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KMID : 0383820100690010016
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Tuberculosis and Respiratory Diseases
2010 Volume.69 No. 1 p.16 ~ p.23
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Induction of Autophagy by Low Dose of Cisplatin in H460 Lung Cancer Cells
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Shin Jeong-Hyun
Park Soon-Ah
Lee Sam-Youn
Jo Hyang-Jeong
Yang Sei-Hoon
Kim So-Young
Jang Hye-Yeon
Hwang Jong-Yun
Kim Hui-Jung
Moon Sung-Rock
Lee Mi-Kyung
Lee Kang Kyu
Chung Jin-Soo
Cho Kyung-Hwa
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Abstract
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Background: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells.
Methods: H460 cells were incubated with RPMI 1640 and treated in 5 μM or 20 μM cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy.
Results: Lung cancer cells treated with 5 μM cisplatin-treated were less sensitive to cell death than 20 μM cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 μM was not detected, even though it was discovered at 20 μM. Poly (ADP-ribose) polymerase cleavages were not detected in 5 μM within 24 hours. Massive vacuolization in the cytoplasm of 5 μM treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 μM treated cells, but was not detected in 20 μM treated cells. The expression of GFP-LC3 were increased in 5 μM treated cells in a time-dependent manner.
Conclusion: The induction of autophagy occurred in 5 μM dose of cisplatin-treated lung cancer cells.
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KEYWORD
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Autophagy, Cisplatin, Lung Neoplasms
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