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Essential role of GATA3 in regulation of differentiation and cell proliferation in SK-N-SH neuroblastoma cells.

Peng H, Ke XX, Hu R, Yang L, Cui H, Wei Y - Mol Med Rep (2014)

Bottom Line: Neuroblastoma is a common solid malignant tumor of the sympathetic nervous system, which contributes to 15% of cancer‑related mortality in children.The results demonstrated that GATA3 is a prognostic marker for survival in patients with neuroblastoma, and that high‑level GATA3 expression is associated with increased self‑renewal and proliferation of neuroblastoma cells.Retinoic acid treatment led to GATA3 downregulation together with neuronal differentiation, suppression of cell proliferation and inhibition of tumorigenecity in neuroblastoma cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cancer Biotherapy, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.

ABSTRACT
Neuroblastoma is a common solid malignant tumor of the sympathetic nervous system, which contributes to 15% of cancer‑related mortality in children. The differentiation status of neuroblastoma is correlated with clinical outcome, and the induction of differentiation thus constitutes a therapeutic approach in this disease. However, the molecular mechanisms that control the differentiation of neuroblastoma remain poorly understood. The present study aimed to define whether GATA3 is involved in the differentiation of neuroblastoma cells. The results demonstrated that GATA3 is a prognostic marker for survival in patients with neuroblastoma, and that high‑level GATA3 expression is associated with increased self‑renewal and proliferation of neuroblastoma cells. Retinoic acid treatment led to GATA3 downregulation together with neuronal differentiation, suppression of cell proliferation and inhibition of tumorigenecity in neuroblastoma cells. These findings suggest that GATA3 is a key regulator of neuroblastoma differentiation, and provide a novel potential therapeutic strategy for the induction of neuroblastoma differentiation.

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Effect of GATA3 overexpression on the proliferation and self-renewal of neuroblastoma cells. (A) Western blot analysis of GATA3 expression in SK-N-SH cells with GFP or GATA3 overexpression. α-Tubulin levels were used as a loading control. (B) SK-N-SH cells with GFP or GATA3 overexpression were analyzed for cell growth curve with an MTT assay. (C) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in six-well culture plates. At days 14 to 21, soft agar colonies grew from cells with GFP or GATA3 overexpression. Cells with GATA3 overexpression were observed to give rise to larger and more colonies in soft agar. (D) Colonies >0.5 mm or that contained >50 cells were recorded. (E) and (F) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in Matrigel ultra-low attachment plates. At days 14 to 21, spheres grew and were recorded (magnification, ×10). Data in (B), (D) and (F) are presented as the average obtained from three independent experiments. Error bars represent the standard deviation. *P≤0.05. GFP, green fluorescent protein; OD, optical density.
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f4-mmr-11-02-0881: Effect of GATA3 overexpression on the proliferation and self-renewal of neuroblastoma cells. (A) Western blot analysis of GATA3 expression in SK-N-SH cells with GFP or GATA3 overexpression. α-Tubulin levels were used as a loading control. (B) SK-N-SH cells with GFP or GATA3 overexpression were analyzed for cell growth curve with an MTT assay. (C) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in six-well culture plates. At days 14 to 21, soft agar colonies grew from cells with GFP or GATA3 overexpression. Cells with GATA3 overexpression were observed to give rise to larger and more colonies in soft agar. (D) Colonies >0.5 mm or that contained >50 cells were recorded. (E) and (F) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in Matrigel ultra-low attachment plates. At days 14 to 21, spheres grew and were recorded (magnification, ×10). Data in (B), (D) and (F) are presented as the average obtained from three independent experiments. Error bars represent the standard deviation. *P≤0.05. GFP, green fluorescent protein; OD, optical density.

Mentions: To confirm the correlation between GATA3 and self-renewal of neuroblastoma cells, GATA3 was overexpressed in neuroblastoma cells, using GFP as a control (Fig. 4A). GATA3 significantly increased cell proliferation, which was verified by MTT analysis (Fig. 4B). In addition, GATA3 markedly upregulated the self-renewal ability, including the colony forming and sphere forming capability, of neuroblastoma cells (Fig. 4C and 4F). These results demonstrated that high expression of GATA3 is associated with increased self-renewal and cell proliferation in neuroblastoma cells.


Essential role of GATA3 in regulation of differentiation and cell proliferation in SK-N-SH neuroblastoma cells.

