Objective To investigate whether Topoisomerase (DNA) II alpha (TOP2α) promote prostate cancer cells proliferation and regulated by p53.Methods After inhibiting expression of p53 by interfering RNA (RNAi) in C4-2 cells using Lipofectamine2000,the mRNA and protein expression level were assessed by Real-time PCR and Western Blot.After transfecting GFP-p53 in 293T cells by according to the manufacturer''s instruction,the expression level of TOP2α was assessed by Western Blot.After knockdown TOP2α by RNAi in C4-2 cells,proliferation was tested by Bromodeoxyuridine incorporation assay (BrdU).Then,the BrdU assay was repeated after transfecting GFP-TOP2α.Results The mRNA and protein level were significant increased after transfection of siRNA against p53.Moreover,proliferation was further decreased from (21.54±1.443)% to(13.78±0.4736)% (P<0.05).At the meantime,after transfectiong GFP-TOP2α,proliferation was further increased from (20.84±2.082%)to(33.36±3.244)% (P<0.05).Conclusion Our findings suggest that p53 regulates the mRNA and protein level of TOP2α,and TOP2α could regulates proliferation ability of C4-2 cells.%目的 观察TOP2α对前列腺癌细胞增殖能力的影响以及抑癌基因p53对TOP2α的调控能力.方法 以Lipofectamine2000试剂转染针对p53和非特异性对照组的siRNA,干扰前列腺癌细胞C4-2中p53 的表达,采用Real-time PCR和Western Blot检测细胞内TOP2α的mRNA和蛋白水平表达变化;采用质粒转染法在293T细胞中过表达p53后检测TOP2α蛋白水平的变化,明确p53对TOP2α表达的影响;采用BrdU标记法计算并检测干扰或过表达TOP2α表达后对前列腺癌细胞增殖能力的影响,明确TOP2α对前列腺癌细胞增殖的调节能力.结果 干扰p53表达后细胞内TOP2α mRNA的表达明显升高,说明p53可以调节TOP2α的转录水平,进一步检测干扰p53表达后,TOP2α的蛋白水平表达明显升高;在细胞内,过表达p53后检测TOP2α的蛋白表达水平明显下降;在C4-2细胞中干扰目标基因TOP2α的表达后,增殖细胞的百分比从(21.54±1.443)%下降至(13.78±0.4736)%(P<0.05).同时,在C4-2细胞中过表达TOP2α后,增殖细胞的百分比从(20.84±2.082%)上升至(33.36±3.244)% (P<0.05).结论 在前列腺癌细胞中,p53可以在mRNA和蛋白水平调节TOP2α的表达,TOP2α具有调节前列腺癌细胞增殖的能力.
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