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Transglutaminase-2 Is Involved in All-Trans Retinoic Acid-Induced Invasion and Matrix Metalloproteinases Expression of SH-SY5Y Neuroblastoma Cells via NF-κB Pathway.

Lee HJ, Park MK, Bae HC, Yoon HJ, Kim SY, Lee CH - Biomol Ther (Seoul) (2012)

Bottom Line: ATRA induced the activation of NF-κB in the SH-SY5Y cells and CTM suppressed the activation of NF-κB.Gene silencing of Tgase-2 suppressed the MMP expression by ATRA.These results suggested that Tgase-2 might be a new target for controlling the ATRA-induced invasion of NBs.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dongguk University, Seoul 100-715.

ABSTRACT
All-trans retinoic acid (ATRA) is currently used in adjuvant differentiation-based treatment of residual or relapsed neuroblastoma (NB). It has been reported that short-term ATRA treatment induces migration and invasion of SH-SY5Y via transglutaminase-2 (Tgase-2). However, the detailed mechanism of Tgase-2's involvement in NB cell invasion remains unclear. Therefore we investigated the role of Tgase-2 in invasion of NB cells using SH-SY5Y cells. ATRA dose-dependently induced the invasion of SH-SY5Y cells. Cystamine (CTM), a well known tgase inhibitor suppressed the ATRA-induced invasion of SH-SY5Y cells in a dose-dependent manner. Matrix metalloproteinase-9 (MMP-9) and MMP-2, well known genes involved in invasion of cancer cells were induced in the ATRA-induced invasion of the SH-SH5Y cells. Treatment of CTM suppressed the MMP-9 and MMP-2 enzyme activities in the ATRA-induced invasion of the SH-SY5Y cells. To confirm the involvement of Tgase-2, gene silencing of Tgase-2 was performed in the ATRA-induced invasion of the SH-SH5Y cells. The siRNA of Tgase-2 suppressed the MMP-9 and MMP-2 activity of the SH-SY5Y cells. MMP-2 and MMP-9 are well known target genes of NF-κB. Therefore the relationship of Tgase-2 and NF-κB in the ATRA-induced invasion of the SH-SY5Y cells was examined using siRNA and CTM. ATRA induced the activation of NF-κB in the SH-SY5Y cells and CTM suppressed the activation of NF-κB. Gene silencing of Tgase-2 suppressed the MMP expression by ATRA. These results suggested that Tgase-2 might be a new target for controlling the ATRA-induced invasion of NBs.

No MeSH data available.


Related in: MedlinePlus

Activation of ATRA-induced NF-κB and suppression of ATRA-induced NF-κB activation by CTM or gene silencing. (A) Western blot of Tgase-2 and IκBα in ATRA-induced invasion of SH-SY5Y cells. (B) Western blot of NF-κB p65 in nucleus fraction and IκBα in cytosol fraction of ATRA-treated SH-SY5Y cells (w/wo CTM at 100 μM). (C) Western blot of NF-κB p65 and Tgase-2 in nucleus fraction and IκBα and Tgase-2 in cytosol fraction in SH-SY5Y and SH-SY5YShTG2 with or without ATRA. (D) Proposed scheme of involvement of Tgase-2 in ATRA-induced invasion of SH-SY5Y cells.
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Figure 004: Activation of ATRA-induced NF-κB and suppression of ATRA-induced NF-κB activation by CTM or gene silencing. (A) Western blot of Tgase-2 and IκBα in ATRA-induced invasion of SH-SY5Y cells. (B) Western blot of NF-κB p65 in nucleus fraction and IκBα in cytosol fraction of ATRA-treated SH-SY5Y cells (w/wo CTM at 100 μM). (C) Western blot of NF-κB p65 and Tgase-2 in nucleus fraction and IκBα and Tgase-2 in cytosol fraction in SH-SY5Y and SH-SY5YShTG2 with or without ATRA. (D) Proposed scheme of involvement of Tgase-2 in ATRA-induced invasion of SH-SY5Y cells.

Mentions: expression of Tgase-2 (Fig. 4A).


