JunB gene expression levels and methylated CpG frequencies at the promoter area in healthy individuals and patients with CML
| . | No. . | Real-time quantitative RT-PCR . | Methylation study . | ||||
|---|---|---|---|---|---|---|---|
| ΔCT. JunB-GAPDH . | ΔΔCT. CML-normal . | 2−ΔΔCT . | Methylation rate . | No. of CpG . | No. of T/Cp G . | ||
| Healthy | 17 | 0.98 ± 0.32* | 0/17 (0%) | 0 | 0 | ||
| CML | |||||||
| Blastic crisis | 11 | 5.94 ± 0.78† | 4.96 | 1/31.12 | 11/11 (100%) | 12.64 ± 0.31 | 3.36 ± 0.31 |
| Chronic phase | 21 | 3.22 ± 0.47† | 2.24 | 1/4.72 | 21/21 (100%) | 12.50 ± 0.39 | 3.50 ± 0.39 |
| . | No. . | Real-time quantitative RT-PCR . | Methylation study . | ||||
|---|---|---|---|---|---|---|---|
| ΔCT. JunB-GAPDH . | ΔΔCT. CML-normal . | 2−ΔΔCT . | Methylation rate . | No. of CpG . | No. of T/Cp G . | ||
| Healthy | 17 | 0.98 ± 0.32* | 0/17 (0%) | 0 | 0 | ||
| CML | |||||||
| Blastic crisis | 11 | 5.94 ± 0.78† | 4.96 | 1/31.12 | 11/11 (100%) | 12.64 ± 0.31 | 3.36 ± 0.31 |
| Chronic phase | 21 | 3.22 ± 0.47† | 2.24 | 1/4.72 | 21/21 (100%) | 12.50 ± 0.39 | 3.50 ± 0.39 |
JunB gene expression was determined by real-time quantitative RT-PCR. The amount of JunB was normalized to the endogenous reference GAPDH to obtain the ΔCT value for each sample. The normalized JunB expression (ΔCT) of CML cases was first related to the ΔCT of normal cases to obtain the relative threshold cycle (ΔΔCT) and then the relative expression levels (2−ΔΔCT) were calculated. In the methylation study, all the healthy cases are positive for the nonmethylation-specific PCR and all the CML cases are positive for methylation-specific PCR. The numbers of CpG and T/Cp G were calculated from direct sequencing results of JunB-U or JunB-M PCR products of bisulfite-modified DNA.
Results are the means ± SEs.
P < .001 when compared with normal.