permeabilization (eBioscience, San Diego, CA) and intracellular staining. Antibodies used were: CD24 (ML5), CD74 (M-B741), E-cadherin (36/E-cadherin) (BD Biosciences); IL-13Rα (B-D13), c-Kit (104D2) (Santa Cruz Biotechnology, Santa Cruz, CA); CD44v6 (VFF-7) (Abcam, Cambridge, MA); CD133 (TMP4) (eBioscience); FABP5 (311215) (R&D Systems, Minneapolis, MN); and anti-human Epidermal Growth Factor Receptor (CTL-R2) (Cancer Therapeutics Laboratories, Inc., Los Angeles, CA). Staining with isotype controls antibodies (eBioscience) was performed in parallel, and all samples were done in duplicate. Samples were run on a FACS Calibur flow cytometer (BD) and data acquisition and analysis were performed using Cell Quest Pro software (BD) at the USC Flow Cytometry core facility.
Karyotype analysis was performed by the Division of Anatomic Pathology, City of Hope (Duarte, CA) using cultured USC-HN1 cells.
Polymerase chain reaction (PCR) viral screen
Genomic DNA was isolated from USC-HN1, HeLa, HUT102, Raji, FaDu, and SW579 cells using TRIreagent (Sigma) per manufacturer's instructions. For PCR, 50-100 ng of DNA was amplified with specified primers (300 nM final concentration) using REDTaq ReadyMix PCR Master Mix (Sigma) in a 25 μl reaction and run on an iCycler (BioRad, Hercules, CA). HPV infectivity was screened using previously reported consensus primers MY09/MY11 (expected product ~450 bp) and GP5+/GP6+ (expected product ~150 bp) [15, 16]. Consensus primers for the EBV gene EBNA2 (expected product ~600 bp) was used to screen for the presence of EBV as previously reported .
Cytokine and oncogene analysis by qRT-PCR
Cytokine and Oncogene Primers for qRT-PCR
5' - GCTCGACGCTAGGATCTGAC - 5'
5' - CAGGTAGCTGCTGGGCTC - 3'
5' - GTTGGTCCTTCTCGGTCCTT - 3'
5' - CAAAGCAGAAGGCAACTTGA - 3'
5' - CTCCTCCTCGTCGCAGTAGA - 3'
5' - GCTGCTTAGACGCTGGATTT - 3'
5' - GCCCACGCGGACTATTAAGT - 3'
5' - CTGGGATTTTCTCTGCGTTC - 3'
5' - CACACAGGATGGCTTGAAGA - 3'
5' - AGGGCAGAATCATCACGAAG - 3'
5' - CTCCAGATCTTTGCTTGCAT - 3'
5' - CTGTGGCGTGTTCTCTGCT - 3'
5' - TTCAAATGAGATTGTGGGAAAATTGCT - 3'
5' - AGATCATCTCTGCCTGAGTATCTT - 3'
5' - GCAGAAGTTGGCATGGTAGC - 3'
5' - CCCTGGACACCAACTATTGC - 3'
5' - CTCCATTGCTGAGACGTCAA - 3'
5' - CGACGAAGAGTACTACGCCA - 3'
5' - GGAGATTCGTAGCTGGATGC - 3'
5' - GAGCTCGCCAGTGAAATGAT - 3'
5' - AGCGAGTGTCCTTCTCATGG - 3'
5' - CAGCCTCACAGAGCAGAAGA - 3'
5' - CATTTGTGGTTGGGTCAGG - 3'
5' - AGTGAGGAACAAGCCAGAGC - 3'
5' - AGCACTCCTTGGCAAAACTG - 3'
5' - CAAGAGCCAGGAAGAAACCA - 3'
5' - GCCACCCTGATGTCTCAGTT - 3'
5' - GTGGAGCAGGTGAAGAATGC - 3'
5' - CTCTGCTCCTCCTGTTCGAC - 3'
5' - TTAAAAGCAGCCCTGGTGAC - 3'
Western blot for activated Notch1
For western blots, 50 μg of protein from sonicated whole cell lysates were fractionated in a 10% Tris-glycine polyacrylamide gel, electrotransferred to PDVF membranes, and probed overnight with primary antibody for activated Notch1 (clone Val1744) (Cell Signaling, Danvers, MA). Blots were stripped and reprobed for GAPDH (clone FL-335) (Santa Cruz Biotech), to normalize the amount of sample loaded. Horseradish peroxidase-conjugated secondary antibodies (Caltag, Burlingame, CA) were then applied, followed by signal detection using Immobilon Western Chemiluminescent HRP Substrate (Millipore, Billerica, MA).
