Background
Results
Imaging of the tumor before surgery
Surgical therapy effectively debulked the tumor mass
Pathological analysis determined high-grade glioma
Tumor grew back after surgical and adjuvant therapies as monitored by CT and MRI
Cultured CD133-expressing cells behaved as cancer cells
Diverse marker expression
Stem cell markers
|
Proliferation, angliogenic, migration markers, growth factors, adhesion
| ||
---|---|---|---|
SOX2 | ++++++ | Ki67 | +++++ |
DACH1 | + | VEGF | +++ |
CD133 | ++++ | HIF1 | + |
Nestin | +++ | CXCR4 | ++ |
Nucleostemin | ++++ | SDF1 | + |
IGF1R | ++ | ||
NCAM | +++ | ||
Glial Cells
| PAX6 | ++++ | |
GFAP | ++ | ||
Ephrins receptors and ligands (cell repulsion, adhesion, migration)
| |||
Neuronal
| EphB1 Receptor | n/a | |
DCX | + | EphB2 Receptors | ++++++ |
MSI | + | EphB3 Receptors | +++++ |
MAP2 | + | EFNB1\ | + |
Neural Specific Endolase | ++++ | EFNB2 | ++ |
Beta-3Tubulin III | + | EFNB3 | + |
MMPs, Peptidases
| ++ | ||
MMP2 | +++ | ||
Tumor Suppressors/Oncogenes
| TIMP1 | ||
p53 | + | MMP9 | +++ |
PTEN | + | MMP13 | ++++ |
Bmi-1 | ++ | MMP14 | + |
CAV-1 | ++ | Cathepsin B | + |
CAV-2 | + | Cathepsin L | +++ |
c-Myc | +++ | ||
Controls
| |||
Proteoglycans
| GAPDH | +++ | |
Mucin9 | ++ | Beta-Actin | ++ |
Mucin18 | + | H2O | |
Mucin24 | ++++ | ||
Notch-Family
| |||
Notch1 | + | ||
Notch2 | +++ | ||
Notch3 | + | ||
Notch4 | ++ |
Caveolin-1 is expressed in the CD133-positive cells
CD133-positive cells formed brain tumors in vivo
Discussion
Conclusions
Methods
Patient’s background
Surgery
Tumor histology
Tumor cell culture
Clonogenic assay
Preparation of organotypic slices from murine brain tissue
Transplantation of cells onto organotypic brain slices
Semi-quantitative RT-PCR
Gene | Sequences (5' to 3') |
---|---|
Beta-Actin F
| GCACCACACCTTCTACAATGAGC |
Beta-Actin R
| TTGAAGGTAGTTTCGTGGATGCC |
Beta-tubulin III F
| AACGAGGCGCTCTACGACATC |
Beta-tubulin III R
| CTCCTCCTCGTCGTCTTCGTA |
Bmi-1AS
| CATTGCTGCTGGGCATCGTAAG |
Bmi-1 S
| GGAGACCAGCAAGTATTGTCCTTTTG |
BMP4 F
| GTGAGGAGCTTCCACCACGA |
BMP4 R
| ACTGGTCCCTGGGATGTTCTC |
Cathepsin B F
| GCAGCCTCAGCCACCCAGAT |
Cathepsin B R
| CCACCATTACAGCCGTCCCCACAC |
Cathepsin L F
| CCGGGGAGGGCAGTTGAG |
Cathepsin L R
| CCTTGAGGCCCAGAGCAGTC |
Cav-1 F
| GGACATCTCTACACCGTTCCC |
Cav-1 R
| TTATATTTCTTTCTGCAAGTT |
Cav-2 F
| ATGGGGGCTGGAGACGGAGAAG |
Cav-2 R
| TCAATCCTGGCTCAGTTGCAG |
CD133 AS
| ACGCCTTGTCCTTGGTAGTGTTG |
CD133 S
| CTGGGGCTGCTGTTTATTATTCTG |
c-Myc F
