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LY294002 induces differentiation and inhibits invasion of glioblastoma cells by targeting GSK-3beta and MMP.

Tian Q, Cui H, Li Y, Lu H - EXCLI J (2012)

Bottom Line: Small interfering RNA against glycogen synthase kinase-3β (GSK-3β) suppresses the induced-differentiation and invasiveness in C6 cells.LY294002 also inhibits MMP-9 expression and invasion of C6 cells, assembling the role of metalloprotease (MMP) inhibitor AG3340.Taken together, these findings suggest differentiation-inducing and invasion-inhibitory effectiveness of LY294002 in glioblastomas, most likely involving inhibition of GSK-3β and MMP respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Tianjin Children's Hospital, Tianjin 300074, China.

ABSTRACT
Glioblastomas are the most common and devastating primary tumors of the central nervous system, with high proliferative capacity, aggressive invasion, and resistance to conventional therapies. Differentiation therapy has emerged as a promising candidate modality. Here we show that the traditional phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 is capable of inducing differentiation of C6 glioblastoma cells characterized by morphological changes to astrocytic phenotype, increase in differentiation marker protein glial fibrillary acidic protein and inhibition of proliferation. Small interfering RNA against glycogen synthase kinase-3β (GSK-3β) suppresses the induced-differentiation and invasiveness in C6 cells. LY294002 also inhibits MMP-9 expression and invasion of C6 cells, assembling the role of metalloprotease (MMP) inhibitor AG3340. Taken together, these findings suggest differentiation-inducing and invasion-inhibitory effectiveness of LY294002 in glioblastomas, most likely involving inhibition of GSK-3β and MMP respectively.

No MeSH data available.


Related in: MedlinePlus

Silencing GSK-3β blocks differentiation induced by LY294002 in C6 cells. (A) Immunoblot of the GSK-3β protein levels after transfection with 10 nM scrambled (Scram) or GSK-3β siRNAs (siGSK-3β) for 36 h. (B-C) Morphology (Scale bar = 50 μM). (B) and immunoblot of the GFAP and PCNA levels (C) in GSK-3β knockdown cells subsequently stimulated with 20 μM LY294002 for 48 h.
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Figure 4: Silencing GSK-3β blocks differentiation induced by LY294002 in C6 cells. (A) Immunoblot of the GSK-3β protein levels after transfection with 10 nM scrambled (Scram) or GSK-3β siRNAs (siGSK-3β) for 36 h. (B-C) Morphology (Scale bar = 50 μM). (B) and immunoblot of the GFAP and PCNA levels (C) in GSK-3β knockdown cells subsequently stimulated with 20 μM LY294002 for 48 h.

Mentions: To further confirm the role of GSK-3β in glioblastoma cell differentiation, we examined the effects of GSK-3β inhibition. To selectively down-regulate expression of GSK-3β proteins, we evaluated the effect of isoform-specific GSK-3β gene silencing mediated by siRNAs. As shown in Figure 4A(Fig. 4), cells transfected with GSK-3β siRNA resulted in dramatically knockdown of the protein levels compared with those transfected with scrambled control siRNA. In addition, morphological transformation and elevation of GFAP during C6 cell differentiation were blocked by 10 nM GSK-3β specific siRNA (Figure 4B and 4C(Fig. 4)). These results suggest the requisite role of GSK-3β in the differentiation of glioblastoma cells and suggest that GSK-3β positively regulates the differentiation induced by LY294002.


LY294002 induces differentiation and inhibits invasion of glioblastoma cells by targeting GSK-3beta and MMP.

Tian Q, Cui H, Li Y, Lu H - EXCLI J (2012)

Silencing GSK-3β blocks differentiation induced by LY294002 in C6 cells. (A) Immunoblot of the GSK-3β protein levels after transfection with 10 nM scrambled (Scram) or GSK-3β siRNAs (siGSK-3β) for 36 h. (B-C) Morphology (Scale bar = 50 μM). (B) and immunoblot of the GFAP and PCNA levels (C) in GSK-3β knockdown cells subsequently stimulated with 20 μM LY294002 for 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920038&req=5

Figure 4: Silencing GSK-3β blocks differentiation induced by LY294002 in C6 cells. (A) Immunoblot of the GSK-3β protein levels after transfection with 10 nM scrambled (Scram) or GSK-3β siRNAs (siGSK-3β) for 36 h. (B-C) Morphology (Scale bar = 50 μM). (B) and immunoblot of the GFAP and PCNA levels (C) in GSK-3β knockdown cells subsequently stimulated with 20 μM LY294002 for 48 h.
Mentions: To further confirm the role of GSK-3β in glioblastoma cell differentiation, we examined the effects of GSK-3β inhibition. To selectively down-regulate expression of GSK-3β proteins, we evaluated the effect of isoform-specific GSK-3β gene silencing mediated by siRNAs. As shown in Figure 4A(Fig. 4), cells transfected with GSK-3β siRNA resulted in dramatically knockdown of the protein levels compared with those transfected with scrambled control siRNA. In addition, morphological transformation and elevation of GFAP during C6 cell differentiation were blocked by 10 nM GSK-3β specific siRNA (Figure 4B and 4C(Fig. 4)). These results suggest the requisite role of GSK-3β in the differentiation of glioblastoma cells and suggest that GSK-3β positively regulates the differentiation induced by LY294002.

Bottom Line: Small interfering RNA against glycogen synthase kinase-3β (GSK-3β) suppresses the induced-differentiation and invasiveness in C6 cells.LY294002 also inhibits MMP-9 expression and invasion of C6 cells, assembling the role of metalloprotease (MMP) inhibitor AG3340.Taken together, these findings suggest differentiation-inducing and invasion-inhibitory effectiveness of LY294002 in glioblastomas, most likely involving inhibition of GSK-3β and MMP respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Tianjin Children's Hospital, Tianjin 300074, China.

ABSTRACT
Glioblastomas are the most common and devastating primary tumors of the central nervous system, with high proliferative capacity, aggressive invasion, and resistance to conventional therapies. Differentiation therapy has emerged as a promising candidate modality. Here we show that the traditional phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 is capable of inducing differentiation of C6 glioblastoma cells characterized by morphological changes to astrocytic phenotype, increase in differentiation marker protein glial fibrillary acidic protein and inhibition of proliferation. Small interfering RNA against glycogen synthase kinase-3β (GSK-3β) suppresses the induced-differentiation and invasiveness in C6 cells. LY294002 also inhibits MMP-9 expression and invasion of C6 cells, assembling the role of metalloprotease (MMP) inhibitor AG3340. Taken together, these findings suggest differentiation-inducing and invasion-inhibitory effectiveness of LY294002 in glioblastomas, most likely involving inhibition of GSK-3β and MMP respectively.

No MeSH data available.


Related in: MedlinePlus