본 연구는 topoisomerase inhibitors가 배양 각막 상피세포에서 세포고사를 유도하 는지를 조사기 위해 시행하였다. Topoisomerase inhibitors'{l camptothecin과 etoposide 를 제조사의 추천농도로 1-2 일 동안 배양하여 MTT assay를 이용하여 세포독성을 검 정하여 농도틀 정하고 세포고사를 확인하였다 세포고사의 형태적인 특징은 Hoechst 33342 staining, Annexin V - FITC/PI staining, DePsipher assay and CytoDEA TH staining을 이용하여 확인하였고 DNA fragmentation은 TUNEL파 agarose gel 전기 영동으로 확인하였다. Camptothecin 과 etoposide는 농도 의존성으로 세포고사룹 유또 하였는데 각막상피세포에서는 저l 조사의 추천농도보다 낮은 농도에서 세포고사플 유 도하여 다른 세포보다 민감한 것으로 나타났다- 이러한 결파로 볼 때 topOlsomerase l띠1ibitor는 각막상피세포에서 홀륭한 양성 대조군으로 활용될 수 있을 것으로 사료된다.

This study was performed to examine the apoptosis induced in human comeal epithelial cells maintained in tissue culture by topoisomerase inhibitors. Topoisomerase inhibitors, camptothecin and etoposide at manufacturer-recommended concentration were added to the comeal epithelial cells for 24 or 48 hours. Cell viability was measured using MTT assay. The cells were then assessed for the level of apoptosis Morphologic features of apoptosis were demonstrated using Hoechst 33342 staining, Annexin V - FITC/PI staining, DePsipher assay and CytoDEA TH staining. DNA fragmentation was determined by means of an in situ cell detection procedure (TUNEL) or by electrophoresis on agarose gels. Camptothecin and etoposide induced apoptosis in HCE cells in a dose dependent manner. Camptothecin and etoposide induced apoptosis at lower than manufacturer recommended concentration. Camptothecin and etoposide elicited typical apoptotic morphologic changes. Topoisomerase inhibitors are more sensitive in apoptotic induction of HCE cells than other cells. These results indicate that topoisomerase inhibitors are able to play a role in good positive control of apoptotic inducers

• 김재민(건양대학교 안경광학과) | Jai-Min Kim