Cearoin Induces Autophagy, ERK Activation and Apoptosis via ROS Generation in SH-SY5Y Neuroblastoma Cells.
Neuroblastomas are the most common solid extracranial tumors in childhood. We investigated the anticancer effect of cearoin isolated fromDalbergia odoriferain human neuroblastoma SH-SY5Y cells. SH-SY5Y cells were treated with various doses of cearoin. The viability was measured by MTT assay. DCFDA fluorescence assay and Griess assay were used for the measurement of intracellular reactive oxygen species (ROS) and nitric oxide (NO), respectively. Western blot analysis was performed to clarify the molecular pathway involved. Cearoin induced cell death in a dose-dependent manner. Cearoin increased the phosporylation of ERK, the conversion of LC3B-I to LC3B-II, decrease in Bcl2 expression, the activation of caspase-3, and the cleavage of PARP, indicating the induction of autophagy and apoptosis. Furthermore, cearoin treatment increased the production of ROS and NO. Co-treatment with the antioxidantN-acetylcysteine completely abolished cearoin-mediated autophagy, ERK activation and apoptosis, suggesting the critical role of ROS in cearoin-induced anticancer effects. Moreover, co-treatment with ERK inhibitor PD98059 partially reversed cearoin-induced cell death, indicating the involvement of ERK in cearoin anticancer effects. These data reveal that cearoin induces autophagy, ERK activation and apoptosis in neuroblastoma SH-SY5Y cells, which is mediated primarily by ROS generation, suggesting its therapeutic application for the treatment of neuroblastomas.
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. tonkingbastola@gmail.com.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. tonkingbastola@gmail.com.
Tonking
T
Bastola
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. anrb@ybu.edu.cn.|Laboratory of Natural Resources of Changbai Mountain & Functional Molecules Yanbian University, Ministry of Education, Yanji 133002, Jilin, China. anrb@ybu.edu.cn.
Ren-Bo
RB
An
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. yckim@wku.ac.kr.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. yckim@wku.ac.kr.
Youn-Chul
YC
Kim
Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. medicdog@wku.ac.kr.|Department of Meridian & Acupoint, College of Korean Medicine, Wonkwang University, Iksan 570-749, Korea. medicdog@wku.ac.kr.
Jaehyo
J
Kim
Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan 570-749, Korea. jwseo@wku.ac.kr.|Hanbang Body-Fluid Research Center, Wonkwang University, Iksan 570-749, Korea. jwseo@wku.ac.kr.
Jungwon
J
Seo
D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D035782:Dalbergia / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*
D000972:Antineoplastic Agents, Phytogenic; D000975:Antioxidants; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D048049:Extracellular Signal-Regulated MAP Kinases
ERK; apoptosis; autophagy; cearoin; neuroblastoma; reactive oxygen species