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영양분이 결핍된 H460 세포주에서 자가포식이 세포사멸에 미치는 영향
장혜연 ( Hye Yeon Jang ),조향정 ( Hyang Jeong Jo ),황기은 ( Ki Eun Hwhang ),김소영 ( So Young Kim ),이강규 ( Kang Kyoo Lee ),문성록 ( Sun Rock Moon ),신정현 ( Jeong Hyun Shin ),조경화 ( Kyung Hwa Cho ),이미경 ( Mi Kung Lee ),이삼 대한결핵 및 호흡기학회 2010 Tuberculosis and Respiratory Diseases Vol.69 No.2
Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.
폐암세포주에서 저용량 시스플라틴에 의해 유도된 자가포식
신정현 ( Jeong Hyun Shin ),장혜연 ( Hye Yeon Jang ),정진수 ( Jin Soo Chung ),조경화 ( Kyung Hwa Cho ),황기은 ( Ki Eun Hwang ),김소영 ( So Young Kim ),김휘정 ( Hui Jung Kim ),이삼윤 ( Sam Youn Lee ),이미경 ( Mi Kung Lee ),박순아 ( 대한결핵 및 호흡기학회 2010 Tuberculosis and Respiratory Diseases Vol.69 No.1
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.