Table 2.
Oligonucleotide primers used for RT-PCR of cyclin D1, GST-P, VEGF, VEGFR1, MMP-9, TIMP-2 with β-actin as an internal control
| Gene Product | Primers | Oligonucleotide sequences | Fragment size |
|---|---|---|---|
|
| |||
| Cyclin D1 | Sense | 5'-CGGAGGACAACAAACAGATC-3' | 331 bp |
| Antisense | 5'-GGGTGTGCAAGCCAGGTCCA-3' | ||
|
| |||
| GST-P | Sense | 5'-TCATCTACACCAACTATGAG-3' | 226 bp |
| Antisense | 5'-GCCACATAGGCAG AGAGCAG-3' | ||
|
| |||
| VEGF | Sense | 5'-ATGAACTTTCTGCTGTCTTGG-3' | 444 bp and 576 bp* |
| Antisense | 5'-TCACCGCCTCGGCTTGTCACA-3' | ||
|
| |||
| VEGFR1 | Sense | 5'-AGGAGAGGACCTGAAACTGTCTT-3' | 230bp |
| Antisense | 5'-ATTCCTGGGCTCTGCAGGCATAG-3' | ||
|
| |||
| MMP-9 | Sense | 5'-AGTTTGGTGTCGCGGAGCAC-3' | 753bp |
| Antisense | 5'-TACATGAGCGCTTCCGGCAC-3' | ||
|
| |||
| TIMP-2 | Sense | 5'-GTTTTGCAATGCAGACGTAG-3' | 539bp |
| Antisense | 5'-ATGTCAAGAAACTCCTGCTT-3' | ||
|
| |||
| β-actin | Sense | 5'-AACCGCGAGAAGATGACCCAGATCATGTTT-3' | 350 bp |
| Antisense | 5'-AGCAGCCGTGGCCATC TCTTGCTCGAAGTC-3' | ||
*
The primers for VEGF detect two of four different molecular species produced by alternative splicing of mRNA-VEGFI21 and VEGFI65, with expected fragment sizes of 444 bp and 576 bp, respectively.