Transglutaminase-2 Is Involved in All-Trans Retinoic Acid-Induced Invasion and Matrix Metalloproteinases Expression of SH-SY5Y Neuroblastoma Cells via NF-κB Pathway.

Lee HJ, Park MK, Bae HC, Yoon HJ, Kim SY, Lee CH - Biomol Ther (Seoul) (2012)

Activation of ATRA-induced NF-κB and suppression of ATRA-induced NF-κB activation by CTM or gene silencing. (A) Western blot of Tgase-2 and IκBα in ATRA-induced invasion of SH-SY5Y cells. (B) Western blot of NF-κB p65 in nucleus fraction and IκBα in cytosol fraction of ATRA-treated SH-SY5Y cells (w/wo CTM at 100 μM). (C) Western blot of NF-κB p65 and Tgase-2 in nucleus fraction and IκBα and Tgase-2 in cytosol fraction in SH-SY5Y and SH-SY5YShTG2 with or without ATRA. (D) Proposed scheme of involvement of Tgase-2 in ATRA-induced invasion of SH-SY5Y cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3794525&req=5

Figure 004: Activation of ATRA-induced NF-κB and suppression of ATRA-induced NF-κB activation by CTM or gene silencing. (A) Western blot of Tgase-2 and IκBα in ATRA-induced invasion of SH-SY5Y cells. (B) Western blot of NF-κB p65 in nucleus fraction and IκBα in cytosol fraction of ATRA-treated SH-SY5Y cells (w/wo CTM at 100 μM). (C) Western blot of NF-κB p65 and Tgase-2 in nucleus fraction and IκBα and Tgase-2 in cytosol fraction in SH-SY5Y and SH-SY5YShTG2 with or without ATRA. (D) Proposed scheme of involvement of Tgase-2 in ATRA-induced invasion of SH-SY5Y cells.
Mentions: expression of Tgase-2 (Fig. 4A).

Bottom Line: ATRA induced the activation of NF-κB in the SH-SY5Y cells and CTM suppressed the activation of NF-κB.Gene silencing of Tgase-2 suppressed the MMP expression by ATRA.These results suggested that Tgase-2 might be a new target for controlling the ATRA-induced invasion of NBs.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dongguk University, Seoul 100-715.

ABSTRACT
All-trans retinoic acid (ATRA) is currently used in adjuvant differentiation-based treatment of residual or relapsed neuroblastoma (NB). It has been reported that short-term ATRA treatment induces migration and invasion of SH-SY5Y via transglutaminase-2 (Tgase-2). However, the detailed mechanism of Tgase-2's involvement in NB cell invasion remains unclear. Therefore we investigated the role of Tgase-2 in invasion of NB cells using SH-SY5Y cells. ATRA dose-dependently induced the invasion of SH-SY5Y cells. Cystamine (CTM), a well known tgase inhibitor suppressed the ATRA-induced invasion of SH-SY5Y cells in a dose-dependent manner. Matrix metalloproteinase-9 (MMP-9) and MMP-2, well known genes involved in invasion of cancer cells were induced in the ATRA-induced invasion of the SH-SH5Y cells. Treatment of CTM suppressed the MMP-9 and MMP-2 enzyme activities in the ATRA-induced invasion of the SH-SY5Y cells. To confirm the involvement of Tgase-2, gene silencing of Tgase-2 was performed in the ATRA-induced invasion of the SH-SH5Y cells. The siRNA of Tgase-2 suppressed the MMP-9 and MMP-2 activity of the SH-SY5Y cells. MMP-2 and MMP-9 are well known target genes of NF-κB. Therefore the relationship of Tgase-2 and NF-κB in the ATRA-induced invasion of the SH-SY5Y cells was examined using siRNA and CTM. ATRA induced the activation of NF-κB in the SH-SY5Y cells and CTM suppressed the activation of NF-κB. Gene silencing of Tgase-2 suppressed the MMP expression by ATRA. These results suggested that Tgase-2 might be a new target for controlling the ATRA-induced invasion of NBs.

No MeSH data available.


Related in: MedlinePlus