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Establishment and genetic characterization of six unique tumor cell lines as preclinical models for sinonasal squamous cell carcinoma.

García-Inclán C, López-Hernández A, Alonso-Guervós M, Allonca E, Potes S, Melón S, López F, Llorente JL, Hermsen M - Sci Rep (2014)

Bottom Line: Overexpression of p53 was observed in five, and of EGFR in four cell lines.None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus.In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

View Article: PubMed Central - PubMed

Affiliation: Dept. Otolaryngology, IUOPA, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.

ABSTRACT
Sinonasal squamous cell carcinomas (SCC) are rare tumors, etiologically related to occupational exposure to wood and leather dust. In spite of surgical and radiotherapeutic advances, the 5 year survival is still 30-50%. Therefore, alternative treatment options are needed. We report the establishment and characterization of six unique human sinonasal SCC cell lines, named SCCNC1, 2, 4, 5, 6 and 7. In vitro growth and invasion characteristics were evaluated and genetic profiles were compared to those of the original primary tumors. The population doubling times ranged from 21 to 34 hours. Cell lines SCCNC2 and 7 were highly invasive in matrigel. Five cell lines carried a high number of copy number alterations, including amplifications and homozygous deletions, while one showed only three abnormalities. Sequence analysis revealed three cell lines with TP53 mutation and none with KRAS or BRAF. Overexpression of p53 was observed in five, and of EGFR in four cell lines. None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus. In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

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P53 expression analysis by immunofluorescence on the in vitro growing cell lines (left column) and by immunohistochemistry on the corresponding primary tumors (right column), showing positivity in all cell lines except SCCNC4.
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f4: P53 expression analysis by immunofluorescence on the in vitro growing cell lines (left column) and by immunohistochemistry on the corresponding primary tumors (right column), showing positivity in all cell lines except SCCNC4.

Mentions: Sequencing of TP53 exons 5–9 revealed missense mutations in three cell lines. SCCNC1 and SCCN7 both carried a transition c.844C > T (p.R282W) in exon 8, and SCCNC5 showed a transition c.511G > A (p.E171K) in exon 5. These three cell lines also showed a strong nuclear p53 protein overexpression in the primary tumor and in the cell line. Two additional cell lines showed p53 overexpression in absence of TP53 mutation (Figure 4). EGFR gene copy number gain (all cell lines except SCCNC5) did not always associate with EGFR protein overexpression, which was observed in cell lines SCCNC1, SCCNC4, SCCNC5 and SCCNC7, but not in SCCNC2 and SCCNC6 (Figure 5). No mutations were found in KRAS exon 2 (codons 12 and 13) and BRAF exon 15 (V600E). P16 expression was absent in all cell lines, while among the primary tumors only SCCNC4 showed a weak positivity (Figure 5). HPV analysis was negative for all cell lines and primary tumors.


Establishment and genetic characterization of six unique tumor cell lines as preclinical models for sinonasal squamous cell carcinoma.

García-Inclán C, López-Hernández A, Alonso-Guervós M, Allonca E, Potes S, Melón S, López F, Llorente JL, Hermsen M - Sci Rep (2014)

P53 expression analysis by immunofluorescence on the in vitro growing cell lines (left column) and by immunohistochemistry on the corresponding primary tumors (right column), showing positivity in all cell lines except SCCNC4.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4017217&req=5

f4: P53 expression analysis by immunofluorescence on the in vitro growing cell lines (left column) and by immunohistochemistry on the corresponding primary tumors (right column), showing positivity in all cell lines except SCCNC4.
Mentions: Sequencing of TP53 exons 5–9 revealed missense mutations in three cell lines. SCCNC1 and SCCN7 both carried a transition c.844C > T (p.R282W) in exon 8, and SCCNC5 showed a transition c.511G > A (p.E171K) in exon 5. These three cell lines also showed a strong nuclear p53 protein overexpression in the primary tumor and in the cell line. Two additional cell lines showed p53 overexpression in absence of TP53 mutation (Figure 4). EGFR gene copy number gain (all cell lines except SCCNC5) did not always associate with EGFR protein overexpression, which was observed in cell lines SCCNC1, SCCNC4, SCCNC5 and SCCNC7, but not in SCCNC2 and SCCNC6 (Figure 5). No mutations were found in KRAS exon 2 (codons 12 and 13) and BRAF exon 15 (V600E). P16 expression was absent in all cell lines, while among the primary tumors only SCCNC4 showed a weak positivity (Figure 5). HPV analysis was negative for all cell lines and primary tumors.

Bottom Line: Overexpression of p53 was observed in five, and of EGFR in four cell lines.None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus.In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

View Article: PubMed Central - PubMed

Affiliation: Dept. Otolaryngology, IUOPA, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.

ABSTRACT
Sinonasal squamous cell carcinomas (SCC) are rare tumors, etiologically related to occupational exposure to wood and leather dust. In spite of surgical and radiotherapeutic advances, the 5 year survival is still 30-50%. Therefore, alternative treatment options are needed. We report the establishment and characterization of six unique human sinonasal SCC cell lines, named SCCNC1, 2, 4, 5, 6 and 7. In vitro growth and invasion characteristics were evaluated and genetic profiles were compared to those of the original primary tumors. The population doubling times ranged from 21 to 34 hours. Cell lines SCCNC2 and 7 were highly invasive in matrigel. Five cell lines carried a high number of copy number alterations, including amplifications and homozygous deletions, while one showed only three abnormalities. Sequence analysis revealed three cell lines with TP53 mutation and none with KRAS or BRAF. Overexpression of p53 was observed in five, and of EGFR in four cell lines. None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus. In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

Show MeSH
Related in: MedlinePlus