Tuberc Respir Dis > Volume 69(1); 2010 > Article
Tuberculosis and Respiratory Diseases 2010;69(1):16-23.
DOI:    Published online July 1, 2010.
Induction of Autophagy by Low Dose of Cisplatin in H460 Lung Cancer Cells.
Jeong Hyun Shin, Hye Yeon Jang, Jin Soo Chung, Kyung Hwa Cho, Ki Eun Hwang, So Young Kim, Hui Jung Kim, Sam Youn Lee, Mi Kung Lee, Soon Ah Park, Sun Rock Moon, Kang Kyu Lee, Hyang Jeong Jo, Sei Hoon Yang
1Department of Internal Medicine, Wonkwang University College of Medicine, Iksan, Korea.
2Department of Thoracic Surgery, Wonkwang University College of Medicine, Iksan, Korea.
3Department of Nuclear Medicine, Wonkwang University College of Medicine, Iksan, Korea.
4Department of Therapeutic Radiology & Oncology, Wonkwang University College of Medicine, Iksan, Korea.
5Department of Pathology, Wonkwang University College of Medicine, Iksan, Korea.
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 micrometer or 20 micrometer 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 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer 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 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Key Words: Autophagy, Cisplatin, Lung Neoplasms

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