Table 1.
Relation between induction of wip1 and p53 status
| Cell line | Cell type | p53 status* | G1 arrest | Apoptosis | Fold induction of relative mRNA†
|
|
|---|---|---|---|---|---|---|
| wip1 | waf1 | |||||
| Wild-type p53 status and function | ||||||
| ML-1 | Myeloid leukemia | wt/wt | + | + | 5.0 ± 0.8† | 16.2 |
| WMN | Burkitt lymphoma | wt/wt | + | + | 3.2 ± 0.6 | 4.1 ± 0.5 |
| FWL | Lymphoblastoid | wt/wt | + | + | 2.9 ± 0.4 | 6.0 ± 0.4 |
| Shoemaker | Lymphoblastoid | wt/wt | + | + | 2.6 ± 0.2 | 5.6 |
| NL2 | Lymphoblastoid | wt/wt | + | + | 2.5 ± 0.1 | 6.6 |
| H460 | NSC lung carcinoma | wt/wt | + | 2.2 ± 0.5 | 4.5 | |
| MCF-7 | Breast carcinoma | wt/wt | + | +‡ | 2.0 ± 0.1 | 4.5 |
| Abnormal p53 status and function | ||||||
| CA46 | Burkitt lymphoma | mut/−§ | − | − | 1.0 ± 0.0 | 1.7 ± 1.1 |
| JD38 | Burkitt lymphoma | mut/− | − | − | 1.2 ± 0.1 | 2.1 |
| MC116 | Burkitt lymphoma | mut/− | − | − | 1.2 ± 0.1 | 3.9 |
| Ramos | Burkitt lymphoma | mut/− | − | − | 1.1 ± 0.1 | 1.5 |
| Transfected cell lines | ||||||
| HCT116-CMV¶ | Colon carcinoma | wt/wt | + | 2.0 | 3.5 | |
| HCT116-E6¶ | Colon carcinoma | wt/wt (E6) | − | 1.1 | 1.4 | |
| Calu6 | Lung carcinoma | −/− | 0.7** | 1.0 | ||
| Calu6-143‖ | Lung carcinoma | −/− (tsp53) | 2.5 | 8.0 | ||
| Murine cells | ||||||
| MEF | Fibroblasts | +/+ | 2.5 | |||
| MEF | Fibroblasts | −/− | 1.0 | |||
| PNK | Keratinocytes | +/+ | 8.4‡‡ | |||
| PNK | Keratinocytes | +/− | 4.0 | |||
| PNK | Keratinocytes | −/− | 1.2 | |||
Note: When not determined, the postition was left blank.
Data regarding p53 status, G1 arrest, and apoptosis are taken from ref. 26 and references therein.
Human cell lines with normal or abnormal p53 status were irradiated with 6.3 Gy of IR. Relative values for samples extracted 4 hr after treatment, compared with untreated controls, were determined by densitormetric scanning of Northern blots using PhophoImager model 425 (Molecular Dynamics). wip1 and waf1 values are normalized relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Where shown, data are ±SD from mean value of three experiments.
Apoptosis could be detected but was substantially less than for other cell lines (23).
Only one mutant form of p53 was detected, indicating either that both alleles contained the same mutation or that one allele had been deleted (23).
HCT116 cell line was transfected with either pCMVneo vector or with pCMVneo-E6, encoding for the HPV-E6 protein known to abrogate p53-dependent transactivation (27).
Calu6 cells were transfected with a pCMVneo vector encoding for a human temperature-sensitive p53 (p53-V143A) (28), generating Calu6-143. Calu6-143 expresses p53 either in a mutant or wt conformation when cultured at 37°C or 30°C, respectively.
Induction was determined comparing RNA expression from cells cultured 24 hr at 30°C (permissive temperature) to control cultures at 37°C.
Primary newborn keratinocytes (PNK) were treated with 50 J/m2 UVC. Relative values for samples extracted 24 hr after treatment, compared with untreated controls, were determined by densitometric scanning of Northern blots (W. C. Weinberg and E. Fernandez-Salas, personal communication).