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目的:利用RNA干扰(RNA i)技术,阻断前列腺癌LNCap细胞中类固醇受体共激活分子(SRC-1)的表达,并研究SRC-1基因沉默后对LNCap细胞的影响。方法:将设计好的siRNA,用脂质体法转染LNCap细胞,通过Q-PCR、蛋白免疫印迹检测SRC-1的表达变化,并用CCK-8法检测细胞生长情况的变化。结果:转染SRC-1siRNA 24 h后的前列腺癌LNCap细胞中SRC-1 mRNA的量与空白对照组相比较下降了约35%,转染48 h后下降了约77%,差异具有统计学意义(P<0.05)。转染24~48 h后LNCap细胞中SRC-1蛋白的表达量(24 h为0.359±0.034;48 h为0.257±0.065)与阴性对照组(24 h为0.782±0.078;48 h为0.766±0.043)、转染试剂组(24 h为0.840±0.013;48 h为0.786±0.051)和空白对照组(24 h为0.816±0.065;48 h为0.805±0.107)相比较明显减少(P<0.05)。转染24、48、72、96 h后LNCap细胞的生长抑制率分别为25%、52%、55%和60%。结论:SRC-1与LNCap细胞的生长相关。SRC-1在激素非依赖性前列腺癌中的高表达可能参与了前列腺癌雄激素非依赖性的转变过程。抑制SRC-1的表达可能成为临床治疗激素依赖性前列腺癌的方法之一。
OBJECTIVE: To block the expression of steroid receptor co-activator (SRC-1) in prostate cancer LNCap cells by RNA interference (RNAi) technology and to study the effect of SRC-1 gene silencing on LNCap cells. Methods: The designed siRNA was transfected into LNCap cells by lipofectamine. The expression of SRC-1 was detected by Q-PCR and Western blot. The cell growth was detected by CCK-8. Results: The expression of SRC-1 mRNA in prostate cancer LNCap cells transfected with SRC-1 siRNA for 24 h decreased by 35% compared with the control group, and decreased by 77% after 48 h of transfection, the difference was statistically significant (P <0.05). The expression of SRC-1 protein in LNCap cells 24 hours after transfection (0.359 ± 0.034 at 24 h and 0.257 ± 0.065 at 48 h) was significantly lower than that in the negative control group at 24 h (0.782 ± 0.078 at 24 h and 0.766 ± 0.043 at 48 h ) In transfection reagent group (0.840 ± 0.013 at 24 h and 0.786 ± 0.051 at 48 h), and significantly decreased compared with blank control group (0.816 ± 0.065 at 24 h and 0.805 ± 0.107 at 48 h) (P <0.05). The growth inhibition rates of LNCap cells were 25%, 52%, 55% and 60% respectively at 24, 48, 72 and 96 hours after transfection. Conclusion: SRC-1 is associated with the growth of LNCap cells. The high expression of SRC-1 in hormone-independent prostate cancer may be involved in the androgen-independent transition of prostate cancer. Inhibiting the expression of SRC-1 may become one of the methods for clinical treatment of hormone-dependent prostate cancer.