Microarray gene expression profiling
Total RNA from USC-HN1 was collected and analyzed by microarray as previously described . Briefly, 5 μg of total RNA was reverse-transcribed using 5'-amino-modified primers with amino-allyl-dUTP. cDNA synthesized from USC-HN1 cells was labeled with Cy5 dye, and cDNA from universal RNA (uRNA) labelled with Cy3 dye. Labeled and combined cDNA probes were denatured, mixed in SlideHyb #1 hybridization buffer (Ambion, Austin, TX), and placed onto microarray slides. Thirty five thousand oligonucleotide arrays were hybridized at 42°C in MAUI hybridization system (BioMicro Systems) for overnight, washed with 1 × SSC with 0.05% SDS and 0.1 × SSC buffers, and then were quickly spin-dried. Microarray slides were scanned on a GenePix 4000B scanner (Axon Instruments, Inc., Foster City, CA) with a 5 μm resolution. Scanned raw images were analyzed and data files were generated with GenePix Pro 5.1 (Axon) software. For analysis, data files were uploaded into mAdb (microarray database), and analyzed by the software tools provided by the Center for Information Technology (CIT), NIH. A standard global normalization approach was used for each experiment. All of the extracted data was normalized using a 50th percentile (median) normalization method. Statistical analyses and t-test were performed to identify differentially expressed genes.
Case report of patient NR with Invasive Right Maxillary SCC
Establishment of the USC-HN1 cell line
Heterotransplantation into Nude mice
Immunophenotype of USC-HN1 in cell culture and in situ
USC-HN1 characterization by flow cytometry
Analysis of USC-HN1 cytokines and surface markers by FACS.
Cytogenetic analysis of the USC-HN1 cell line was performed in collaboration with the City of Hope (Duarte, CA) (Figure 7). All mitotic cells analyzed from the USC-HN1 cell line were clonally abnormal. The complex near-triploid clone was characterized by a modal number of chromosomes of 71 and by deletions involving chromosomes X, 3, 4, 6, and 7, additional material of unknown origin on chromosomes 8, 13, 17, and 19, a derivative chromosome 6 resulting from an unbalanced translocation with the long arm of chromosome 14, gains of chromosomes X, 7, 11, 15, and 19, losses of chromosomes 9, 14, 18, 21, and 22, as well as gain of a large marker chromosome and a small suspected ring chromosome.
PCR Viral screen for HPV and EBV
Cytokine and oncogenes expression
Oncogene and cytokine analysis of HNSCC cell lines by qRT-PCR.
