p53重组腺病毒联合增殖细胞核抗原反义寡核苷酸治疗膀胱癌的研究

Proliferating cell nuclear antigen antisense oligonucleotide in combined with recombinant adenovirus p53 to treat bladder cancer

目的 探讨重组腺病毒p53(Ad p53)与增殖细胞核抗原反义寡核苷酸 (PCNA ASO)联合应用对膀胱癌的抑瘤效应。方法 用腺病毒将p53 [感染强度 (MOI) 1 0 0 ]导入细胞 ,PCNA ASO(1 .6μmol/L)在脂质体 (Lipofectin)介导下转染细胞 ,通过噻唑蓝比色法 (MTT)、流式细胞术、克隆形成、建立移植瘤模型 ,探讨其体内外抗瘤活性。结果 Ad p53与PCNA ASO联合应用明显抑制膀胱癌胞EJ(89.3 % )和BIU 87(78.6 % )生长 ,细胞克隆形成能力分别下降 74.8%和 67.5 % ,S期比率 (EJ细胞 1 1 .4% ,BIU 87细胞 1 4 .6 % )明显降低 ,细胞生长受阻于G1 期 (EJ细胞 62 .2 % ,BIU 87细胞 56 .8% ) ;联合应用 7d后 ,肿瘤体积较初始分别下降 47.6 %和 36 .4%。结论 Ad p53联合PCNA ASO可抑制膀胱癌细胞体内外生长 。

Objective To evaluate the antitumor efficacy of proliferating cell nuclear antigen antisense oligonucleotide (PCNA ASO) in combination with recombinant adenovirus p53 against bladder cancer EJ and BIU 87 cells in vitro and in vivo.Methods Cells were transfected with Ad p53 (100 MOI). Then,PCNA ASO (1.6 μmol/L) with a cationic lipid (lipofectin,20 ml/L) was added to the cells. In vitro and in vivo antitumor effect of PCNA ASO in combination with Ad p53 were measured using MTT assay,flow cytometry,cloning formation and in nude mice model.Results The combination of PCNA ASO and Ad p53 inhibited the cell viability in EJ (89.3%) and BIU 87 (78.6%). The abilities to form foci were decreased by 74.8% in EJ and 67.5% in BIU 87 respectively. A significant decrease was showed at S phase (11.4% in EJ,14.6% in BIU 87) and a remarkable increase at G 1 phase (62.2% in EJ,56.8% in BIU 87). The mean tumor volume after treatment for 7 days with PCNA ASO in combination with Ad p53 was decreased to 47.6% or 36.4% of the initial tumor size,respectively.Conclusion Combined use of PCNA ASO and Ad p53 could inhibit the growth of bladder cancer in vitro and nude mice transplant tumor in vivo,producing synergic effect.