Effects of hTERT antisense oligodeoxynucleotide on cell apoptosis and expression of hTERT and bcl-2 mRNA in keloid fibroblasts

Y. Huang, L.-X. Lin, Q.-X. Bi, P. Wang, X.-M. Wang, J. Liu, Y.-T. Wang

Department of Burn and Plastic Surgery, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China. hyong915@sina.com

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• Pharmacology

OBJECTIVE: This study was purposed to investigate the effects of hTERT antisense oligodeoxynucleotide (ASODN) on cell apoptosis and expression of hTERT and bcl-2 mRNA in keloid fibroblasts and to explore its anti-keloid effect.

MATERIALS AND METHODS: Primary cultures of dermal fibroblasts derived from 12 keloid samples were established, strains of fibroblasts at passages 3 to 4 were used in this study. After treated by hTERT ASODN the proliferation of the fibroblasts was measured by cell count and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay method, the apoptosis was analyzed by flow cytometry (FCM), and the expression of hTERT and bcl-2 mRNA were observed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The data was analyzed by statistical software (SPSS11.5).

RESULTS: The results showed that after sealing hTERT gene with ASODN for 72 h, the fibroblasts growth was repressed and the ability of proliferation decreased as the fibroblasts were treated with 1.0 mol/L ASODN for 72 h, the fibroblasts apoptosis was induced and the expression of hTERT and bcl-2 mRNA was lower than that of controlled group. The result was significantly different between control group and treatment group and was related to the treatment time of ASODN (p<0.01), but the difference was no significant when compared 1.0 μmol/L SODN group with untreated group (p>0.05).

CONCLUSIONS: As a negative modutory factor, hTERT-ASODN can suppress growth and proliferation of keloid fibroblasts. Decreasing the telomerase activity of keloid fibroblasts may be one of the most important mechanisms. That hTERT-ASODN inhibited telomerase activity in keloid fibroblasts is an important pathway that may play a key role in the anti- keloid therapy.

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