Telomerase activity and p53 expression in pterygia

Shigeto Shimmura, Misaki Ishioka, Kazuomi Hanada, Jun Shimazaki, Kazuo Tsubota

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

PURPOSE. To investigate telomerase activity and p53 expression in pterygial tissue. METHODS. Pterygia tissue was obtained during excisional surgery from 35 eyes of 35 patients, and superior bulbar conjunctival tissue from the same eye was also sampled as control when possible. Fluorescence telomeric repeat amplification protocol was used to measure telomerase activity in whole pterygium samples from 9 cases and in the epithelium and stroma of pterygium from another 10 cases, p53 protein content was measured by enzyme-linked immunosorbent assay (ELISA) in tissues obtained from 7 eyes, as well as in epithelial cell suspensions collected by brush cytology in 8 eyes. Six samples were also analyzed for UV-specific mutations in the p53 gene by the single-strand conformation polymorphism technique and DNA sequencing. A conjunctival epithelial cell line was irradiated with sublethal levels of UV-B to investigate whether telomerase activity can be induced in vitro. RESULTS. In all, 63% of pterygia samples demonstrated telomerase activity, whereas all 10 paired conjunctival control samples were negative (P = 0.05, chi-square test). Of the 10 samples in which telomerase activity was measured separately in the epithelium and stroma of pterygia, 5 samples were positive in the epithelium, only 1 of which had activity in the stroma. Average telomerase activity in positive samples was 18.44 ± 8.77 U/μg protein, compared with telomerase activity measured in a carcinoma in situ patient (33.73 U/μg), and in an immortalized conjunctival epithelial cell line (50.72 ± 15.55 U/μg). Telomerase activity was not upregulated in this cell line by UV-B exposure. All 6 pterygia samples tested for p53 mutations did not reveal the UV-specific mutations in exons 5, 6, 7, or 8. No statistical significance was observed in the pterygium or conjunctiva p53 protein levels in epithelial cells collected by brush cytology, while p53 protein level was lower in pterygia when measured in whole tissue samples. CONCLUSIONS. Telomerase activity was detected in some pterygia, mostly in the epithelium. Pterygia was not associated with an increase in epithelial p53 protein content measured by ELISA.

Original languageEnglish
Pages (from-to)1364-1369
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume41
Issue number6
Publication statusPublished - 2000
Externally publishedYes

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Pterygium
Telomerase
Epithelium
Epithelial Cells
Proteins
Cell Line
Mutation
Cell Biology
Enzyme-Linked Immunosorbent Assay
Conjunctiva
p53 Genes
Carcinoma in Situ
Chi-Square Distribution
DNA Sequence Analysis
Exons
Suspensions
Fluorescence

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Telomerase activity and p53 expression in pterygia. / Shimmura, Shigeto; Ishioka, Misaki; Hanada, Kazuomi; Shimazaki, Jun; Tsubota, Kazuo.

In: Investigative Ophthalmology and Visual Science, Vol. 41, No. 6, 2000, p. 1364-1369.

Research output: Contribution to journalArticle

Shimmura, S, Ishioka, M, Hanada, K, Shimazaki, J & Tsubota, K 2000, 'Telomerase activity and p53 expression in pterygia', Investigative Ophthalmology and Visual Science, vol. 41, no. 6, pp. 1364-1369.
Shimmura, Shigeto ; Ishioka, Misaki ; Hanada, Kazuomi ; Shimazaki, Jun ; Tsubota, Kazuo. / Telomerase activity and p53 expression in pterygia. In: Investigative Ophthalmology and Visual Science. 2000 ; Vol. 41, No. 6. pp. 1364-1369.
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T1 - Telomerase activity and p53 expression in pterygia

