Limits...
SOX4 inhibits GBM cell growth and induces G0/G1 cell cycle arrest through Akt-p53 axis.

Zhang J, Jiang H, Shao J, Mao R, Liu J, Ma Y, Fang X, Zhao N, Zheng S, Lin B - BMC Neurol (2014)

Bottom Line: High SOX4 expression was significantly associated with good prognosis of primary GBMs. SOX4 inhibited the growth of GBM cell line LN229, A172G and U87MG, partly via the activation of p53-p21 signaling and down-regulation of phosphorylated AKT1.Gene expression profiling and subsequent gene ontology analysis showed that SOX4 influenced several key pathways including the Wnt/ beta-catenin and TGF-beta signaling pathways.Our study found that SOX4 acts as a tumor suppressor in GBM cells by induce cell cycle arrest and inhibiting cell growth.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, P R China. jingzh1985@gmail.com.

ABSTRACT

Background: SOX4 is a transcription factor required for tissue development and differentiation in vertebrates. Overexpression of SOX4 has been reported in many cancers including glioblastoma multiforme (GBM), however, the underlying mechanism of actions has not been studied. In this study, we investigated the role of SOX4 in GBM.

Methods: Kaplan-Meier analysis was performed to assess the association between SOX4 expression levels and survival times in primary GBM samples. Cre/lox P system was used to generate gain or loss of SOX4 in GBM cells, and microarray analysis uncovered the regulation network of SOX4 in GBM cells.

Results: High SOX4 expression was significantly associated with good prognosis of primary GBMs. SOX4 inhibited the growth of GBM cell line LN229, A172G and U87MG, partly via the activation of p53-p21 signaling and down-regulation of phosphorylated AKT1. Gene expression profiling and subsequent gene ontology analysis showed that SOX4 influenced several key pathways including the Wnt/ beta-catenin and TGF-beta signaling pathways.

Conclusions: Our study found that SOX4 acts as a tumor suppressor in GBM cells by induce cell cycle arrest and inhibiting cell growth.

No MeSH data available.


Related in: MedlinePlus

Transient SOX4 overexpression inhibits growth of LN229 cells and induces G0/G1 cell cycle arrest. (A) LN229 cells were transiently transfected with Flag-Sox4-HA or pEGFP-N1 plasmid as control. After 48 hours, protein expression was assayed by Western blotting using anti-HA antibody (B) Cell vitality was determined by CCK-8 assay after transfection of LN229 cells with Flag-Sox4-HA plasmid or pEGFP-N1 and Cre-GFP as control at the indicated time points. Values at the indicated time points were provided as the mean cell number with an SD of eight wells. *, P < 0.05 (C) Colonies formed by Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells were shown 2 weeks after plating in 6 well plate. Upper panel showed the quantification of the colony in Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells. Values are the means ± SD of triplicate experiments, *P < 0.05 (D) Impact of SOX4 on cell cycle of LN229 cells. The percentage of cells in G0/G1, S, and G2/M phases is shown in the left panel. And the statistic analysis is also shown in the right panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4233052&req=5

Fig2: Transient SOX4 overexpression inhibits growth of LN229 cells and induces G0/G1 cell cycle arrest. (A) LN229 cells were transiently transfected with Flag-Sox4-HA or pEGFP-N1 plasmid as control. After 48 hours, protein expression was assayed by Western blotting using anti-HA antibody (B) Cell vitality was determined by CCK-8 assay after transfection of LN229 cells with Flag-Sox4-HA plasmid or pEGFP-N1 and Cre-GFP as control at the indicated time points. Values at the indicated time points were provided as the mean cell number with an SD of eight wells. *, P < 0.05 (C) Colonies formed by Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells were shown 2 weeks after plating in 6 well plate. Upper panel showed the quantification of the colony in Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells. Values are the means ± SD of triplicate experiments, *P < 0.05 (D) Impact of SOX4 on cell cycle of LN229 cells. The percentage of cells in G0/G1, S, and G2/M phases is shown in the left panel. And the statistic analysis is also shown in the right panel.

Mentions: We started with testing the functional consequences of SOX4 overexpression using a transient expression system. We transiently transfected GBM cell line LN229 with the Flag-SOX4-HA construct and we used the empty vector pEGFP-N1 construct without the HA tag as the control. Forty-eight hours after transfection, we detected marked increase of SOX4 protein expression in LN229 cells by western blot with anti-HA antibody (Figure 2A), as shown by the band at 70 KDa for the Flag-SOX4-HA protein [10,28].Figure 2


SOX4 inhibits GBM cell growth and induces G0/G1 cell cycle arrest through Akt-p53 axis.

