synthesis according to the manufacturer’s protocol. Real-time PCR reactions were then carried out for a total of 25 L reaction mixture SmartCycler II. The PCR program was 10 min at 95 40 initiated from thermal WYE-354 cycles of 15 s at 95 min and 1 to 60. The data were calculated using the comparative Ct method and were normalized by actin expression in each sample. Melting curves were generated for each PCR reaction to hrleisten weight purity of the amplification. Western blot PC 3 cells were sown in bo t Your 100 mm culture 3.0 105 cells per bo And allow yourself to attach for 24 hours overnight. The cells were treated with 10 nM SB715992 for 24 and 48 hours. The cells were then lysed in 62.5 mM Tris-HCl and 2 SDS. Protein concentrations of each sample was.
Using a BCA protein assay kit Cell protein extracts were then subjected to 10 or 14 of the SDS-PAGE and transferred to nitrocellulose membranes at 100 V for 2 hours 4 Membranes were incubated with anti-EGFR antique Body, anti p27, p15 and anti-anti-actin prim Ren antique Rpern incubated and then End with secondary Ren antique Rpern conjugated to peroxidase. The signal was then determined using A-966492 chemiluminescence detection system. Test for statistical analysis of cell growth inhibition and apoptosis ELISA was statistical analysis using the t-test between treated and untreated samples or between mono and combination therapy treatment. P values of less than 0.05 indicate statistical significance.
Inhibition of PC 3 Results cell proliferation of prostate cancer cells by SB715992 PC 3 prostate cancer were treated with 15 and 30 nM SB715992 and regarding inhibition of cell proliferation. The results of the MTT assay showed that a time and dose-SB715992-Dependent effect on the growth of the PC had 3 cells. Within 72 hours of an average inhibition of cell growth of SB715992 and 48.65 52.16 15 nm to 30 nm in comparison with control samples caused. Therefore, the results of this experiment that SB715992 is a potent inhibitor of cell growth in vitro, PC 3. With this information, we conducted our study on the F Ability, induce apoptosis in SB715992 PC 3 prostate cancer cells. Induction of apoptosis in prostate cancer cells by SB715992 The induction of apoptosis was tested by two different methods. First, the induction of apoptosis was detected by SB715992 by ELISA.
The results of the ELISA were obtained showed an average of 1094.88 increase in apoptosis in PC 3 cells with 15 nM SB715992 treated and 1516.70, when treated with 30 nM SB715992 for 72 hours. As a result, showed an effect of SB715992 time and dose-dependent-Dependent increase in apoptosis, which correlates with the results obtained by MTT assay. Therefore, we have decided to take our data by ELISA analysis of DNA ladder at best Term. The results of the DNA ladder analysis also showed a Erh Increase in time with SB715992 induces apoptosis through assigned. Thus, the results of both ELISA and DNA ladder are analysis clearly showed that SB715992 was a potent inducer of apoptosis in prostate cancer cells. Consequently, the information from the analysis of apoptosis and inhibition of cell proliferation resulting invited u