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Comments:
The RKO-E6 cell line was generated from the colon carcinoma RKO (ATCC CRL-2577) cell line by transfection with pCMV-E6 using Lipofectin. [53390]
The cells contain a stably integrated human papilloma virus (HPV) E6 oncogene under control of the cytomegalovirus (CMV) promoter. The HPV E6 oncogene causes a decrease in normal p53 levels and functions.
The RKO-E6 cell line lacks appreciable functional p53.
Disruption of normal p53 function in human colon carcinoma RKO cells with the human papillomavirus E6 oncoprotein results in reduced repair of u.v.-induced DNA damage and also loss of induced repair following cellular u.v.-irradiation. [51463]
RKO cells contain wild-type p53 but lack endogenous human thyroid receptor nuclear receptor (h-TRbeta1). The level of p53 protein is higher in RKO (ATCC CRL-2577) cells than in RKO-E6 (ATCC CRL-2578) cells. [53331]
The RKO-E6 cell line can be used together with its parental cell line, RKO (ATCC CRL-2577) to investigate the effects of p53 loss on cellular parameters such as p53 mediated transcription and apoptosis.
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 1:4 to 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:
51463: Smith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629 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 53331: Bhat MK, et al. Tumor suppressor p53 is a negative regulator in thyroid hormone receptor signaling pathways. J. Biol. Chem. 272: 28989-28993, 1997. PubMed: 9360971 53390: Kessis TD, et al. Human papillomavirus 16 E6 expression disrupts the p53-mediated cellular response to DNA damage. Proc. Natl. Acad. Sci. USA 90: 3988-3992, 1993. PubMed: 8387205
