Sulforaphane‐induced apoptosis in human leukemia HL‐60 cells through extrinsic and intrinsic signal pathways and altering associated genes expression assayed by cDNA microarray

Abstract

Sulforaphane (SFN), one of the isothiocyanates, is a biologically active compound extracted from cruciferous vegetables, and has been shown to induce cytotoxic effects on many human cancer cells including human leukemia cells. However, the exact molecular mechanism and altered gene expression associated with apoptosis is unclear. In this study, we investigated SFN-induced cytotoxic effects and whether or not they went through cell-cycle arrest and induction of apoptosis and further examined molecular mechanism and altered gene expression in human leukemia HL-60 cells. Cell viability, cell-cycle distribution, sub-G1 (apoptosis), reactive oxygen species (ROS) and Ca production, levels of mitochondrial membrane potential (ΔΨ ), and caspase-3, -8, and -9 activities were assayed by flow cytometry. Apoptosis-associated proteins levels and gene expressions were examined by Western blotting and cDNA microarray assays, respectively. Results indicated that SFN decreased viable cells, induced G2/M phase arrest and apoptosis based on sub-G1 phase development. Furthermore, SFN increased ROS and Ca production and decreased the levels of ΔΨ and activated caspase-3, -8, and -9 activities in HL-60 cells. SFN significantly upregulated the expression of BAX, Bid, Fas, Fas-L, caspase-8, Endo G, AIF, and cytochrome c, and inhibited the antiapoptotic proteins such as Bcl-x and XIAP, that is associated with apoptosis. We also used cDNA microarray to confirm several gene expressions such as caspase -8, -3, -4, -6, and -7 that are affected by SFN. Those results indicated that SFN induced apoptosis in HL-60 cells via Fas- and mitochondria-dependent pathways. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 311-328, 2017.

Fisetin Induces Apoptosis of HSC3 Human Oral Cancer Cells Through Endoplasmic Reticulum Stress and Dysfunction of Mitochondria-mediated Signaling Pathways

“…Caspases, cysteinyl aspartate-specific proteases, are involved in anticancer drug-induced cancer cell apoptosis, and can be activated during apoptosis in a self-amplifying cascade (49,50). Our results revealed that fisetin significantly increased caspase-3, -8 and -9 expression (Figure 5), showing the involve¬ment of caspases in apoptosis of HSC3 cells.…”

Sulforaphane displays the growth inhibition, cytotoxicity and enhancement of retinoic acid-induced superoxide-generating activity in human monoblastic U937 cells

“…Moreover, in human being, sulforaphane displays the multiple interesting biological functions: such as anti-cancer effects (Li et al, 2011;Amin and Shankar, 2015;Dandawate et al, 2016;Leone et al, 2017;Jiang et al, 2018;Sita et al, 2018), antioxidant functions (Negrette-Guzman et al, 2013), anti-Helicobacter pylori activity (Yanaka, 2017), photo aging prevention (Sikdar et al, 2016), prevention of obesity (Martins et al, 2018), and general health benefits (Vanduchova et al, 2019). On the other hand, although numerous reports concerning to the chemopreventive and chemotherapeutic properities of solid tumors are available, there is little information on the properities of sulforaphane in leukemia cells (Choi et al, 2008;Fimognari et al, 2008;Suppipat and Lacorazza, 2014;Fimognari et al, 2014;Shih et al, 2016;Shang et al, 2017;Brown et al, 2017;Koolivand et al, 2018). Unfortunately, the understanding of the effects of sulforaphane on human leukemia cells with mechanisms is incompletely explored till now.…”

ROS-mediated activation of AMPK plays a critical role in sulforaphane-induced apoptosis and mitotic arrest in AGS human gastric cancer cells

“…This phytochemical has a strong antioxidant potential and has proven to be an important cancer-preventive candidate, exhibiting high activity that inhibits the proliferation of cancer cells and induces apoptosis in many cancers (Gupta et al 2014;Sestili and Fimognari 2015). Although inhibition of the proliferation of cancer cells by sulforaphane in some cell lines has been reported to be associated with G1 arrest in the cell cycle (Shen et al 2006;Bryant et al 2010;Wang et al 2012), many studies have shown that it induces G2/M arrest associated with apoptosis in multiple cancer types (Singh et al 2004;Matsui et al 2007;Chu et al 2009;Suppipat et al 2012;Wang et al 2015;Byun et al 2016;Shang et al 2017). The effects of sulforaphane on cell cycle progression have also been reported to be specific for mitotic arrest, but not the G2 phase, which is associated with destruction of microtubule polymerization (Jackson and Singletary 2004;Mi et al 2008;Xiao et al 2012).…”

“…The effects of sulforaphane on cell cycle progression have also been reported to be specific for mitotic arrest, but not the G2 phase, which is associated with destruction of microtubule polymerization (Jackson and Singletary 2004;Mi et al 2008;Xiao et al 2012). In addition, it has been shown in various previous studies that ROS act as key regulators in the induction of apoptosis by sulforaphane (Singh et al 2004;Wang et al 2015;Byun et al 2016;Shang et al 2017). Further, recent results from Mondal et al (2016) demonstrate that induction of apoptosis and inhibition of migration of human gastric cancer cells by sulforaphane are associated with increased ROS production and blocking of several intracellular signaling pathways.…”