학술논문
Bufalin induced apoptosis in SCC-4 human tongue cancer cells by decreasing Bcl-2 and increasing Bax expression via the mitochondria-dependent pathway
Document Type
Report
Author
Source
Molecular Medicine Reports. December 2017, Vol. 16 Issue 6, p7959, 8 p.
Subject
Language
English
ISSN
1791-2997
Abstract
Introduction Oral cancer is one of the major causes of cancer-associated mortalities in the global human population and the total number of patients with oral cancer who are younger than [...]
The aim of the present study was to investigate the cytotoxic effects of bufalin on SCC-4 human tongue cancer cells. Cell morphological changes and viability were examined using phase contrast microscopy and flow cytometry, respectively. The results indicated that bufalin induced morphological changes and reduced total viable cells. Apoptotic cell death was analyzed by DAPI staining and DNA gel electrophoresis; the results revealed that bufalin induced cell apoptosis. Levels of reactive oxygen species (ROS), [Ca.sup.2+], nitric oxide (NO) and mitochondrial membrane potential ([DELTA][[PSI].sub.m]) were measured by flow cytometry, and bufalin was observed to increase [Ca.sup.2+] and NO production, decrease the [DELTA][[PSI].sub.m] and reduce ROS production in SCC-4 cells. In addition, western blotting was performed to detect apoptosis-associated protein expression. The results demonstrated that bufalin reduced the expression of the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2) and increased the expression of the pro-apoptotic protein, Bcl-2-associated X protein. However, bufalin treatment also increased the expression of other apoptosis-associated proteins such as apoptosis-inducing factor and endonuclease G in SCC-4 cells. Based on these findings, bufalin may induce apoptotic cell death via mitochondria-dependent pathways in human tongue cancer SCC-4 cells. Key words: bufalin, mitochondria, DNA ladder, apoptosis, SCC-4 cells
The aim of the present study was to investigate the cytotoxic effects of bufalin on SCC-4 human tongue cancer cells. Cell morphological changes and viability were examined using phase contrast microscopy and flow cytometry, respectively. The results indicated that bufalin induced morphological changes and reduced total viable cells. Apoptotic cell death was analyzed by DAPI staining and DNA gel electrophoresis; the results revealed that bufalin induced cell apoptosis. Levels of reactive oxygen species (ROS), [Ca.sup.2+], nitric oxide (NO) and mitochondrial membrane potential ([DELTA][[PSI].sub.m]) were measured by flow cytometry, and bufalin was observed to increase [Ca.sup.2+] and NO production, decrease the [DELTA][[PSI].sub.m] and reduce ROS production in SCC-4 cells. In addition, western blotting was performed to detect apoptosis-associated protein expression. The results demonstrated that bufalin reduced the expression of the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2) and increased the expression of the pro-apoptotic protein, Bcl-2-associated X protein. However, bufalin treatment also increased the expression of other apoptosis-associated proteins such as apoptosis-inducing factor and endonuclease G in SCC-4 cells. Based on these findings, bufalin may induce apoptotic cell death via mitochondria-dependent pathways in human tongue cancer SCC-4 cells. Key words: bufalin, mitochondria, DNA ladder, apoptosis, SCC-4 cells