目的观察TAT-N24穿膜融合多肽抑制前列癌细胞增殖的作用。方法用纯化的TAT-N24穿膜融合多肽处理前列癌PC3细胞,采用流式细胞仪检测细胞周期进程的改变,BrdU掺入法检测其对细胞DNA合成的影响,Western blot法检测细胞内AKT蛋白的磷酸水平的变化。结果①细胞周期分布:空白组G0/G1期(56.9±6.1)%,S期(27.0±2.3)%,G2/M期(16.1±1.3)%;对照多肽组G0/G1期(57.9±4.8)%,S期(24.2±3.1)%,G2/M期(27.8±1.4)%;TAT-N24组G0/G1期(68.6±5.4)%(P〈0.05),S期(19.4±1.5)%(P〈0.05),G2/M期(12.3±1.4)%。②BrdU阳性细胞为空白组(37.9±3.2)%,对照多肽组(36.2±4.1)%,TAT-N24组(21.5±2.4)%(P〈0.05);③AKT磷酸化水平组间差异无统计学意义。结论 TAT-N24穿膜融合多肽能有效阻滞前列腺癌PC3细胞的细胞周期进程,并抑制其DNA合成。TAT-N24穿膜融合多肽有望成为有效的治疗前列腺癌的分子靶向药物。
Objective To investigate the anti-proliferation effects of cell-permeable TAT-N24 fusion peptide on prostate cancer cells. Methods The human prostate cancer cell line PC3 was treated with TAT-N24,cell cycle distribution was analyzed by flow cytometry,DNA synthesis of which was measured by BrdU incorporation assay.and the expression of phosphorylated AKT was examined by Western blot analysis. Results ① G0/G1 phase arrest induced by TAT-N24 was observed.The cell cycle distribution was that: in the blank group G0/G1 phase as(56.9±6.1)%,S phase(27.0±2.3)%,G2/M phase(16.1±1.3)%;G0/G1 phase(57.9±4.8)%,S phase(24.2±3.1)%,G2/M phase(27.8±1.4)% in the control peptide group;G0/G1 phase(68.6±5.4)%(P0.05),S phase(19.4±1.5)%(P0.05),G2/M phase(12.3±1.4)% in TAT-N24 group,respectively.②The BrdU positive cells in the blank,control peptide and TAT-N24 groups was(37.9±3.2)%,(36.2±4.1)%,(21.5±2.4)%(P0.05),respectively;③It was revealed that TAT-N24 had no significant effects on AKT phosphorylation Conclusion The cell-permeable TAT-N24 fusion peptide effectively blocks cells processing into G0/G1 phase and inhibiting DNA synthesis of PC3 cell line.TAT-N24 may become an effective molecular targeted gene medicine for cancer therapy.