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KMID : 0829220090330030159
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Korean Journal of Oral and Maxillofacial Pathology
2009 Volume.33 No. 3 p.159 ~ p.166
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The Effect of Cyclosporine A on Transglutaminase 2 activity and Expression of Immortalized Human Oral Keratinocytes
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You Jong-Woo
Park Gyeong-Ju
Lee Chong-Heon
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Abstract
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Cyclosporine A, immunosuppressive agent, has been applied to treat cancer cells. IHOK (Immortalized human oral keratinocyte) have been accepted as a model system for HPV-linked oral carcinogenesis. It is important to pursue the apoptosis of IHEK culture model by CsA related to the pathogenesis of oral squamous cell carcinoma. But CsA effect on immortalized human oral keratinocyte (IHOK) remains unclear. Transglutaminase 2 (TGase 2) which is expressed and activated in tissue where epithelial cells undergo apoptosis has been used as a marker for apoptotic cells. The purpose of this study were to examine the effect of CsA on the proliferation and apoptosis of cultured NHOK by TGase 2 expression. After IHOK and HN 12 cell line were treated by 0.1, 1.0 and 10ug/ml Cs A, growth curve, MTT assay for succinyl dehydrogenase activity, TGase assay for TGase 2 activity and immunoslot blotting for TGase 2 ptorein expression, and TEM features were done. MTT assay showed about 80% cell viability of HN 12, while about 50% of IHOK at 10ug/ml CsA. Growth curve showed normal S curve on control and DMSO, while IHOK was more decreased after 3 days of higher CsA treatment. Apoptosis index showed about 30% of IHOK and 15% of HN 12, respectively. TGase 2 activity and protein expression of IHOK was the highest at 10ug/ml CsA, while HN 12 was slight higher than control. TEM showed chromosome fragmentation and margination, and vacuole formation in cytoplasm of IHOK after CsA treatment. It suggested that higher cyclosporine A might induce apoptosis of IHOK as Intermediate stage cells correlated with increased TGase 2 activity and protein expression.
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KEYWORD
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Cyclosporine A, Transglutaminase 2, IHOK
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