Bufalin induced apoptosis in SCC-4 human tongue cancer cells by decreasing Bcl-2 and increasing Bax expression via the mitochondria-dependent pathway

Abstract

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), Ca2+, nitric oxide (NO) and mitochondrial membrane potential (ΔΨm) were measured by flow cytometry, and bufalin was observed to increase Ca2+ and NO production, decrease the ΔΨm 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.

Bufadienolides induce apoptosis and autophagy by inhibiting the AKT signaling pathway in melanoma A‑375 cells

“…First, in the mitochondrial pathway, the Bcl-2 family of proteins exerts a major role in tumorigenesis and tumor maintenance. a previous study revealed that bufadienolides may mediate apoptosis through downregulation of Bcl-2 and/or upregulation of Bax in cancer cells (24,25). in the present study, bufadienolides were identified to induce the apoptosis of a-375 cells, upregulate the expression of Bax and p53, and downregulate Bcl-2 expression.…”

Bufalin Enhances Immune Responses in Leukemic Mice Through Enhancing Phagocytosis of MacrophageIn Vivo

“…Bufalin has biological activities including inducing cancer cell apoptosis (11)(12)(13)(14)(15)(16)(17) and DNA damage in NCI-H460 human lung cancer cells (18). Recently, we also found that bufalin significantly inhibits cell adhesion, migration and invasion of gefitinib-resistant NCI-H460 cells (19) and induced apoptosis of U-2 OS human osteosarcoma cells (20) and SCC 4 human tongue cancer cells (21).…”

Divaricoside Exerts Antitumor Effects, in Part, by Modulating Mcl-1 in Human Oral Squamous Cell Carcinoma Cells

“…For example, paclitaxel suppresses cell proliferation and induces apoptosis by increasing ROS generation in canine mammary gland tumor cells [ 45 ]. Bufalin has also been shown to induce apoptosis via reduction in ROS production in tongue cancer cells [ 46 ]. Our results showed that DIV exhibits an antitumor effect by increasing ROS generation in SCC2095 cells, and this inhibition can partially be rescued by pretreatment with antioxidants.…”