KGN(난소과립세포)에서 BCL2L10 단백질의 세포사멸 유도 기능 연구
BCL2L10 Protein Induces Apoptosis in KGN-human Granulosa Cells
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초록
BCL-2 family 단백질들은 세포사멸 신호전달 체계에서 중추적인 역할을 수행하는 것으로 알려져 있으며, BCL2L10 단백질은 그 중 하나로 세포의 사멸과 생존을 조절하는 것으로 알려져 있다. 특이하게도 BCL2L10 단백질은 세포 또는 조직 특이적으로 서로 상반되는 친 세포사멸 또는 항 세포사멸 효과가 각각 보고되어 있다. 현재까지 난소세포에서의 BCL2L10의 발현 여부 및 기능은 알려져 있지 않다. 따라서 본 연구에서 인간 난소 과립세포주인 KGN 세포에서의 BCL2L10 단백질의 발현 여부를 확인한 결과, 상당한 양의 단백질이 발현함을 확인할 수 있었으며, 또한 세포사멸 효과를 확인하기 위해서 BCL2L10 단백질을 dose-dependent하게 과발현시킨 후 세포의 생존에 미치는 영향을 분석한 결과, BCL2L10은 KGN 세포에서 과발현 시 세포사멸을 유도함을 관찰하였다. BCL2L10 단백질을 과발현 시 Caspase 9와 3를 활성화 하였으며, 면역염색법을 통해서 BCL2L10 단백질이 미토콘드리아에 위치하는 것을 확인하였다. 또한 BCL2L10 단백질의 과발현에 의해 미토콘드리아에서 cytochrome c가 세포질로 분비되는 것을 확인하였다. 이상의 결과로서 본 연구는 BCL2L10의 과발현이 KGN 세포에서 세포사멸을 유도하고, 또한 미토콘드리아에 위치하여 세포질로 cytochrome c를 분비하여 Caspase 9와 3을 활성화 시키는 메커니즘으로 세포사멸을 유도함을 확인하였다. 이러한 연구결과는 BCL2L10 단백질이 인간 난소과립세포의 생존과 사멸을 조절하는 인자임을 최초로 규명한 것으로서, 추후 난소에서의 BCL2L10 단백질의 생리적인 기능 및 신호 조절 연구의 기반 데이터로서 그 의의가 있으며 활용될 수 있다.
Abstract
BCL-2 family essential proteins to play a pivotal role to perform in apoptosis signaling pathways and essential proteins for the regulation of cell death. BCL2L10 protein is a member of BCL-2 family and it regulates both anti-apoptotic and pro-apoptotic function of specific tissue or cell line. BCL2L10 of function and expression is not reported in ovary cell lines. In this study we reported that BCL2L10 were significant expression of KGN cell line. Ectopic expression of BCL2L10 induced cell death, and its cells killing effect was blocked by pan-caspase inhibitor of the Z-VAD-fmk. Ectopic expression of BCL2L10 protein led to the activation of caspase 9 and caspase 3, suggesting apoptotic cell death, and confocal microscopic analyses showed that BCL2L10 was partially localized in mitochondria. Thus, we provide a novel function of BCL2L10 in KGN cells, which was involved in the intrinsic cell death pathway.
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