AU - Shimmura, Shigeto

AU - Ishioka, Misaki

AU - Hanada, Kazuomi

AU - Shimazaki, Jun

AU - Tsubota, Kazuo

PY - 2000

Y1 - 2000

N2 - PURPOSE. To investigate telomerase activity and p53 expression in pterygial tissue. METHODS. Pterygia tissue was obtained during excisional surgery from 35 eyes of 35 patients, and superior bulbar conjunctival tissue from the same eye was also sampled as control when possible. Fluorescence telomeric repeat amplification protocol was used to measure telomerase activity in whole pterygium samples from 9 cases and in the epithelium and stroma of pterygium from another 10 cases, p53 protein content was measured by enzyme-linked immunosorbent assay (ELISA) in tissues obtained from 7 eyes, as well as in epithelial cell suspensions collected by brush cytology in 8 eyes. Six samples were also analyzed for UV-specific mutations in the p53 gene by the single-strand conformation polymorphism technique and DNA sequencing. A conjunctival epithelial cell line was irradiated with sublethal levels of UV-B to investigate whether telomerase activity can be induced in vitro. RESULTS. In all, 63% of pterygia samples demonstrated telomerase activity, whereas all 10 paired conjunctival control samples were negative (P = 0.05, chi-square test). Of the 10 samples in which telomerase activity was measured separately in the epithelium and stroma of pterygia, 5 samples were positive in the epithelium, only 1 of which had activity in the stroma. Average telomerase activity in positive samples was 18.44 ± 8.77 U/μg protein, compared with telomerase activity measured in a carcinoma in situ patient (33.73 U/μg), and in an immortalized conjunctival epithelial cell line (50.72 ± 15.55 U/μg). Telomerase activity was not upregulated in this cell line by UV-B exposure. All 6 pterygia samples tested for p53 mutations did not reveal the UV-specific mutations in exons 5, 6, 7, or 8. No statistical significance was observed in the pterygium or conjunctiva p53 protein levels in epithelial cells collected by brush cytology, while p53 protein level was lower in pterygia when measured in whole tissue samples. CONCLUSIONS. Telomerase activity was detected in some pterygia, mostly in the epithelium. Pterygia was not associated with an increase in epithelial p53 protein content measured by ELISA.

AB - PURPOSE. To investigate telomerase activity and p53 expression in pterygial tissue. METHODS. Pterygia tissue was obtained during excisional surgery from 35 eyes of 35 patients, and superior bulbar conjunctival tissue from the same eye was also sampled as control when possible. Fluorescence telomeric repeat amplification protocol was used to measure telomerase activity in whole pterygium samples from 9 cases and in the epithelium and stroma of pterygium from another 10 cases, p53 protein content was measured by enzyme-linked immunosorbent assay (ELISA) in tissues obtained from 7 eyes, as well as in epithelial cell suspensions collected by brush cytology in 8 eyes. Six samples were also analyzed for UV-specific mutations in the p53 gene by the single-strand conformation polymorphism technique and DNA sequencing. A conjunctival epithelial cell line was irradiated with sublethal levels of UV-B to investigate whether telomerase activity can be induced in vitro. RESULTS. In all, 63% of pterygia samples demonstrated telomerase activity, whereas all 10 paired conjunctival control samples were negative (P = 0.05, chi-square test). Of the 10 samples in which telomerase activity was measured separately in the epithelium and stroma of pterygia, 5 samples were positive in the epithelium, only 1 of which had activity in the stroma. Average telomerase activity in positive samples was 18.44 ± 8.77 U/μg protein, compared with telomerase activity measured in a carcinoma in situ patient (33.73 U/μg), and in an immortalized conjunctival epithelial cell line (50.72 ± 15.55 U/μg). Telomerase activity was not upregulated in this cell line by UV-B exposure. All 6 pterygia samples tested for p53 mutations did not reveal the UV-specific mutations in exons 5, 6, 7, or 8. No statistical significance was observed in the pterygium or conjunctiva p53 protein levels in epithelial cells collected by brush cytology, while p53 protein level was lower in pterygia when measured in whole tissue samples. CONCLUSIONS. Telomerase activity was detected in some pterygia, mostly in the epithelium. Pterygia was not associated with an increase in epithelial p53 protein content measured by ELISA.

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