Zhang J, Jiang H, Shao J, Mao R, Liu J, Ma Y, Fang X, Zhao N, Zheng S, Lin B - BMC Neurol (2014)

Transient SOX4 overexpression inhibits growth of LN229 cells and induces G0/G1 cell cycle arrest. (A) LN229 cells were transiently transfected with Flag-Sox4-HA or pEGFP-N1 plasmid as control. After 48 hours, protein expression was assayed by Western blotting using anti-HA antibody (B) Cell vitality was determined by CCK-8 assay after transfection of LN229 cells with Flag-Sox4-HA plasmid or pEGFP-N1 and Cre-GFP as control at the indicated time points. Values at the indicated time points were provided as the mean cell number with an SD of eight wells. *, P < 0.05 (C) Colonies formed by Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells were shown 2 weeks after plating in 6 well plate. Upper panel showed the quantification of the colony in Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells. Values are the means ± SD of triplicate experiments, *P < 0.05 (D) Impact of SOX4 on cell cycle of LN229 cells. The percentage of cells in G0/G1, S, and G2/M phases is shown in the left panel. And the statistic analysis is also shown in the right panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4233052&req=5

Fig2: Transient SOX4 overexpression inhibits growth of LN229 cells and induces G0/G1 cell cycle arrest. (A) LN229 cells were transiently transfected with Flag-Sox4-HA or pEGFP-N1 plasmid as control. After 48 hours, protein expression was assayed by Western blotting using anti-HA antibody (B) Cell vitality was determined by CCK-8 assay after transfection of LN229 cells with Flag-Sox4-HA plasmid or pEGFP-N1 and Cre-GFP as control at the indicated time points. Values at the indicated time points were provided as the mean cell number with an SD of eight wells. *, P < 0.05 (C) Colonies formed by Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells were shown 2 weeks after plating in 6 well plate. Upper panel showed the quantification of the colony in Flag-Sox4-HA or pEGFP-N1or Cre-GFP infected LN229 cells. Values are the means ± SD of triplicate experiments, *P < 0.05 (D) Impact of SOX4 on cell cycle of LN229 cells. The percentage of cells in G0/G1, S, and G2/M phases is shown in the left panel. And the statistic analysis is also shown in the right panel.
Mentions: We started with testing the functional consequences of SOX4 overexpression using a transient expression system. We transiently transfected GBM cell line LN229 with the Flag-SOX4-HA construct and we used the empty vector pEGFP-N1 construct without the HA tag as the control. Forty-eight hours after transfection, we detected marked increase of SOX4 protein expression in LN229 cells by western blot with anti-HA antibody (Figure 2A), as shown by the band at 70 KDa for the Flag-SOX4-HA protein [10,28].Figure 2

Bottom Line: High SOX4 expression was significantly associated with good prognosis of primary GBMs. SOX4 inhibited the growth of GBM cell line LN229, A172G and U87MG, partly via the activation of p53-p21 signaling and down-regulation of phosphorylated AKT1.Gene expression profiling and subsequent gene ontology analysis showed that SOX4 influenced several key pathways including the Wnt/ beta-catenin and TGF-beta signaling pathways.Our study found that SOX4 acts as a tumor suppressor in GBM cells by induce cell cycle arrest and inhibiting cell growth.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, P R China. jingzh1985@gmail.com.

ABSTRACT

Background: SOX4 is a transcription factor required for tissue development and differentiation in vertebrates. Overexpression of SOX4 has been reported in many cancers including glioblastoma multiforme (GBM), however, the underlying mechanism of actions has not been studied. In this study, we investigated the role of SOX4 in GBM.

Methods: Kaplan-Meier analysis was performed to assess the association between SOX4 expression levels and survival times in primary GBM samples. Cre/lox P system was used to generate gain or loss of SOX4 in GBM cells, and microarray analysis uncovered the regulation network of SOX4 in GBM cells.

Results: High SOX4 expression was significantly associated with good prognosis of primary GBMs. SOX4 inhibited the growth of GBM cell line LN229, A172G and U87MG, partly via the activation of p53-p21 signaling and down-regulation of phosphorylated AKT1. Gene expression profiling and subsequent gene ontology analysis showed that SOX4 influenced several key pathways including the Wnt/ beta-catenin and TGF-beta signaling pathways.

Conclusions: Our study found that SOX4 acts as a tumor suppressor in GBM cells by induce cell cycle arrest and inhibiting cell growth.

No MeSH data available.


Related in: MedlinePlus