In this study we show that anti-tumor effect of sulforaphane (SFN) is partially realized through the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). This effect was verified in vitro on three different stable cell lines and also in vivo on the model of nude mice with developed tumors. Early response (6 hours) of A2780 ovarian carcinoma cells to SFN treatment involves generation of mitochondrial ROS and increased transcription of NRF2 and its downstream regulated genes including heme oxygenase 1, NAD(P)H:quinine oxidoreductase 1, and KLF9. Prolonged SFN treatment (24 hours) upregulated expression of NRF2 and IP3R1. SFN induces a time-dependent phosphorylation wave of HSP27. Use of IP3R inhibitor Xestospongin C (Xest) attenuates both SFN-induced apoptosis and the level of NRF2 protein expression. In addition, Xest partially attenuates anti-tumor effect of SFN in vivo. SFN-induced apoptosis is completely inhibited by silencing of IP3R1 gene but only partially blocked by silencing of NRF2; silencing of IP3R2 and IP3R3 had no effect on these cells. Xest inhibitor does not significantly modify SFN-induced increase in the rapid activity of ARE and AP1 responsive elements. We found that Xest effectively reverses the SFN-dependent increase of nuclear content and decrease of reticular calcium content. In addition, immunofluorescent staining with IP3R1 antibody revealed that SFN treatment induces translocation of IP3R1 to the nucleus. Our results clearly show that IP3R1 is involved in SFN-induced apoptosis through the depletion of reticular calcium and modulation of transcription factors through nuclear calcium up-regulation.

• MeSH
• aktivace transkripce účinky léků   MeSH
• antikarcinogenní látky farmakologie   terapeutické užití   MeSH
• antioxidační responzivní elementy MeSH
• apoptóza účinky léků   MeSH
• buněčné jádro metabolismus   MeSH
• endoplazmatické retikulum metabolismus   MeSH
• faktor 2 související s NF-E2 metabolismus   MeSH
• hemoxygenasa-1 metabolismus   MeSH
• inositol-1,4,5-trisfosfát - receptory antagonisté a inhibitory   metabolismus   MeSH
• isothiokyanatany farmakologie   terapeutické užití   MeSH
• lidé MeSH
• makrocyklické sloučeniny farmakologie   MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• myši nahé MeSH
• myši MeSH
• NAD(P)H dehydrogenasa (chinon) metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory vaječníků farmakoterapie   patologie   MeSH
• oxazoly farmakologie   MeSH
• oxidační stres účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• transkripční faktory Krüppel-like metabolismus   MeSH
• upregulace MeSH
• vápník metabolismus   MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Cellular differentiation is the process, by which a cell changes from one cell type to another, preferentially to the more specialized one. Calcium fluxes play an important role in this action. Differentiated NG108-15 or PC12 cells serve as models for studying neuronal pathways. NG108-15 cell line is a reliable model of cholinergic neuronal cells. These cells differentiate to a neuronal phenotype due to the dibutyryl cAMP (dbcAMP) treatment. We have shown that a slow sulfide donor - GYY4137 - can also act as a differentiating factor in NG108-15 cell line. Calcium is an unavoidable ion required in NG108-15 cell differentiation by both, dbcAMP and GYY4137, since cultivation in EGTA completely prevented differentiation of these cells. In this work we focused primarily on the role of reticular calcium in the process of NG108-15 cell differentiation. We have found that dbcAMP and also GYY4137 decreased reticular calcium concentration by different mechanisms. GYY4137 caused a rapid decrease in type 2 sarco/endoplasmic calcium ATPase (SERCA2) mRNA and protein, which results in lower calcium levels in the endoplasmic reticulum compared to the control, untreated group. The dbcAMP revealed rapid increase in expression of the type 3 IP3 receptor, which participates in a calcium clearance from the endoplasmic reticulum. These results point to the important role of reticular calcium in a NG108-15 cell differentiation.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• dibutyryl cyklický AMP aplikace a dávkování   MeSH
• inositol-1,4,5-trisfosfát - receptory metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• morfoliny aplikace a dávkování   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• neurony účinky léků   fyziologie   MeSH
• organothiofosforové sloučeniny aplikace a dávkování   MeSH
• sarkoplazmatická Ca2+-ATPáza metabolismus   MeSH
• sulfan aplikace a dávkování   MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH