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http://dx.doi.org/10.5352/JLS.2015.25.1.53

Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937  

Ha, Jun Young (Department of Life Science & Environmental Biochemistry, Pusan National University)
Kim, Mi Kyeong (Department of Life Science & Environmental Biochemistry, Pusan National University)
Lee, Jun Young (Department of Life Science & Environmental Biochemistry, Pusan National University)
Choi, Eun Bi (Department of Life Science & Environmental Biochemistry, Pusan National University)
Hong, Chang Oh (Department of Life Science & Environmental Biochemistry, Pusan National University)
Lee, Byong Won (Department of Functional Crop Nat'l Institute of Crop Science, RDA)
Bae, Chang Hwan (Wild Life Genetic Resource Center, National Institute of Biological Resources, Environment Research Complex)
Kim, Keun Ki (Department of Life Science & Environmental Biochemistry, Pusan National University)
Publication Information
Journal of Life Science / v.25, no.1, 2015 , pp. 53-61 More about this Journal
Abstract
In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.
Keywords
Apoptosis; Perilla frutescens; photosensitizer; porphyrin; U937;
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