Cell Death Effects Induced by Sulforaphane and Allyl Isothiocyanate on P-Glycoprotein Positive and Negative Variants in L1210 Cells

Abstract

Variants of L1210 leukemia cells-namely, parental P-glycoprotein-negative S cells and R and T cells expressing P-glycoprotein, due to selection with vincristine and transfection with the human p-glycoprotein gene, respectively-were used. The responses of these cell variants to two naturally occurring isothiocyanates-sulforaphane (SFN, from cruciferous vegetables) and allyl isothiocyanate (AITC, from mustard, radish, horseradish and wasabi)-were studied. We obtained conflicting results for the cell death effects induced by isothiocyanates, as measured by i. cell counting, which showed inhibited proliferation, and ii. cell metabolic activity via an MTS assay, which showed an increased MTS signal. These results indicated the hyperactivation of cell metabolism induced by treatment with isothiocyanates. In more detailed study, we found that, depending on the cell variants and the isothiocyanate used in treatment, apoptosis and necrosis (detected by annexin-V cells and propidium iodide staining), as well as autophagy (detected with monodansylcadaverine), were involved in cell death. We also determined the cell levels/expression of Bcl-2 and Bax as representative anti- and pro-apoptotic proteins of the Bcl-2 family, the cell levels/expression of members of the canonical and noncanonical NF-κB pathways, and the cell levels of 16 and 18 kDa fragments of LC3B protein as markers of autophagy.

Chloroquine commonly induces hormetic dose responses

“…In contrast, the studies of Cudazzo et al (2019) and Martins et al (2013) suggested that chloroquine enhanced the uptake of crystal violet dye in a biphasic concentration manner in HaCaT cells but that this biphasic response did not occur using the NRU assay. A similar divergent finding as seen in 2019) was recently made by Kontar et al (2020). They reported that sulforaphane induced biphasic dose responses in three types of L1210 leukemia cells with the MTS cell viability assay, while, at the same time, showing a dose-dependent decrease in cell number, in contrast to the expected increase in cell number.…”

“…They reported that sulforaphane induced biphasic dose responses in three types of L1210 leukemia cells with the MTS cell viability assay, while, at the same time, showing a dose-dependent decrease in cell number, in contrast to the expected increase in cell number. Possible reasons for this endpoint decoupling/divergence was suggested by Kontar et al (2020). A similar type of focus for the divergent findings with the NRC assay with chloroquine remains to be made.…”

“…In response to the SARS epidemic, Saverino et al (2003) suggested that chloroquine may be of clinical value in the management of SARS. This suggestion was based on its effectiveness as an antimalarial agent and its anti-viral activity against HIV type 1 ( Tsai et al, 1990;Pardridge et al, 1998;Saverino et al, 2001) hepatitis virus (Kouroumalis andKoskinas, 1986), herpes simplex virus type 1 (Singh et al, 1996), and HCoV-229e (Blau and Holmes, 2001). Based on this literature, Keyaerts et al (2004) provided a dose response inhibitory effect of chloroquine on the yield of SARS-COV when grown in Vero E6 cells.…”

Sulphoraphane Affinity-Based Chromatography for the Purification of Myrosinase from Lepidium sativum Seeds

“…The antitumor effects of isothiocyanates result from multilevel modification of various biological regulatory pathways. Their efficacy may vary in different neoplastic cells and may induce arrest of cell cycling [11], induction of apoptosis [11], or autophagy [12], as well as many other processes that limit disease progression.…”

Comparative Effects of Sulforaphane and Allyl Isothiocyanate on 3T3-L1 Adipogenesis

“…The logP, or the water and n -octanol partition coefficient, is 0.23 ± 0.39 for sulforaphane and 1.77 ± 0.30 for allyl isothiocyanate [ 20 ], indicating that sulforaphane is more hydrophilic than allyl isothiocyanate. This is likely due to differences in the physicochemical properties of the side chains, excluding the ITC group [ 7 , 8 ], and indicates that the nonspecific passage of sulforaphane into the cell is less favorable than that of allyl isothiocyanate; instead, sulforaphane may bind specifically to certain proteins expressed on the cell surface.…”