Silencing of Insulin-like Growth Factor-binding Protein-2 in Human Glioblastoma Cells Reduces Both Invasiveness and Expression of Progression-associated Gene CD24

Abstract

Glioblastoma multiforme (GBM) is a malignant brain tumor characterized by rapid growth and extensive invasiveness. Overexpression of insulin-like growth factor-binding protein-2 (IGFBP-2) has been reported in GBM. However, it remains to be determined how IGFBP-2 is involved in the progression of GBM. We utilized short hairpin-RNA (shRNA) expression retroviral vectors to inactivate the IGFBP-2 gene permanently in two human GBM cell and of IGFBP-2 in of the in and in of by of gene has been in progression of in GBM and the of in GBM the of to of GBM cell and a human GBM cell the IGFBP-2 and of IGFBP-2 the and in in by IGFBP-2 expression is by IGFBP-2 and to of GBM Glioblastoma multiforme (GBM) is a malignant brain tumor characterized by rapid growth and extensive invasiveness. Overexpression of insulin-like growth factor-binding protein-2 (IGFBP-2) has been reported in GBM. However, it remains to be determined how IGFBP-2 is involved in the progression of GBM. We utilized short hairpin-RNA (shRNA) expression retroviral vectors to inactivate the IGFBP-2 gene permanently in two human GBM cell and of IGFBP-2 in of the in and in of by of gene has been in progression of in GBM and the of in GBM the of to of GBM cell and a human GBM cell the IGFBP-2 and of IGFBP-2 the and in in by IGFBP-2 expression is by IGFBP-2 and to of GBM the of brain Glioblastoma multiforme (GBM) insulin-like growth factor-binding insulin-like growth short insulin-like growth factor-binding insulin-like growth short is the malignant and is characterized by rapid extensive and extensive of the tumor be by is the of the of GBM remains and the is to the of GBM and to of and of GBM of the of GBM and of growth factor-binding a of the of insulin-like growth in the been been to the a of be to tumor progression IGFBP-2 is a is in the expression expression of IGFBP-2 has been reported in and IGFBP-2 in malignant in expression of IGFBP-2 tumor and IGFBP-2 by GBM by However, it remains to be determined how IGFBP-2 is involved in the growth and of tumor and IGFBP-2 the of tumor of GBM to the expression of is a is to the cell has is a a has been the of in tumor in to be involved in cell and is in a of and and a of the expression of is a is the expression of in of in in of the in However, is of the the expression of and in tumor the of IGFBP-2 in GBM in and in and the by IGFBP-2 in human GBM of two human GBM cell in the IGFBP-2 gene permanently by retroviral expression of short involved in of GBM and of of GBM the and the by the of the of of human GBM cell and and in in a in and the cell the and the of and and a the of in cell of growth and the of and by and of human brain of a of and of and of the of and and of and and and and and and and and by IGFBP-2 and in a the to the IGFBP-2 the the of the and of of the IGFBP-2 and IGFBP-2 and and and of the and in the of and of and GBM by a expression retroviral (IGFBP-2) IGFBP-2 to and to of a retroviral expression of IGFBP-2 human IGFBP-2 a retroviral expression of retroviral in of retroviral and of the in and the the retroviral a and to GBM in the in the of and in a the a of and and a in cell of IGFBP-2 in IGFBP-2 GBM of IGFBP-2 expression in IGFBP-2 the expression the expression and to the and in and human by in the in cell and of by and in the by in and a in a IGFBP-2 and and and of in of GBM a to a a and a by the a the the a of the the of in by of a the of in the and the the of the a the the by the in of GBM by the of in GBM of the of and of the the brain brain in in and the of by in of to human IGFBP-2 to the of GBM and the by a by and by the the to and and the expression the of the gene the a IGFBP-2 in a in the and the and by a gene the and the the two of of IGFBP-2 and in the expression of and IGFBP-2 in human GBM and GBM cell and brain and in GBM and of the cell and in cell expression of IGFBP-2 in brain the in GBM in brain IGFBP-2 in of human GBM cell expression in of GBM of IGFBP-2 the of