Peng H, Ke XX, Hu R, Yang L, Cui H, Wei Y - Mol Med Rep (2014)

Effect of GATA3 overexpression on the proliferation and self-renewal of neuroblastoma cells. (A) Western blot analysis of GATA3 expression in SK-N-SH cells with GFP or GATA3 overexpression. α-Tubulin levels were used as a loading control. (B) SK-N-SH cells with GFP or GATA3 overexpression were analyzed for cell growth curve with an MTT assay. (C) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in six-well culture plates. At days 14 to 21, soft agar colonies grew from cells with GFP or GATA3 overexpression. Cells with GATA3 overexpression were observed to give rise to larger and more colonies in soft agar. (D) Colonies >0.5 mm or that contained >50 cells were recorded. (E) and (F) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in Matrigel ultra-low attachment plates. At days 14 to 21, spheres grew and were recorded (magnification, ×10). Data in (B), (D) and (F) are presented as the average obtained from three independent experiments. Error bars represent the standard deviation. *P≤0.05. GFP, green fluorescent protein; OD, optical density.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262502&req=5

f4-mmr-11-02-0881: Effect of GATA3 overexpression on the proliferation and self-renewal of neuroblastoma cells. (A) Western blot analysis of GATA3 expression in SK-N-SH cells with GFP or GATA3 overexpression. α-Tubulin levels were used as a loading control. (B) SK-N-SH cells with GFP or GATA3 overexpression were analyzed for cell growth curve with an MTT assay. (C) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in six-well culture plates. At days 14 to 21, soft agar colonies grew from cells with GFP or GATA3 overexpression. Cells with GATA3 overexpression were observed to give rise to larger and more colonies in soft agar. (D) Colonies >0.5 mm or that contained >50 cells were recorded. (E) and (F) SK-N-SH cells with GFP or GATA3 overexpression were plated at 4×103 cells per well in Matrigel ultra-low attachment plates. At days 14 to 21, spheres grew and were recorded (magnification, ×10). Data in (B), (D) and (F) are presented as the average obtained from three independent experiments. Error bars represent the standard deviation. *P≤0.05. GFP, green fluorescent protein; OD, optical density.
Mentions: To confirm the correlation between GATA3 and self-renewal of neuroblastoma cells, GATA3 was overexpressed in neuroblastoma cells, using GFP as a control (Fig. 4A). GATA3 significantly increased cell proliferation, which was verified by MTT analysis (Fig. 4B). In addition, GATA3 markedly upregulated the self-renewal ability, including the colony forming and sphere forming capability, of neuroblastoma cells (Fig. 4C and 4F). These results demonstrated that high expression of GATA3 is associated with increased self-renewal and cell proliferation in neuroblastoma cells.

Bottom Line: Neuroblastoma is a common solid malignant tumor of the sympathetic nervous system, which contributes to 15% of cancer‑related mortality in children.The results demonstrated that GATA3 is a prognostic marker for survival in patients with neuroblastoma, and that high‑level GATA3 expression is associated with increased self‑renewal and proliferation of neuroblastoma cells.Retinoic acid treatment led to GATA3 downregulation together with neuronal differentiation, suppression of cell proliferation and inhibition of tumorigenecity in neuroblastoma cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cancer Biotherapy, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.

ABSTRACT
Neuroblastoma is a common solid malignant tumor of the sympathetic nervous system, which contributes to 15% of cancer‑related mortality in children. The differentiation status of neuroblastoma is correlated with clinical outcome, and the induction of differentiation thus constitutes a therapeutic approach in this disease. However, the molecular mechanisms that control the differentiation of neuroblastoma remain poorly understood. The present study aimed to define whether GATA3 is involved in the differentiation of neuroblastoma cells. The results demonstrated that GATA3 is a prognostic marker for survival in patients with neuroblastoma, and that high‑level GATA3 expression is associated with increased self‑renewal and proliferation of neuroblastoma cells. Retinoic acid treatment led to GATA3 downregulation together with neuronal differentiation, suppression of cell proliferation and inhibition of tumorigenecity in neuroblastoma cells. These findings suggest that GATA3 is a key regulator of neuroblastoma differentiation, and provide a novel potential therapeutic strategy for the induction of neuroblastoma differentiation.

Show MeSH
Related in: MedlinePlus