Curcumin induces apoptosis through FAS and FADD, in caspase-3-dependent and -independent pathways in the N18 mouse-rat hybrid retina ganglion cells.
Curcumin, a naturally occurring yellow pigment isolated from turmeric, is a well known antioxidant with broad spectrum of anti-tumor activities against many human cancer cells. In this study, curcumin-induced cytotoxic effect of mouse-rat hybrid retina ganglion cells (N18) were investigated. For determining cell viability, the trypan blue exclusion and flow cytometric analysis were used. The curcumin-caused cell cycle arrest in N18 cells was examined by flow cytometry. Curcumin affect on the production of reactive oxygen species and Ca2+ and on the decrease of the level of mitochondria membrane potential (DeltaPsim) were also examined by flow cytometry. Curcumin-induced apoptosis was determined by DAPI staining and Western blotting was used for examining the apoptotic signaling proteins. Cell cycle analysis showed that G2/M phase arrest and sub-G1 occurs in N18 cells following 48 h exposure to curcumin. Curcumin also caused a marked increase in apoptosis, as characterized by DNA fragmentation (sub-G1 phase formation) and DAPI staining, and dysfunction of mitochondria, which was associated with the activation of caspase-8, -9 and -3. Curcumin also promoted the levels of Fas and FADD, Bax, cytochrome c release, but decreased the levels of Bcl-2 causing changes of DeltaPsim. Curcumin also induced endoplasmic reticulum stress in N18 cells which was based on the changes of GADD153 and GRP78 and caused Ca2+ release. Curcumin induced apoptosis through the intrinsic pathway and caspase-3-dependent and -independent pathways in N18 cells.
Lu
Hsu-Feng
HF
Department of Clinical Pathology, Cheng Hsin Rehabilitation Medical Center, Taipei, R.O.C.
Lai
Kuang-Chi
KC
Hsu
Shu-Chun
SC
Lin
Hui-Ju
HJ
Yang
Mei-Due
MD
Chen
Yuan-Liang
YL
Fan
Ming-Jen
MJ
Yang
Jai-Sing
JS
Cheng
Pi-Yun
PY
Kuo
Chao-Lin
CL
Chung
Jing-Gung
JG
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003474:Curcumin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism*; D006822:Hybrid Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D015398:Signal Transduction / Q000187:drug effects*; D013997:Time Factors; D019014:fas Receptor / Q000378:metabolism*
D000972:Antineoplastic Agents, Phytogenic; D000091342:Endoplasmic Reticulum Chaperone BiP; D053200:Fas-Associated Death Domain Protein; C000717247:HSPA5 protein, human; C000717248:Hspa5 protein, mouse; D017382:Reactive Oxygen Species; D019014:fas Receptor; D053148:Caspase 3; D003474:Curcumin; D002118:Calcium