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The Journal of Internal Korean Medicine > Volume 35(3); 2014 > Article
The Journal of Internal Korean Medicine 2014;35(3): 317-331.
桔梗에 의한 NCI-H460 인체 비소세포폐암 세포에서의 autophagy 및 apoptosis 유발 효과
홍수현1, 한민호1, 박철2, 박상은3, 홍상훈3, 최영현1,4
1동의대학교 한의과대학 생화학교실
2동의대학교 자연생활과학대학 분자생물학과
3동의대학교 한의과대학 한방내과
4동의대학교 항노화연구소 및 블루바이오소재개발센터
Induction of Autophagy and Apoptosis by the Roots of Platycodon grandiflorum on NCI-H460 Human Non-small Lung Carcinoma Cells
Su-hyun Hong1, Min-ho Han1, Cheol Park2, Sang-eun Park3, Sang-hoon Hong3, Yung-hyun Choi1,4
1Dept. of Biochemistry, College of Korean Medicine, Dong-eui University
2Dept. of Molecular Biology, College of Natural Science and Human Ecology, Dong-eui University
3Dept. of Korean Internal Medicine, College of Korean Medicine, Dong-eui University
4Anti-Aging Research Center & Blue-Bio Industry RIC, Dong-eui University
Correspondence  Su-hyun Hong ,Tel: 051-850-8644, Fax: 051-853-4036, Email: hongsh@deu.ac.kr
  Published online: September 30, 2014.
ABSTRACT
Objectives:
The root of Platycodon grandiflorum (PG) has been known to possess a range of pharmacological activities including anti-cancer, anti-inflammatory, and anti-oxidant effects. The present study was designed to investigate whether or not PG-induced cell death was connected with autophagy and apoptosis in NCI-H460 human lung cancer cells.

Methods:
Effects on the cell viability and apoptotic activity were quantified using MTT assays and flow cytometry analysis, respectively. Protein activation was measured by immunoblotting. Autophagy was measured by LC3 immunofluorescence and immunoblotting. ROS production and loss of mitochondria membrane potential (MMP) were checked with flow cytometry analysis.

Results:
Following exposure to PG, NCI-H460 cell proliferation decreased simultaneously inducing autophagic vacuoles and up-regulation of microtubule-associated protein 1 light chain 3 and beclin-1 protein expressions. Interestingly, pre-treated with autophagy inhibitors, 3-methyladenin or bafilomycin A1 further triggered reduction of cell viability. PG treatment also induced apoptosis that was related modulation of Bcl-2 family proteins, death receptors and activation of caspases. In addition, PG stimulation clearly enhanced loss of MMP and reactive oxygen species (ROS) generation.

Conclusions:
Our results suggest that PG elicited both autophagy and apoptosis by increasing loss of MMP and ROS production. PG induced-autophagy may play a cell protective role. [This research was supported by Basic Science Research Program through the NRF grant funded by the Korea government(No.2013R1A1A2065537).
Key words: Platycodong grandiflorum, Autophagy, Apoptosis, ROS, MMP
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