Androgen Receptor Expression, Proliferation Index and Aneuploidy in Tissue Explant Cultures Derived Prostate Carcinoma Cells Co-Cultivated on Membranes

Pressure regulated basis for gene transcription by delta-cell micro-compliance modeled in silico: Biphenyl, bisphenol and small molecule ligand models of cell contraction-expansion

“…The grade of P eff duration for effect is then determined for the extracellular subset of small molecule hormone nuclear receptor ligands (Ch 4 O nl L external structure /H polar group : 0.461, Dex– 1.31, Des) with molecular diameters within the 0.774 nm (Bpaf)– 0.873 nm (Dex) vdWD range with a minimum of di-polar hydroxylation hydrophilicity (- 2.10 nm -1 ), which is on the basis of small molecule hormone and inverse agonist potential for disassociation over range of exposure concentration (K d ) and binding affinity over time ( t 1/2 ) [ 111 – 115 ] as plotted independent variables for semi-exponential power-regression extrapolation of half-life at receptor of unknowns ( R 2 = 0.955), and applied whole cell receptor density based on magnetic bead-enhanced amphometric detection or radioligand competition assay studies (B max ; n ) [ 115 , 116 ] for multiplicative in silico modeling of pressure regulatory grade of effect for gene expression in a mono-compliant cell type. The half-lives at receptor ( t 1/2 ) for diethylstilbestrol (Des) at ERα is 663 min, and that for dihydrotestosterone (Dht) and methyltrienolone (R1881) at AR are within the 38–53 minutes (min) interval, as the K d approximates 0.9–1.0 nM (see ref 118 ). The strata order for ligand · receptor grade of duration at P eff , from positive with contraction-expansion response-to-negative, is 1.4215E + 04 [Cort · MR (GR)], 3.0270E + 04 [Ald · MR (GR)], 1.33383E + 05 [Dex ~ Corticosterone · GR (MR)], 1.79812E + 05 (Dht · AR), 2.47340E + 05 (R1881 · AR), 1.2E +06 (E 2 · ERα), 1.863745E + 06 (DES · ERα) adjusted for whole cell receptor count ( Σ min · count ), from positive to negative ( Table 9 ).…”

“…Thus, based on study determinations, the gene expression pattern for small molecule hormone receptor interaction between the 2.6 x 10 5 to 2.1 x 10 6 min�count range results in a negative Δ C micro response and an initial downward shift in the contraction with unilateral expansion as compared to positive Δ C micro response with contraction and bidirectional expansion, and irrespective of the specific steriod axis receptor class (ER, AR). Furthermore, it appears that an initial negative (-) Δ C micro response within the 1.86 x 10 6 to 1.94 x 10 6 (IGF-II � IGF-IIR) min�count range is coupled to a positive Δ C micro response, as Dht or R1881 � CM AR (1.79E + 05-2.45E + 05, + P eff ) results in the transcription of pro-proliferative genes [118,119], that is proposed to be by an initial (-) P eff Χ then an (+) P eff Y intermediate step coupled to resultant transcriptional activation of MKI67 (P eff 0.329) [Part I, not cited; 38] ( Table 9); thus, expression of a focal adhesion or endocytic component may be involved, which would apply to calvarial osteoblasts that overexpress the IGF-IIR/M6P receptor [120], and similarly to transformed cells with P eff shift to Several further studies point in the direction of paradoxical responses to dexamethasone (Dex) treatment, as example of a biologic mimic that produces a parabolic peak grade of positive P eff response and corticosterone surrogate, in which case observed divergent gene expression responses upon applied Dex are in dissimilar cell types [123], due to dose response and variant GR receptor affinity [124], or during the tuned activation of Dex responsive genes with GRE sequence sites in proximity to the TSS [125]. The magnitude of differential gene expression response in cultured primary astrocyte and neurons to Dex stimulation is consistent with respective increases in duration at P eff contraction-expansion phase gene expression in a less and more compliant cell type to the same agent (ie PER1, FKBP5; 5.3x) [123], in which case it appears that the difference in magnitude of differential gene expression achieved in-between cell types is unlikely attributable to ligand � receptor min�count, as astrocyte GR mRNA is 3x-overfold neuronal in which case only an apparent difference in t 1/2 at receptor exists; whereas, the same in the high affinity variant porcGR (ala 610 val) transgenic model, in which an under-expression of GCLC (P eff 0.477) and PCK1 (P eff 0.333), and overexpression of FKBP5 (P eff 0.342) follows a saturable dose-escalation differential gene expression pattern [124], and is attributable to an upward contraction-expansion shift in P eff response with maintained range of ligand � receptor affinity.…”

Tumour-stroma interactions between metastatic prostate cancer cells and fibroblasts

“…We used three established metastatic human prostate cancer cell lines on the one hand, and a human fibroblast line on the other which we had well characterized in previous studies (56,58). Cell isolation from fresh prostate tissues is possible (59,60) but has inherent problems such as preparation of 'pure' cell populations and short life-span of primary cells. In addition, the genetic background of different patients renders data difficult to interpret and to compare.…”

Primary Culture of Pancreatic (Human) Acinar Cells

“…The cells were washed twice and incubated with secondary antibody, FITC labelled The quantitative determination of DNA synthesis in acinar cell culture was done by using the cell proliferation BrdU ELISA kit (Roche, Germany). Briefly, 10 4 cells/100 ll were seeded in 96-well plates and labelled with BrdU at final concentration of 1 lM for 24 h [15]. The labelling medium was removed and cells were further incubated with FixDenat solution for 30 min at 4°C.…”