Average Fold Δ
Average Fold Δ
Microarray gene expression profiling
Selected up-regulated genes identified in USC-HN1 cells by microarray analysis also present in HNSCC tumor biopsies.*
GeneBank Access ID
Gene Symbol & Annotation
Log2 Fold Difference**
MIF, macrophage migration inhibitory factor
HLA-A, major histocompatibility complex, class I A
IGFBP2, insulin-like growth factor binding protein 2
CD24, CD24 antigen
Cell Growth, Maintenance/Cell cycle Regulation
KRT18, keratin 18
KRT8, keratin 8
CFL1, cofilin 1
PFN1, profilin 1
KRT19, keratin 19
FTL, ferritin, light polypeptide
CCT4, chaperonin subunit 4
Translation and Protein Synthesis
RPLP0, ribosomal protein LP0
RPL18, ribosomal protein L18
RPL10, ribosomal protein L10
RPS18, ribosomal protein S18
EIF4G2, translation initiation factor 4 gamma 2
RPL37A, ribosomal protein L37A
RPS 3A, ribosomal protein S3A
RPL21, ribosomal protein L21
EIF4A2, eukaryotic translation initiation factor 4A
EEF1D, eukaryotic translation elongation factor 1D
PKM2, pyruvate kinase, muscle
PPIA, peptidylprolyl isomerase A (cyclophilin A)
LDHB, lactate dehydrogenase B
HSPB1, heat shock 27 kDa protein
PPM1G, protein phosphatase 1G
ATP5G3, ATP synthase H+ transporting subunit
ATP5H, ATP synthase H+ transporting subunit
PSMB5, proteasome subunit, beta 5
GPX1, glutathione peroxidase 1
PPP2CA, protein phosphatase 2 catalytic subunit
ATP1B3, ATP synthase Na+/K+ transporting, beta 3
FABP5, fat acid binding protein 5
Ion Binding Proteins
S100A11, S100 calcium binding protein A11
S100A14, S100 calcium binding protein A14
TMSB10, thymosin, beta 10
ENO1, enolase 1
UBC, ubiquitin C
PRDX1, peroxiredoxin 1
NME2, non-metastatic cells 2 protein
CD164, CD164 antigen
We describe the clinical presentation of an invasive well-differentiated maxillary HNSCC and the establishment of the USC-HN1 cell line from the primary tumor biopsy. Of note, patient NR was a non-smoker and did not receive preoperative radiotherapy or chemotherapy. Her tumor was consistent with a diagnosis of primary HNSCC on presentation by tissue histology and immunostaining for HNSCC markers. The USC-HN1 cell line was established in culture approximately 4 weeks after seeding by isolation from co-existing fibroblastic monolayers which grew from the explanted cultures. USC-HN1 shares many characteristics with the primary tumor and has a phenotype typical of an advanced HNSCC. The doubling time of the cell line is extremely rapid (18 hr) compared to other human tumor cell lines from HNSCC  or those from other tumor types.
To demonstrate its malignant potential, the cell line was successfully heterotransplanted into Nude mice where it was found to produce invasive tumors. In comparison to the original tumor, the heterotransplant most prominently shows a higher nuclear to cytoplasmic ratio, markedly increased mitotic activity, prominent necrosis and less conspicuous intercellular bridging. Marked keratinization is also absent. The difference between the original tumor histology and the subsequently developed cell line's heterotransplant is likely due to the outgrowth of a more aggressive, less differentiated cell sub-population during establishment of the cell line in culture. In addition, sections of the heterotransplanted tumors stained for HNSCC markers were mostly consistent with previously reported primary tumor biopsies including strong EGFr, p53, and Rb staining, as is seen in many epithelial malignancies . Furthermore, fixed sections of the original tumor biopsy and cytospin cell preparations were also stained for these biomarkers. Noted differences of the cytospin preparations relative to the heterotransplanted tumors were a decrease in keratin and EFGr staining and an increase in E-cadherin and CD44 positivity. The moderate staining of keratin found in the heterotransplant tumor sections is similar with the original tumor, described as a keratinizing squamous cell carcinoma. The light staining of E-cadherin in the heterotransplanted tumor may indicate the malignant potential of the cells as they begin to undergo epithelial-to-mesenchymal transition and consequently lose cell-cell adhesive and interactive molecules [12, 19]. When the malignant cells are placed in a murine host, they are subjected to a myriad of microenvironmental factors different from that of cell culture medium or a human host. The loss of CD44 in the heterotransplanted tumor sections may be explained by the presence of factors that increase the differentiation of USC-HN1 in situ in contrast to its stem cell-like phenotype that predominates in cell culture or in vivo .
