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. Author manuscript; available in PMC: 2015 Mar 10.
Published in final edited form as: J Cell Biochem. 2009 Nov 1;108(4):998–1011. doi: 10.1002/jcb.22333

Fig. 1.

Fig. 1

8-CPT upregulates [3H]thymidine uptake, protein synthesis and proliferation in 1-LN prostate cancer cells. Panel A: [3H]Thymidine uptake by 1-LN cells treated with varying concentrations of 8-CPT. [3H]Thymidine uptake is expressed as fmols [3H]thymidine incorporated into DNA/mg protein and is the mean ± SE from two experiments performed in triplicate. The bar diagram on right side of Panel B shows the effect of pretreatment of 1-LN cells with LY294002 or rapamycin on 8-CPT-induced [3H]thymidine uptake by 1-LN cells. The bars are (1) vehicle; (2) 8-CPT (50 μM/16h); (3) LY294002 (20 μM) then 8-CPT; and (4) rapamycin (100 nM) then 8-CPT. [3H]Thymidine uptake is expressed as above as the mean ± SE from two experiments performed in triplicate. Panel C: [3H]Leucine incorporation (pmol/mg protein) into 1-LN cells treated with varying concentrations of 8-CPT. To the right of Panel C, the bar diagram, Panel D, shows the effect of pretreating the cells with inhibitors of farnesyl transferase, MAPKs, PI 3-kinase, or mTOR, respectively in 8-CPT-induced [3H]leucine incorporation (pmol/mg/protein) into cellular protein in 1-LN cells. The bars are: (1) vehicle; (2) 8-CPT (100 μM); (3) manumycin (20 μM/60 min) then 8-CPT; (4) U0126 (100 nM/20 min) then 8-CPT; (5) PD 98059 (50 μM/90 min), then 8-CPT; (6) LY294002 (20 μM/20 min) then 8-CPT; and (7) rapamycin (100 nM/15 min) then 8-CPT. [3H]Leucine incorporation is reported as the mean ± SE from two experiments performed in triplicate. Panel E: Enumeration of 1-LN prostate cancer cells treated with varying concentration of 8-CPT. Cell number shown are mean ± SE from two experiments performed in duplicate. To the right of Panel E, the bar diagram, Panel F, shows the effect of pretreating the cells with inhibitors of PI 3-kinase or mTOR on 8-CPT-induced increase in cell numbers. The bars are: (1) vehicle; (2) 8-CPT (50 μM); (3) LY294002 (20 μM/20 min), then 8-CPT (50 μM); and (4) rapamycin (100 nM) then 8-CPT. Values significantly different from 8-CPT treated cells at the 5% level are marked with asterisk (*).