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Overexpression of candidate tumor suppressor ECRG4 inhibits glioma proliferation and invasion.

Li W, Liu X, Zhang B, Qi D, Zhang L, Jin Y, Yang H - J. Exp. Clin. Cancer Res. (2010)

Bottom Line: Cells transfected with ECRG4 showed significantly decreased cell proliferation as evaluated by MTT and colony formation assays.Furthermore, overexpression inhibited cell migration and invasion in transwell and Boyden chamber experiments and retarded the cell cycle progression from G1 to S phase by FACSCaliber cytometry.Protein levels of nuclear transcription factor NF-kB, which is involved in cell proliferation, inversely correlated with ECRG4 expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Key Laboratory of Pathobiology, Ministry of Education, Jilin University, 130021, PR China.

ABSTRACT

Background: ECRG4 has been shown to be a candidate tumor suppressor in several tumors, but its role in glioma remains poorly understood. In this study, we examined the mRNA expression of ECRG4 and investigated its biological role in glioma cells.

Methods: Real-time PCR was used to examine expression of ECRG4 in gliomas and their matched brain tissues. The effect of ECRG4 expression on cell proliferation, invasion, and migration was investigated in human U251 glioma cells. Finally, the regulation of transcription factor NF-kB by ECRG4 was evaluated by western blotting.

Results: Of the 10 paired samples analyzed, 9 glioma tissues displayed the decreased expression of ECRG4 compared to matched normal brain tissues. Cells transfected with ECRG4 showed significantly decreased cell proliferation as evaluated by MTT and colony formation assays. Furthermore, overexpression inhibited cell migration and invasion in transwell and Boyden chamber experiments and retarded the cell cycle progression from G1 to S phase by FACSCaliber cytometry. Protein levels of nuclear transcription factor NF-kB, which is involved in cell proliferation, inversely correlated with ECRG4 expression.

Conclusion: Our data suggest that ECRG4 serves as a tumor suppressor in glioma.

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Related in: MedlinePlus

Restored expression of ECRG4 in glioma U251 cells. A. Real-time PCR analysis indicated the highest mRNA expression of ECRG4 in two cell clones pEGFP-ECRG4-5 and -7. B. Western blotting assay shows significantly increased protein expression of ECRG4 in pEGFP-ECRG4-5 and -7 comparing to Control cells. β-actin was used as the internal control.
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Figure 2: Restored expression of ECRG4 in glioma U251 cells. A. Real-time PCR analysis indicated the highest mRNA expression of ECRG4 in two cell clones pEGFP-ECRG4-5 and -7. B. Western blotting assay shows significantly increased protein expression of ECRG4 in pEGFP-ECRG4-5 and -7 comparing to Control cells. β-actin was used as the internal control.

Mentions: To study the biological functions of ECRG4, we introduced ECRG4 into U251 glioma cells using a pEGFP-N1 eucaryotic expression vector containing ECRG4 gene. Seven stably transfected cell clones were obtained. Real-time PCR identified two cell clones(ECRG4-5,-7) with the highest mRNA expression of ECRG4(Figure 2A). Further, Western blotting assay with a GFP antibody showed that ECRG4-GFP fusion protein in two cell clones was highly expressed, compared to control clone cells (Figure 2B).


Overexpression of candidate tumor suppressor ECRG4 inhibits glioma proliferation and invasion.

Li W, Liu X, Zhang B, Qi D, Zhang L, Jin Y, Yang H - J. Exp. Clin. Cancer Res. (2010)

Restored expression of ECRG4 in glioma U251 cells. A. Real-time PCR analysis indicated the highest mRNA expression of ECRG4 in two cell clones pEGFP-ECRG4-5 and -7. B. Western blotting assay shows significantly increased protein expression of ECRG4 in pEGFP-ECRG4-5 and -7 comparing to Control cells. β-actin was used as the internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2913949&req=5

Figure 2: Restored expression of ECRG4 in glioma U251 cells. A. Real-time PCR analysis indicated the highest mRNA expression of ECRG4 in two cell clones pEGFP-ECRG4-5 and -7. B. Western blotting assay shows significantly increased protein expression of ECRG4 in pEGFP-ECRG4-5 and -7 comparing to Control cells. β-actin was used as the internal control.
Mentions: To study the biological functions of ECRG4, we introduced ECRG4 into U251 glioma cells using a pEGFP-N1 eucaryotic expression vector containing ECRG4 gene. Seven stably transfected cell clones were obtained. Real-time PCR identified two cell clones(ECRG4-5,-7) with the highest mRNA expression of ECRG4(Figure 2A). Further, Western blotting assay with a GFP antibody showed that ECRG4-GFP fusion protein in two cell clones was highly expressed, compared to control clone cells (Figure 2B).

Bottom Line: Cells transfected with ECRG4 showed significantly decreased cell proliferation as evaluated by MTT and colony formation assays.Furthermore, overexpression inhibited cell migration and invasion in transwell and Boyden chamber experiments and retarded the cell cycle progression from G1 to S phase by FACSCaliber cytometry.Protein levels of nuclear transcription factor NF-kB, which is involved in cell proliferation, inversely correlated with ECRG4 expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Key Laboratory of Pathobiology, Ministry of Education, Jilin University, 130021, PR China.

ABSTRACT

Background: ECRG4 has been shown to be a candidate tumor suppressor in several tumors, but its role in glioma remains poorly understood. In this study, we examined the mRNA expression of ECRG4 and investigated its biological role in glioma cells.

Methods: Real-time PCR was used to examine expression of ECRG4 in gliomas and their matched brain tissues. The effect of ECRG4 expression on cell proliferation, invasion, and migration was investigated in human U251 glioma cells. Finally, the regulation of transcription factor NF-kB by ECRG4 was evaluated by western blotting.

Results: Of the 10 paired samples analyzed, 9 glioma tissues displayed the decreased expression of ECRG4 compared to matched normal brain tissues. Cells transfected with ECRG4 showed significantly decreased cell proliferation as evaluated by MTT and colony formation assays. Furthermore, overexpression inhibited cell migration and invasion in transwell and Boyden chamber experiments and retarded the cell cycle progression from G1 to S phase by FACSCaliber cytometry. Protein levels of nuclear transcription factor NF-kB, which is involved in cell proliferation, inversely correlated with ECRG4 expression.

Conclusion: Our data suggest that ECRG4 serves as a tumor suppressor in glioma.

Show MeSH
Related in: MedlinePlus