CDDO-ME induces apoptosis in breast cancer cells via Ca2+ influx-mediated dilation of endoplasmic reticulum and c-FLIP downregulation

I.INTRODUCTION 1 II.MATERIALS AND METHODS 6 A.Chemicals and antibodies 6 B.Cell culture of various cancer cell lines 7 C.Measurement of cell viability 7 D.Western blotting 7 E.Immunocytochemistry 8 F.Establishment of the stable cell lines in the fluorescence specifically mitochondria or endoplasmic reticulum 9 G.Measurement of ROS and mitochondrial superoxide anion 9 H.Measurement of cytosolic and mitochondrial Ca² levels 9 I.shRNA-mediated knockdown of proteins 10 J.Transmission electron microscopy 10 K.Reverse transcriptionPCR analysis 11 L.Statistical analysis 12 III. RESULTS 13 1.CDDO-ME demonstrates a potent anti-cancer effect on breast cancer cells 13 2.CDDO-ME induces paraptosis-like cellular vacuolation prior to morphologies features of apoptosis in breast cancer cells 17 3.Ca2+ influx is crucial for CDDO-ME-induced vacuolation and subsequent apoptotic cell death 40 4.Cross-modulation between Ca2+ influx and ROS generation critically contributes to CDDO-ME-induced vacuolation and subsequent apoptosis 51 5.c-FLIPL downregulation plays a critical role in CDDO-ME-induced apoptotic cell death, but not in vacuolation 58 6.Higher increase in intracellular Ca2+ and ROS levels as well as c-FLIP downregulation may contribute to a more potent anti-cancer effect of CDDO-ME, compared to CDDO 69 IV.DISCUSSION 73 V.REFERENCES 79 -국문요약- 90

Author dc:creator

• 정, 수아

Contributors dc:contributor

• 최, 경숙
• 대학원 의생명과학과
• 201324121

Language dc:language

en

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000019228
000000019228

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/11827