Loperamide enhances bortezomib-mediated cell death in colon cancer cells

I. INTRODUCTION 1 II. MATERIALS AND METHODS 8 A. Chemicals and antibodies 8 B. Cell culture of various cancer cell lines and normal cells 8 C. Measurement of cell viability 9 D. Western blotting 9 E. Immunocytochemistry 10 F. Establishment of the stable cell lines in the fluorescence specifically endoplasmic reticulum 10 G. Measurement of mitochondrial superoxide anion 10 H. Measurement of cytosolic and mitochondrial Ca² levels 11 I. Small interfering RNAs 11 J. shRNA-mediated knockdown of proteins 12 K. Aggresome staining 12 L. Flow cytometry for the analysis of DNA contents 13 M. Clonogenic cell survival assay 13 III. RESULTS 14 1. Synergistic cell death induced by loperamide plus bortezomib is neither apoptosis not autophagic cell death 14 2. Intracellular Ca2+ influx is involved in the combined treatment-induced vacuolation 27 3. Combined treatment with loperamide and bortezomib induces ER stress, and CHOP is critically involved in the cell death by loperamide plus bortezomib 33 4. ROS generation is important for the cell death induced by loperamide plus bortezomib 42 5. Noxa is also critical factor for loperamide plus bortezomib-induced cell death 46 6. Disruption of aggresome formation may be involved in the cell death induced by loperamide and bortezomib 50 7. Combined treatment with loperamide and proteasome inhibitors does not induce cell death in normal colon cells 56 IV. DISCUSSION 60 V. REFERENCES 64 - 국문요약 - 77

Author dc:creator

• 심, 민준

Contributors dc:contributor

• 최, 경숙
• 대학원 의생명과학과
• 201224473

Language dc:language

en

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000019150
000000019150

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/11833