Increasing the TRPM2 Channel Expression in Human Neuroblastoma SH-SY5Y Cells Augments the Susceptibility to ROS-Induced Cell Death

Abstract

Human neuroblastoma SH-SY5Y cells are a widely-used human neuronal cell model in the study of neurodegeneration. A recent study shows that, 1-methyl-4-phenylpyridine ion (MPP), which selectively causes dopaminergic neuronal death leading to Parkinson’s disease-like symptoms, can reduce SH-SY5Y cell viability by inducing H2O2 generation and subsequent TRPM2 channel activation. MPP-induced cell death is enhanced by increasing the TRPM2 expression. By contrast, increasing the TRPM2 expression has also been reported to support SH-SY5Y cell survival after exposure to H2O2, leading to the suggestion of a protective role for the TRPM2 channel. To clarify the role of reactive oxygen species (ROS)-induced TRPM2 channel activation in SH-SY5Y cells, we generated a stable SH-SY5Y cell line overexpressing the human TRPM2 channel and examined cell death and cell viability after exposure to H2O2 in the wild-type and TRPM2-overexpressing SH-SY5Y cells. Exposure to H2O2 resulted in concentration-dependent cell death and reduction in cell viability in both cell types. TRPM2 overexpression remarkably augmented H2O2-induced cell death and reduction in cell viability. Furthermore, H2O2-induced cell death in both the wild-type and TRPM2-overexpressing cells was prevented by 2-APB, a TRPM2 inhibitor, and also by PJ34 and DPQ, poly(ADP-ribose) polymerase (PARP) inhibitors. Collectively, our results show that increasing the TRPM2 expression renders SH-SY5Y cells to be more susceptible to ROS-induced cell death and reinforce the notion that the TRPM2 channel plays a critical role in conferring ROS-induced cell death. It is anticipated that SH-SY5Y cells can be useful for better understanding the molecular and signaling mechanisms for ROS-induced TRPM2-mediated neurodegeneration in the pathogenesis of neurodegenerative diseases.

Resveratrol attenuates hypoxia-induced neuronal cell death, inflammation and mitochondrial oxidative stress by modulation of TRPM2 channel

“…2. Accumulating evidence indicates that SH-SY5Y cell has a PARP-1 activation 28,30 . The PARP-1 expression, TRPM2 mRNA expression, IL-1β and TNF-α release levels were higher in the SH-SY5Y as compared to the control and RSV (p ≤ 0.001).…”

TRPM2 channel-mediated cell death: An important mechanism linking oxidative stress-inducing pathological factors to associated pathological conditions

“…As discussed above, the activity of nuclear PARP, particularly PARP-1, is crucial for the induction of ADPR generation and subsequent TRPM2 channel activation by ROS and other OS-inducing pathological stimuli. PARP inhibitors have been used in many studies as an alternative or additional means to prevent OS-induced TRPM2-mediated effects [ 36 , 39 , 48 , 74 , 75 , 82 , [121] , [122] , [123] , [124] , [125] , [126] ], including cell death. Example PARP inhibitors include SB-750139, N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-N,N-dimethylacetamide (PJ34), 3-aminobenzamide (3-AB) and 3,4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H)-isoquinolinone (DPQ).…”

“…Both MPP + -induced cell death and activation of the apoptotic pathways were also inhibited by treatment with TRPM2-siRNA [ 44 ]. In a more recent study using PI staining, we have shown that exposure to H 2 O 2 (100–500 μM for 24 h) induced concentration-dependent cell death in SH-SY5Y cells and that H 2 O 2 -induced cell death was suppressed or abolished by treatment with 2-APB, PJ34 or DPQ [ 123 ]. Furthermore, by generating a stable SH-SY5Y cell line overexpressing the TRPM2 channel and comparing cell death following exposure to H 2 O 2 (30–300 μM for 24 h), we further demonstrated that an increase in TRPM2 channel expression remarkably enhanced susceptibility to H 2 O 2 -induced cell death [ 123 ].…”

“…In a more recent study using PI staining, we have shown that exposure to H 2 O 2 (100–500 μM for 24 h) induced concentration-dependent cell death in SH-SY5Y cells and that H 2 O 2 -induced cell death was suppressed or abolished by treatment with 2-APB, PJ34 or DPQ [ 123 ]. Furthermore, by generating a stable SH-SY5Y cell line overexpressing the TRPM2 channel and comparing cell death following exposure to H 2 O 2 (30–300 μM for 24 h), we further demonstrated that an increase in TRPM2 channel expression remarkably enhanced susceptibility to H 2 O 2 -induced cell death [ 123 ]. Taken together, these studies examining TRPM2 expression in the SNpc of PD brains, the SNpc of MPTP-induced PD mouse brains and also MPP + -exposed human SH-SY5Y cells, particularly the role of the TRPM2 channel in H 2 O 2 /MPP + -induced cell death in SH-SY5Y cells, suggest that PD-related OS induces TRPM2 channel expression upregulation and activation, resulting in altered intracellular Ca 2+ homeostasis and subsequent activation of calpain and the intrinsic apoptotic pathway ( Fig.…”

Roles of NAD+ and Its Metabolites Regulated Calcium Channels in Cancer

“…However, the data in cancer models mostly supports the conclusion that TRPM2 expression and function have an important role in preserving cancer cell viability and survival. Consistent with this view, the TRPM2 channel has been found to be highly expressed in numerous cancers including bladder, breast, head and neck, lung, pancreatic, prostate, melanoma, and neuroblastoma [ 51 ], among which most studies were focused on the neuroblastoma [ 58 , 59 , 60 , 61 , 62 , 63 , 64 ] ( Figure 2 ). The higher levels of ROS in cancer cells impel the enhancement of their anti-oxidant capacity to detoxify ROS and preserve cells viability.…”