Benzyl Isothiocyanate Induces Apoptotic Cell Death Through Mitochondria-dependent Pathway in Gefitinib-resistant NCI-H460 Human Lung Cancer Cells In Vitro.
BACKGROUND/AIM
Gefitinib is used to treat patients with lung cancer, but in some patients, the disease becomes gefitinib-resistant. Benzyl isothiocyanate (BITC), found in cruciferous vegetables, has shown anticancer activity in many human cancer cell lines. However, the effects of BITC on gefitinib-resistant NCI-H460 lung cancer cells in vitro have not been investigated.


MATERIALS AND METHODS
The effects of BITC on gefitinib-resistant NCI-H460 lung cancer cells were investigated in vitro. Flow cytometric assay was used for determining the total viable cell number, apoptotic cell death, the production of reactive oxygen species (ROS) and Ca2+, mitochondriaI membrane potential (Ψm) and caspase-3, -8 and -9 activities. Furthermore, 4', 6-diamidino-2-phenylindole staining was used to examine chromatin condensation in NCI-H460 and NCI-H460/G cells.


RESULTS
BITC reduced total viable cell number via the induction of apoptotic cell death, that was also confirmed by annexin V/propidium iodide double staining assay. BITC increased ROS and Ca2+ production, reduced Ψm and increased caspase-3, -8 and -9 activities in both NCI-H460 and NCI-H460/G cells. Western blotting assay also showed that BITC increased expression of cleaved caspase-3 and -9, cytochrome c, BCL2-associated X protein, endonuclease G, poly (ADP-ribose) polymerase, growth arrest and DNA-damage protein 153, caspase-7 and activating transcription factor 6 alpha, but reduced apoptosis-inducing factor and caspase-9, BH3-interacting domain death agonist, calpain 1, glucose-regulated protein 78 and inositol requiring enzyme 1 alpha in NCI-H460/G cells.


CONCLUSION
BITC-induced apoptotic cell death appears to occur via caspase- and mitochondria-dependent pathways in both cell lines.
['Huang|Yi-Ping|YP|', 'Jiang|Yi-Wen|YW|', 'Chen|Hsin-Yu|HY|', 'Hsiao|Yung-Ting|YT|', 'Peng|Shu-Fen|SF|', 'Chou|Yu-Cheng|YC|', 'Yang|Jiun-Long|JL|', 'Hsia|Te-Chun|TC|', 'Chung|Jing-Gung|JG|']
D051017:Apoptosis Regulatory Proteins D020169:Caspases D045744:Cell Line, Tumor D002470:Cell Survival D019008:Drug Resistance, Neoplasm D000077156:Gefitinib D015972:Gene Expression Regulation, Neoplastic D006801:Humans D017879:Isothiocyanates D008175:Lung Neoplasms D053078:Membrane Potential, Mitochondrial D008928:Mitochondria D011799:Quinazolines D017382:Reactive Oxygen Species D015398:Signal Transduction
Benzyl isothiocyanate mitochondria-dependent pathway gefitinib-resistant human lung cancer