Table 2.
Nuclear and Cytoplasmic Staining of Human Cell Lines for FANCD2
| Breast cell line* | Classification* | Percentage stained†
|
|
|---|---|---|---|
| Nucleus | Cytoplasm | ||
| SVE3 | Immortalized normal | 23 ± 5 | 53 ± 12 |
| Huma 7 | Immortalized normal | 23 ± 4 | 52 ± 13 |
| Huma 121 | Immortalized benign | 24 ± 5 | 43 ± 8 |
| Huma 123 | Immortalized benign | 21 ± 3 | 53 ± 4 |
| MCF-7 | Malignant: low invasive | 8 ± 3§ | 53 ± 8 |
| T47D | Malignant: low invasive | 5 ± 2§ | 51 ± 6 |
| MDA-MB-157 | Malignant: high invasive | 0 ± 0§ | 1 ± 1§ |
| MDA-MB-231 | Malignant: high invasive | 0 ± 0§ | 1 ± 1§ |
| FANCD2-deficient cell line‡ | Mitomycin C‡ | Percentage stained†
|
|
| Nucleus
|
Cytoplasm
|
||
| PD20 | − | 2 ± 1 | 2 ± 1 |
| + | 2 ± 1 | 2 ± 1 | |
| PD733 | − | 2 ± 1 | 3 ± 1 |
| PD20-3-15 | − | 11 ± 4¶ | 63 ± 1¶ |
| + | 71 ± 11¶ | 32 ± 8¶ | |
Epithelial cell lines were derived from normal (SVE3, Huma 7), benign (Huma 121, Huma 123), malignant low invasive (MCF-7, T47D), or malignant high invasive (MDA-MB-157, 231) breast sources (Materials and Methods).
Immunocytochemical staining for FANCD2 showing the mean percentage ± standard deviation (n = 8) of positive nuclei and cytoplasms.
PD20 (FANCD2-deficient) or PD20-3-15 (PD20 functionally complemented by microcell mediated transfer of chromosome 3p) fibroblasts were grown in the presence (+) or absence (−) of 50 nmol/L mitomycin C for 18 hours. A second unrelated FANCD2-deficient fibroblast cell line PD733 was grown without MMC.
Significantly different in Student’s t-test for nuclear staining for SVE3, Huma 7, Huma 121, Huma 123, compared to MCF-7, T47D, MDA-MB-157, MDA-MB-231 (P < 0.0001) and in cytoplasmic staining for SVE3, Huma 7, Huma 121, Huma 123, MCF-7, ZR-75 compared to MDA-MB-157, MDA-MB-231 (P < 0.0001) and for MCF-7, T47D versus MDA-MB-157, MDA-MB-231 for nuclear and cytoplasmic staining (P ≤ 0.0002).
Significantly different in Student’s t-test for PD20 vs PD20-3-15 (P ≤ 0.0003) and for PD20-3-15 with (+) MMC versus without (−) MMC for nuclear and cytoplasmic staining (P < 0.0001) but not for PD20 vector alone with (+) 50 nmol/L MMC versus without (−) MMC for nuclear and cytoplasmic staining (P = 1).