Cell Cycle Arrest and Apoptosis Induction via Modulation of Mitochondrial Integrity by Bcl-2 Family Members and Caspase Dependence inDracaena cinnabari-Treated H400 Human Oral Squamous Cell Carcinoma

Abstract

Dracaena cinnabari Balf.f. is a red resin endemic to Socotra Island, Yemen. Although there have been several reports on its therapeutic properties, information on its cytotoxicity and anticancer effects is very limited. This study utilized a bioassay-guided fractionation approach to determine the cytotoxic and apoptosis-inducing effects of D. cinnabari on human oral squamous cell carcinoma (OSCC). The cytotoxic effects of D. cinnabari crude extract were observed in a panel of OSCC cell lines and were most pronounced in H400. Only fractions DCc and DCd were active on H400 cells; subfractions DCc15 and DCd16 exhibited the greatest cytotoxicity against H400 cells and D. cinnabari inhibited cells proliferation in a time-dependent manner. This was achieved primarily via apoptosis where externalization of phospholipid phosphatidylserine was observed using DAPI/Annexin V fluorescence double staining mechanism studied through mitochondrial membrane potential assay cytochrome c enzyme-linked immunosorbent and caspases activities revealed depolarization of mitochondrial membrane potential (MMP) and significant activation of caspases 9 and 3/7, concomitant with S phase arrest. Apoptotic proteins array suggested that MMP was regulated by Bcl-2 proteins family as results demonstrated an upregulation of Bax, Bad, and Bid as well as downregulation of Bcl-2. Hence, D. cinnabari has the potential to be developed as an anticancer agent.

Cryptotanshinone induces ROS-mediated apoptosis in human gastric cancer cells

“…In the extrinsic pathway, caspase-8 is activated, whereas caspase-9 is involved in the intrinsic pathway [ 21 – 23 ]. Changes in expression of pro-apoptotic protein (Bad) versus anti-apoptotic proteins (Bcl-2) activate the intrinsic apoptotic pathway [ 24 ]. It has been reported that CT can increase the expression levels of cleaved-caspase-3 and pro-apoptotic protein Bax while decrease Bcl-2 via the ROS-mitochondrial apoptotic pathway, and arrest the cell cycle at the G2/M phase in A375 melanoma cells [ 15 ].…”

Rhein Induces Cell Death in HepaRG Cells through Cell Cycle Arrest and Apoptotic Pathway

“…Moreover, the regulation and control of mitochondrial-dependent apoptotic events occur mainly through the Bcl-2 family proteins including Bcl-2, Bak, and Bax [ 24 ]. Caspases can be significantly activated by an increase in the Bax/Bcl-2 ratio, which then leads to programmed cell death through the mitochondrial-dependent apoptotic pathway [ 25 ].…”

Mequindox-Induced Kidney Toxicity Is Associated With Oxidative Stress and Apoptosis in the Mouse

“…Cleaved caspase 3 is a well-characterized cell apoptotic marker and caspase 9 is a biomarker of mitochondrial apoptosis pathway. The multimeric complex formation of cytochrome c, caspase 8, and caspase 9 activates downstream caspases, leading to apoptosis cell death ( Alabsi et al, 2016 ). Simultaneously, the Bcl-2 family proteins function as central regulators of apoptosis in mammals ( Alabsi et al, 2016 ).…”

“…The multimeric complex formation of cytochrome c, caspase 8, and caspase 9 activates downstream caspases, leading to apoptosis cell death ( Alabsi et al, 2016 ). Simultaneously, the Bcl-2 family proteins function as central regulators of apoptosis in mammals ( Alabsi et al, 2016 ). In this study, MEQ significantly increased the protein expression of cleaved caspase 3 and mRNA expression of caspase 3, suggesting that the apoptosis was induced by chronic exposure to MEQ in the kidney of mice.…”

“…In this study, the apoptosis in kidney might be resulted from the oxidative stress and mitochondrial dysfunction mediated by MEQ. Currently, there are two well-characterized caspase-activating cascades: one is initiated by the cell surface death receptor and the other is triggered by changes in mitochondrial integrity ( Alabsi et al, 2016 ). Mitochondria were reported to be a target organelle of QdNOs and extra production of ROS might be result in mitochondrial damage, following with mitochondrial apoptotic pathway ( Dai et al, 2015 ).…”