난소암에서 Paclitaxel의 내성에 관여하는 Clusterin
Clusterin confers paclitaxel resistance in ovarian cancer
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목적: 난소암에서 clusterin의 과발현이 paclitaxel의 내성과 유의한 연관성이 있는지 알아본다. 연구 방법: Expression profiling 분석을 통하여 선별된 clusterin에 대하여 clusteirn mRNA 발현정도와 paclitaxel의 IC50를 비교하여 clusterin의 발현과 paclitaxel 내성의 연관성 여부를 조사한 후 clusterin 유전자와 clusterin siRNA을 paclitaxel에 감수성이 높고 clusteirn 발현이 적은 SKOV3 세포주와, paclitaxel에 내성을 가지고 있으며 clusterin 발현이 높은 PEOH 세포주에 각각 transfection시켜 난소암에서 cluterin이 항암제 내성, apoptosis 및 세포주기 변화에 미치는 영향을 XTT assay, cell death ELISA, flow cytometry를 이용하여 확인한다. 결과: Clusterin mRNA와 clusterin 단백의 발현정도는 paclitaxel 약제내성과 유의한 연관성을 보였다 (P<0.001). 또한 SKOV3 세포주에 clusterin 유전자를 transfection한 경우는 paclitaxel 치료 시 transfection하지 않은 대조군에 비하여 apoptosis의 현저하 저하를 보였으며 이와는 반대로 PEON세포주에 clusterin siRNA를 transfection한 경우는 대조군에 비하여 현저한 paclitaxel 내성 저하를 보였고 (P<0.05) S기에서 G2/M기로의 세포주기의 분포 이동을 보였다. 결론: 이 같은 실험결과로 clusterin은 난소암 세포에서 apoptosis와 세포주기의 변화를 통해 paclitaxel 약제내성에 관여하는 것으로 추정된다.
Abstract
Objective: To evaluated whether clusterin over-expression is significantly correlated with paclitaxel resistance in ovarian cancer cell lines. Methods: Clusterin was validated by performing expression profiling analysis and subsequently, the correlation between clusterin mRNA expression levels and the IC50 of paclitaxel was tested. Transfection of clusterin was performed on SKOV3, which expressed paclitaxel- sensitivity and low level of clusterin, and transfection of clusterin siRNA on PEOH, which expressed paclitaxel-resistance and high level of clusterin, to evaluate their effect on chemo-sensitivity, apoptosis, and cell cycle by XTT assay, cell death ELISA, and flow cytometry, respectively. Results: Clusterin mRNA and protein expression levels were significantly correlated with paclitaxel resistance (P<0.001). Transfection of cluterin on SKOV3 significantly decreased apoptosis and increased paclitaxel resistance. And transfection of clusterin siRNA on PEOH significantly increased paclitaxel-sensitivity (P<0.05), and shifted cells from S to G2/M phase of the cell cycle after paclitaxel treatment. Conclusion: These findings suggested that clusterin overexpression confers paclitaxel-resistance by the modulation of the apoptotic pathway and cell cycle progression in ovarian cancer cells.
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