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Oncogene:
p53 + (underexpressed)
Comments:
The RKO-AS45-1 cell line was generated from the colon carcinoma RKO (ATCC CRL-2577) cell line by transfection with the open reading frame portions of human cDNAs for GADD45 cloned into the expression vector pCMV. [51464]
The cells contain a stably integrated cytomegalovirus (CMV) promotor driven expression vector. The cells overexpress Gadd45 RNA and protein.
The GADD45 gene is p53-regulated and the Gadd45 protein interacts with proteins involved in both cell-cycle regulation and DNA repair.
The RKO-AS45-1 cell line has reduced ability to repair U.V.-induced DNA damage. [51464]
The RKO-AS45-1 cell line can be used together with its parental cell line, RKO (ATCC CRL-2577) to investigate the effects of Gadd45 on DNA repair, apoptosis and drug sensitivity.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C
Subculturing:
Subcultivation Ratio: A subcultivation ratio of 1:10 is recommended Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Preservation:
culture medium 95%; DMSO, 5%
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003
recommended serum:ATCC 30-2020
parental cell line:ATCC CRL-2577
References:
51464: Smith ML, et al. Antisense GADD45 expression results in decreased DNA repair and sensitizes cells to u.v.-irradiation or cisplatin. Oncogene 13: 2255-2263, 1996. PubMed: 8950993 53491: Smith ML, et al. Interaction of the p53-regulated protein Gadd45 with proliferating cell nuclear antigen. Science 266: 1376-1380, 1994. PubMed: 7973727 53492: Bae I, et al. An abnormality in the p53 pathway following gamma-irradiation in many wild-type p53 human melanoma lines. Cancer Res. 56: 840-847, 1996. PubMed: 8631022