IGFBP-2 in GBM cell IGFBP-2 retroviral vectors of a the IGFBP-2 of and IGFBP-2 of two human GBM cell and the of retroviral of expression by a IGFBP-2 in a in the IGFBP-2 in IGFBP-2 the of IGFBP-2 in the expression of IGFBP-2 a We of the of IGFBP-2 in of by the of it in and the of by the of IGFBP-2 is the IGFBP-2 the of to the the IGFBP-2 gene a of the IGFBP-2 of by the of IGFBP-2 in of the of IGFBP-2 the of the by the the expression of IGFBP-2 and of and the be a of in cell and IGFBP-2 the of by of and the of in and of of and by and a of and the of to the of of of IGFBP-2 in of in the GBM in and cell the and the in of the cell by the in growth the of growth by IGFBP-2 in cell in IGFBP-2 of by IGFBP-2 in cell of IGFBP-2 of GBM and of IGFBP-2 of and of the IGFBP-2 gene in a of IGFBP-2 in in and growth to of and cell by IGFBP-2 of and by of the of IGFBP-2 in of GBM is a a cell of IGFBP-2 in cell of the and However, of the the the it is to the by the of growth in We a of the of IGFBP-2 in a of the in GBM cell of IGFBP-2 and of GBM. of in of IGFBP-2 IGFBP-2 in of GBM in the the of IGFBP-2 in the growth of in in to IGFBP-2 gene by in in of the IGFBP-2 a of of IGFBP-2 in in expression of IGFBP-2 in by of IGFBP-2 in the in of IGFBP-2 in of and of in the of the of the the of IGFBP-2 the of the expression of IGFBP-2 in the tumor in of the of of by of of IGFBP-2 in expression of GBM and to the in cell IGFBP-2 IGFBP-2 expression and in and to by the by IGFBP-2 in GBM cell has been in and progression of a of of IGFBP-2 expression and expression by We expression of in human GBM and cell GBM by of IGFBP-2 in GBM cell growth of in a by of IGFBP-2 in GBM cell cell in in a IGFBP-2 and of IGFBP-2 in of the the expression of in GBM and cell in GBM and expression of IGFBP-2 and in of GBM cell of is and of in and of of the IGFBP-2 in and of GBM cell expression of IGFBP-2 and However, expression of in and the in cell and cell IGFBP-2 and of the IGFBP-2 of the in the of the the of IGFBP-2 and by and by the in to IGFBP-2 in and be involved in the growth of the GBM is in and is in cell and the is a tumor of and of IGFBP-2 the of of be of the of the of the gene the of by of IGFBP-2 in and and the in of the IGFBP-2 gene by of IGFBP-2 by the IGFBP-2 expression in in of of the of the in in IGFBP-2 the expression of in and of IGFBP-2 the of the in the and by the of IGFBP-2 in of of IGFBP-2 in of IGFBP-2 in in of by the and of of and of of IGFBP-2 and the expression of and the IGFBP-2 and the of IGFBP-2 expression two GBM cell and IGFBP-2 of in in of IGFBP-2 by by the of IGFBP-2 in expression of in cell expression of IGFBP-2 expression in the of in the the of cell of in of GBM the of in of GBM the of of GBM and in the of IGFBP-2 by in a of expression in GBM invasiveness. 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expression in IGFBP-2 GBM a IGFBP-2 and be of the IGFBP-2 gene the growth of GBM in and in IGFBP-2 is in the malignant of GBM be of the by and a in by GBM and the IGFBP-2 and be to the of growth of GBM the the GBM cell in the in GBM in in to the of of the GBM cell in of to the IGFBP-2 and be by GBM the of brain Glioblastoma multiforme (GBM) insulin-like growth factor-binding insulin-like growth short insulin-like growth factor-binding insulin-like growth short is the malignant and is characterized by rapid extensive and extensive of the tumor be by is the of the of GBM remains and the is to the of GBM and to of and of GBM of the of GBM and of growth factor-binding a of the of insulin-like growth in the been been to the a of be to tumor progression IGFBP-2 is a is in the expression expression of IGFBP-2 has been reported in and IGFBP-2 in malignant in expression of IGFBP-2 tumor and IGFBP-2 by GBM by However, it remains to be determined how IGFBP-2 is involved in the growth and of tumor 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the and However, of the the the it is to the by the of growth in We a of the of IGFBP-2 in a of the in GBM cell of IGFBP-2 and of GBM. of in of IGFBP-2 IGFBP-2 in of GBM in the the of IGFBP-2 in the growth of in in to IGFBP-2 gene by in in of the