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

Other Title
악성 유방암 세포에서 CDDO-ME 처리 시 Ca2+ 유입에 매개된 소포체 팽창과 c-FLIP 감소를 통한 세포자살 유도
Authors
정, 수아
Department
대학원 의생명과학과
Degree
Master (2015)
Table Of Contents
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
Appears in Collections:
Theses > Graduate School of Biomedical Sciences > Master
AJOU Authors
정, 수아
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