Analysis of the flow cytometry data supports a comparable picture to the IHC in which almost all USC-HN1 cells were positive for FABP5 and half were E-cadherin positive (97.5% and 49.7%, respectively). Furthermore, USC-HN1 showed a significant increase in EGFr staining as is to be expected in an epithelial malignancy . Interestingly, stem cell markers were not uniformly expressed in the sample of cultured HNSCC although CD24 and CD74 showed increased staining unlike CD133. USC-HN1 is grown in a serum-containing medium, which may irreversibly differentiate any stem cells in the population and therefore alter the surface marker expression in the remaining pluripotent cells . Conversely, these cells may represent only a subpopulation of tumor stem cells found previously to display various combinations of CD24, CD74, and CD133 . Our results did not show a significant amount of surface staining for the recently published markers IL-13Rα or CD44v6. However, these published studies were performed on primary tumor biopsies and not on established cell lines . The karyotype analysis of USC-HN1 did show a highly abnormal chromosome content consistent with other known HNSCC cell lines and solid tumors. Aneusomy is a recurrent finding in cytogenetically abnormal head and neck tumor specimens . Deletions are the most common structural rearrangement observed, particularly involving the short arms of chromosomes 3 and 8, as well as the short arm regions of the acrocentric chromosomes. Many structural breakpoints in HNSCC involve centromeric or pericentromeric regions of the chromosomes, often resulting in isochromosome formation or derivative whole-arm translocations involving the acrocentric chromosomes [Personal communication with Dr. Joyce L. Murata-Collins, Division of Anatomic Pathology, City of Hope, Duarte, CA]. Most notably, the partial deletion of chromosome 3 and the near-triploid karyotype of the clonal population are consistent with the HNSCC-derived nature of this cell line . Viral oncogene investigation detected no infection with either HPV or EBV. In this regard, 80-90% of nasopharyngeal carcinomas have been shown to be infected with EBV but very few (<1%) of HNSCC are positive. An extensive literature report revealed only two cases of EBV+ HNSCC and both cases were co-infected with HPV [21, 22]. The EBV-negative characteristic of USC-HN1 further supports the origin of the tumor as a squamous and not a nasopharyngeal carcinoma. By contrast, 30% of HNSCC are infected with a subtype of HPV . USC-HN1 represents a unique HPV-negative HNSCC cell line in that it was derived from a patient who did not smoke tobacco or drink alcohol and does not have a family history of head and neck malignancies.
Further characterization of the USC-HN1 cell line revealed a similar pattern of cytokine and chemokine expression compared to the pharyngeal carcinoma FaDu, a widely used HNSCC cell line. As previously reported HNSCC are highly immunomodulatory and alter their tumor microenvironment by the production of various cytokines . The production of VEGFc has been correlated with increased metastatic potential in HNSCC , and USC-HN1 has shown a statistically increased production of VEGFc compared with FaDu. Importantly, with the exception of c-myc, the USC-HN1 cell line reveals statistically equivalent expression of proto-oncogenes and tumor-suppressor genes as FaDu and a cytokine expression profile different from FaDu, which will offer researchers yet another biomodel to study the microenvironment of HNSCC. Finally, the increased levels of activated, cleaved Notch1 found in USC-HN1 are indicative of its malignant potential, and although published reports imply various levels of Notch1 activity among HNSCC cell lines , it may serve as a future avenue for a new therapeutic approach since multiple trials of Notch1 inhibitors are in progress in patients with other tumor types .
Microarray analysis of USC-HN1 revealed a pattern of gene expression similar to HNSCC tumor samples previously reported . In addition, specific up-regulated genes including CD24, E-cadherin, FABP5, keratins, heat shock protein (27 kDa), and others identified by microarray analysis were also identified by flow cytometry and immunostaining. These results support the origin of the cell line and further confirm the pathology as HNSCC.
In conclusion, we have established a novel HPV-negative HNSCC cell line from a non-smoking elderly patient. USC-HN1, one of only a few cell lines derived from the upper alveolar ridge, recapitulates the primary tumor's malignant behavior. It also displays surface and intracytoplasmic biomarkers consistent with HNSCC. This cell line will provide researchers with a well-delineated model to investigate the oncogenesis of HNSCC and may provide a source of material to develop new therapeutic reagents capable of treating these deep-seated and highly resistant tumors.
The authors wish to acknowledge the expert technical assistance of the City of Hope Cytogenetic Core Facility (Duarte, CA) in performing the cytogenetic studies on the USC-HN1 cell line, Ms. Lillian Young for her help with the IHC studies, Dr. Clive Taylor for his assistance with microphotography, Dr. Dixon Gray for his help with the flow cytometry studies, and Mr. James Pang for performing the animal investigations. This work was supported in part by a grant from the American Type Tissue Collection and from funds provided by Cancer Therapeutics Laboratories, Los Angeles, CA.
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