IGFBP-2 a of of IGFBP-2 in in expression of IGFBP-2 in by of IGFBP-2 in the in of IGFBP-2 in of and of in the of the of the the of IGFBP-2 the of the expression of IGFBP-2 in the tumor in of the of of by of of IGFBP-2 in expression of GBM and to the in cell IGFBP-2 IGFBP-2 expression and in and to by the by IGFBP-2 in GBM cell has been in and progression of a of of IGFBP-2 expression and expression by We expression of in human GBM and cell GBM by of IGFBP-2 in GBM cell growth of in a by of IGFBP-2 in GBM cell cell in in a IGFBP-2 and of IGFBP-2 in of the the expression of in GBM and cell in GBM and expression of IGFBP-2 and in of GBM cell of is and of in and of of the IGFBP-2 in and of GBM cell expression of IGFBP-2 and However, expression of in and the in cell and cell IGFBP-2 and of the IGFBP-2 of the in the of the the of IGFBP-2 and by and by the in to IGFBP-2 in and be involved in the growth of the GBM is in and is in cell and the is a tumor of and of IGFBP-2 the of of be of the of the of the gene the of by of IGFBP-2 in and and the in of the IGFBP-2 gene by of IGFBP-2 by the IGFBP-2 expression in in of of the of the in in IGFBP-2 the expression of in and of IGFBP-2 the of the in the and by the of IGFBP-2 in of of IGFBP-2 in of IGFBP-2 in in of by the and of of and of of IGFBP-2 and the expression of and the IGFBP-2 and the of IGFBP-2 expression two GBM cell and IGFBP-2 of in in of IGFBP-2 by by the of IGFBP-2 in expression of in cell expression of IGFBP-2 expression in the of in the the of cell of in of GBM the of in of GBM the of of GBM and in the of IGFBP-2 by in a of expression in GBM invasiveness. However, the of in GBM by of the of the in and cell the by of IGFBP-2 be in by of of of by of the expression in GBM of expression of and of of of the of and of of GBM and in the the of of the gene of GBM in and of in in to the of the of and in the of of the of a of expression IGFBP-2 and of IGFBP-2 and in the expression of and IGFBP-2 in human GBM and GBM cell and brain and in GBM and of the cell and in cell expression of IGFBP-2 in brain the in GBM in brain IGFBP-2 in of human GBM cell expression in of GBM of IGFBP-2 the of IGFBP-2 in GBM cell IGFBP-2 retroviral vectors of a the IGFBP-2 of and IGFBP-2 of two human GBM cell and the of retroviral of expression by a IGFBP-2 in a in the IGFBP-2 in IGFBP-2 the of IGFBP-2 in the expression of IGFBP-2 a We of the of IGFBP-2 in GBM. of by the of it in and the of by the of IGFBP-2 is the IGFBP-2 the of to the the IGFBP-2 gene a of the IGFBP-2 of by the of IGFBP-2 in of the of IGFBP-2 the of the by the the expression of IGFBP-2 and of and the be a of in cell and IGFBP-2 the of of of IGFBP-2 in of in the GBM in and cell the and the in of the cell by the in growth the of growth by IGFBP-2 in cell in IGFBP-2 of by IGFBP-2 in cell and by of the of IGFBP-2 in of GBM is a a cell of IGFBP-2 in cell of the and However, of the the the it is to the by the of growth in We a of the of IGFBP-2 in a of the in GBM cell the of IGFBP-2 in the growth of in in to IGFBP-2 gene by in in of the IGFBP-2 a of of IGFBP-2 in in expression of IGFBP-2 in by of by of of IGFBP-2 in expression of GBM and to the in cell IGFBP-2 IGFBP-2 expression and in and to by the by IGFBP-2 in GBM cell has been in and progression of a of of IGFBP-2 expression and expression by We expression of in human GBM and cell GBM of of GBM cell expression of IGFBP-2 and However, expression of in and the in cell and cell IGFBP-2 and of the IGFBP-2 of the in the of the the of IGFBP-2 and by and by the in to IGFBP-2 in and be involved in the growth of the GBM is in and is in cell and the is a tumor of and of IGFBP-2 the of of be of the of the of the gene the of by of IGFBP-2 in and and the in of the IGFBP-2 gene by of IGFBP-2 by the IGFBP-2 expression in in of of the of the in in IGFBP-2 the expression of in and the IGFBP-2 and the of IGFBP-2 expression two GBM cell and IGFBP-2 of in in of IGFBP-2 by by the of IGFBP-2 in expression of in cell expression of IGFBP-2 expression in the of in the the of cell of in of GBM the of in of GBM the of of GBM and in the of IGFBP-2 by in a of