Fig. 6.
Effect of AR knockdown on cell proliferation. (A) In vitro proliferation: C4-2 cells were infected with control or AR shRNA lentivirus with enhanced green fluorescent protein marker. Successful knockdown was confirmed by western blot. Cells were incubated in medium with charcoal stripped FBS with BSA or DHA for indicated time. Knockdown of AR significantly slowed down cell proliferation in the medium containing BSA. DHA incubation completely inhibited and knockdown of AR further reduced cell proliferation. Analysis of variance analysis indicates that group a, b, c and d are statistically different (P < 0.05) at day 7. (B) In vivo proliferation: control or AR shRNA infected cells (5 × 105) were mixed with an equal volume of Matrigel, inoculated subcutaneously in pairs into flanks of castrated mice (n = 5) fed with omega-3 PUFA diet. Tumor volumes were monitored once a week and calculated. Mice were terminated at week 6 and tumor weights were determined. Knockdown of AR significantly inhibited tumor growth (P = 0.01, Student's t-test). Bars are standard deviations.