A Radiation-Induced Acute Apoptosis Involving TP53 and BAX Precedes the Delayed Apoptosis and Neoplastic Transformation of CGL1 Human Hybrid Cells

DMAPT inhibits NF-κB activity and increases sensitivity of prostate cancer cells to X-rays in vitro and in tumor xenografts in vivo

“…Twenty-four and forty-eight hours after plating, appropriate volumes of 36 mM DMAPT drug stock diluted in RPMI were directly added to the cell culture flasks so that final DMAPT concentrations were 0, 2.5, and 5 μM DMAPT for PC-3 and 0 and 4 μM DMAPT for DU145 cells. Control and treated cells were harvested for NF-κB EMSA/Gel shifts, cell counting/growth curve analysis, clonogenic potential/plating efficiency studies, flow cytometry based apoptosis analysis by Annexin V-phosphatidylserine analysis, comet assays to determine DNA double strand break repair, flow cytometry based DNA histogram cell cycle analysis after propidium iodide staining, and Western analyses 24 and 48 h after drug addition by our standard techniques [12,26,28,45,46]. Since the initial NF-κB EMSA/Gel shifts clearly showed better inhibition of NF-kB binding at 5 μM DMAPT, all other experiments with PC-3 were performed at 5 μM DMAPT.…”

“…Twenty-four and forty-eight hours after treatment of PC-3 and DU145 cells with 0 or 5 μM of DMAPT, or 0 or 4 μM of DMAPT, respectively, cells were analyzed for changes in protein expression by our standard Western analysis [26,28,46]. The primary antibodies utilized were anti-p53 (Ab-6, Calbiochem and clone BP53–12 Upstate), anti-p73 (rabbit polyclonal, Chemicon), anti-p21 waf−1/Cip−1 (clone 6B6, BD Pharmingen and mixed monoclonal, Upstate), anti-Bax (Clone YTH-2D2, Trevigen), anti-Bcl-2 (clone 124, Dako), anti-Bid (BD Pharmingen), and anti-α-tubulin (cloneB-5–1–2, Sigma).…”

Role of p53 in Regulating Radiation Responses

“…We have reported that changes in T-cell surface markers occur in a delayed manner when mice are exposed to whole body irradiation, and this phenomenon was observed earlier in p53 heterozygotes [ 122 ]. X-irradiated cells undergo delayed apoptosis, which may not be a result of nonfunctioning of the acute TP53 damage response pathway but rather X-ray-induced genomic instability that occurs in the remote descendants of irradiated cells [ 123 ].…”

Substance Change in Pro-Apoptosis and Pre- Apoptosis in Malignant Transformation of Cell Clusters in Carcinogenesis

“…It is further to such molecular binding to receptors and decoy receptors that a concept of molecular dynamics is a reflected dimension of turnover in terms that predispose to conformational modification and to post-translational change in the binding of interface modalities. Substantial attributes of dysfunction are further accentuated within predisposed susceptibilities to a state of enhanced apoptotic susceptibility, as terms that additionally require the outline emergence for further progressive cascades of interactivity, A radiation-induced acute apoptosis involving TP53 and BAX precede delayed apoptosis and malignant transformation of CGL1 human hybrid cells derived from (HeLa X fibroblast) [13].…”