expression in GBM invasiveness. However, the of in GBM by of the of the in and cell the by of IGFBP-2 be in by of to the of the of and in the of of the of a of expression IGFBP-2 and the the of IGFBP-2 in human GBM by of gene retroviral vectors of a the IGFBP-2 of and two human GBM cell and of IGFBP-2 to of the of IGFBP-2 in GBM in and in of IGFBP-2 and of GBM to be in a by of IGFBP-2 to of GBM. IGFBP-2 to a of and of in in IGFBP-2 expression in IGFBP-2 be involved in the growth of GBM the to utilized of the of gene expression of GBM expression of IGFBP-2 expression of IGFBP-2 expression of in GBM and the gene gene expression gene to be by IGFBP-2 in GBM We the is to by GBM expression in two GBM cell and in IGFBP-2 in expression to the to IGFBP-2 expression of IGFBP-2 of of expression by of a expression in GBM in the of of in GBM cell in invasiveness. GBM IGFBP-2 and and of human GBM cell GBM cell and and in and expression of IGFBP-2 be has and by We the of is of the gene in of the of the in to be in the IGFBP-2 in of of and the expression of in in invasiveness. and a and GBM cell IGFBP-2 and is a the of a cell a IGFBP-2 is a cell a in be involved in the a is in a of and is a and be involved in and of brain expression of has been reported in the malignant brain tumor of expression of in a cell the in of is involved in progression and of the the is human GBM and it is involved in the of GBM the of the gene the of the is how IGFBP-2 the IGFBP-2 and remains to be is IGFBP-2 by and of IGFBP-2 in of of However, the of IGFBP-2 the in tumor progression of IGFBP-2 and of GBM the and in GBM IGFBP-2 be by the of IGFBP-2 and in GBM of the of IGFBP-2 of in and to the of the GBM and the a involved in a of by GBM IGFBP-2 the of IGFBP-2 and the expression of in expression the of a by the a in the expression of in IGFBP-2 by and is in of in of expression in IGFBP-2 GBM a IGFBP-2 and be of the IGFBP-2 gene the growth of GBM in and in IGFBP-2 is in the malignant of GBM be of the by and a in by GBM and the IGFBP-2 and be to the of growth of GBM the the GBM cell in the in GBM in in to the of of the GBM cell in of to the IGFBP-2 and be by GBM the the of IGFBP-2 in human GBM by of gene retroviral vectors of a the IGFBP-2 of and two human GBM cell and of IGFBP-2 to of the of IGFBP-2 in GBM in and in of IGFBP-2 and of GBM to be in a by of IGFBP-2 to of GBM. IGFBP-2 to a of and of in in IGFBP-2 expression in IGFBP-2 be involved in the growth of GBM the to utilized of the of gene expression of GBM expression of IGFBP-2 expression of IGFBP-2 expression of in GBM and the gene gene expression gene to be by IGFBP-2 in GBM We the is to by GBM expression in two GBM cell and in IGFBP-2 in expression to the to IGFBP-2 expression of IGFBP-2 of of expression by of a expression in GBM in the of of in GBM cell in invasiveness. GBM IGFBP-2 and and of human GBM cell GBM cell and and in and expression of IGFBP-2 be has and by We the of is of the gene in of the of the in to be in the IGFBP-2 in of of and the expression of in in invasiveness. and a and GBM cell IGFBP-2 and is a the of a cell a IGFBP-2 is a cell a in be involved in the a is in a of and is a and be involved in and of brain expression of has been reported in the malignant brain tumor of expression of in a cell the in of is involved in progression and of the the is human GBM and it is involved in the of GBM the of the gene the of the is how IGFBP-2 the IGFBP-2 and remains to be is IGFBP-2 by and of IGFBP-2 in of of However, the of IGFBP-2 the in tumor progression of IGFBP-2 and of GBM the and in GBM IGFBP-2 be by the of IGFBP-2 and in GBM of the of IGFBP-2 of in and to the of the GBM and the a involved in a of by GBM IGFBP-2 the of IGFBP-2 and the expression of in expression the of a by the a in the expression of in IGFBP-2 by and is in of in of expression in IGFBP-2 GBM a IGFBP-2 and be of the IGFBP-2 gene the growth of GBM in and in IGFBP-2 is in the malignant of GBM be of the by and a in by GBM and the IGFBP-2 and be to the of growth of GBM the the GBM cell in the in GBM in in to the of of the GBM cell in of to the IGFBP-2 and be by GBM We to and