| AAGGTCAGAGTCTGGATCAC |
c-Myc R
| TAACTACCTTGGGGGCCTTT |
CXCR4 F
| CACCGCATCTGGAGAACCA |
CXCR4 R
| GCCCATTTCCTCGGTGTAGTT |
Dach1 S
| AGGCTTTCGACCTGTTCCTGAA |
Dach1 AS
| GCTGTCAGACCTGTTGGTGGAA |
DCX AS
| GTTTCCCTTCATGACTCGGCA |
DCX S
| AATCCCAACTGGTCTGTCAAC |
EPHB1 S
| GAGATGGACAGCTCCAGAGG |
EPHB1 AS
| CCAGCATGAGCTGGTGTAGA |
EPHB2 S
| AAAATTGAGCAGGTGATCGG |
EPHB2 AS
| TCACAGGTGTGCTCTTGGTC |
EPHB3 S
| AGCAACCTGGTCTGCAAAGT |
EPHB3 AS
| TCCATAGCTCATGACCTCCC |
EFNB1 S
| GGAGGCAGACAACACTGTCA |
EFNB1 AS
| GAACAATGCCACCTTGGAGT |
EFNB2 S
| GCAAGTT-CTGCTGGATCAAC |
EFNB2 AS
| AGGATG-TTGTTCCCCGAATG |
EFNB3, S
| CTGAAATGCCCATGGAAAGA |
EFNB3, AS
| ACGCCCAGCAAGAGCAGCGC |
GAPDH S
| ACCACAGTCCATGCCATCAC |
GAPDH AS
| TCCA CCACCCTGTTGCTGTA |
GFAP S
| ACATCGAGATCGCCACCTAC |
GFAP AS
| ACATCACATCCTTGTGCTCC |
HIF1 S
| GTCGGACAGCCTCACCAAACAGAGC |
HIF1 AS
| GTTAACTTGATCCAAAGCTCTGAG |
IGF1R S
| ACGCCAATAAGTTCGTCCAC |
IGF1R AS
| TCCATCCTTGAGGGACTCAG |
Ki67 F
| GGAGGCAATATTACATAATTTCA |
Ki67 R
| CAGGGTCAGAAGAGAAGCTA |
MMP2 F
| CCA CGT GAC AAG CCC ATG GGG CCC C |
MMP2 R
| GCA GCC TAG CCA GTC GGA TTT GAT G |
MMP9 F
| CAACATCACCTATTGGATCC |
MMP9 R
| CGGGTGTAGAGTCTCTCGCT |
MMP13 F
| TGCTCGCATTCTCCTTCAGGA |
MMP13 R
| ATGCATCCAGGGGTCCTGGC |
MMP14 F
| CGCTACGCCATCCAGGGTCTCAAA |
MMP14 R
| CGGTCATCATCGGGCAGCACAAAA |
msi1 S
| GAGACTGACGCGCCCCAGCC |
msi1 AS
| CGCCTGGTCCATGAAAGTGACG |
Mucin9 F
| CATGACCGGTGGACTTTTCT |
Mucin9 R
| TCCTGTGAACCTTTCCCAAC |
Mucin18 F
| GTGTTGAATCTGTCTTGTGAA |
Mucin18 R
| ATGCCTCAGATCGATG |
Mucin24 F2
| GTTAATACTACCTGCTTTTGGATAGAATGT |
Mucin24 R2
| CCACTTGACAATCACTAACTGTTGAG |
NCAM1 AS
| GGTGTTGGAAATGCTCTGGT |
NCAM1 S
| AGGAGACAGAAACGAAGCCA |
Nestin
| GGCAGCGTTGGAACAGAGGTTGGA |
Nestin R
| CTCTAAACTGGAGTGGTCAGGGCT |
Notch1 F
| TGTTAATGAGTGCATCTCCAA |
Notch1 R
| CATTCGTAGCCATCAATCTTGTCC |
NSE AS
| GACAGTTG CAGGCCTTTTCTTC |
NSE S
| CATCGA CAAGGCTGGCTACACG |
Nucleostamin F
| CATGACCTGCCATAAGCGGT |
Nucleostamin R
| CAATTACTCCAACCCGAATGGC |
Pax6 F
| GAGCCTCATCTGAATCTTCTCCG |
Pax6 R
| CGTCCATCTTTGCTTGGGAAATC |
p53 S
| TTGGATCCATGTTTTGCCAACTGGCC |
p53 AS
| TTGAATTCAGGCTCCCCTTTCTTGCG |
PTEN F
| GGACGAACTGGTGTAATGATATG |
PTEN R
| TCTACTGTTTTTGTGAAGTACAGC |
SDF-1S
| GGGGGAATTCCATGAACGCCAAGGTCGTGGTC |
SDF-1 AS
| GGGGTCTAGAGGGCATGGATGAATATAAGCTGC |
Sox2 S
| ACCGGCGGCAACCAGAAGAACAG |
Sox2 AS
| GCGCCGCGGCCGGTATTTAT |
TIMP1 F
| ACT GGA AGC CCT TTT CAG AGC |
TIMP1 R
| AAT TCC GAC CTC GTC ATC AGG |
VEGF AS
| CGATCGTTCTGTATCAGTCTTTCC |
VEGF S
| GAAGTGGTGAAGTTCATGGATGTC |