PCNA反义寡核苷酸抑制人膀胱癌EJ细胞增殖活性的实验研究  

The effect of proliferating cell nuclear antigen-specific antisense oligonucleotide on human bladder cancer cell line

作　　者：曾甫清[1] 朱朝晖[1] 林晨 赵军[1] 鲁功成[1] 

机构地区：[1]同济医科大学附属协和医院泌尿外科,武汉430022 [2]中国医学科学院肿瘤研究所分子肿瘤国家重点实验室

出　　处：《中华泌尿外科杂志》2000年第4期213-215,共3页Chinese Journal of Urology

基　　金：卫生部科研基金!( 96 2 111)

摘　　要：目的　探讨全硫代修饰型PCNA反义寡核苷酸 (ASO)对人膀胱癌EJ细胞体外增殖活性及裸鼠体内致瘤性的影响。　方法　PCNA ASO在脂质体介导下转染EJ细胞后 ,运用细胞计数、MTT法、3 H TdR掺入试验、流式细胞术 (FCM)、克隆形成率 (CF)、SABC免疫组化、建立裸鼠动物模型等方法分析其体内外抗瘤活性及机制。　结果　在脂质体介导下 ,0 4～ 2 0 μmol/LPCNA ASO对人膀胱癌EJ细胞体外生长有一定程度的抑制作用 (31 4%～ 78 6 % ) ,呈现时间及计量效应 ;DNA合成速率降低 45 7%～ 84 6 % ;克隆形成能力下降 5 4 3%～ 78 8% ;PCNA蛋白表达减少 ;S期比率显著低于对照组 ,细胞生长受阻于G0 /G1期。　结论　PCNA ASO可抑制EJ细胞体外生长 ,降低DNA合成速率 ,减少PCNA蛋白表达 ;在裸鼠体内 ,PCNA ASO具有抗瘤活性。Objective To assess the effects of antisense oligonucleotide specific for proliferating cell nuclear antigen(PCNA)mRNA on human bladder cancer cell line EJ. Methods 18 mer phosphorothioate PCNA specific oligonucleotide was encapsulated by lipofectin and then added into culture media of human bladder cancer cell EJ.The cell growth,cell cycle analysis,colony formation and PCNA protein production were observed. Results When treated with PCNA antisense oligonucleotide at concentrations of 0.4～ 2.0 μmol/L with dose and time dependently,the growth and proliferation of EJ cells were inhibited.Whereas, cycle analysis showed a significant decrease in S phase and a remarkable increase in G 1/G 0 fraction after exposure to PCNA ASO as comparison with the cell treated with sense or scramble oligonucleotide at the corresponding time. Conclusions The results indicate that PCNA is essential for human bladder cancer cell proliferation. PCNA ASO encapsulated by lipofectin exerts a strong inhibitory effect on human bladder cancer EJ cell line.Therefore,PCNA specific antisense oligonucleotide may influence the behavior of human bladder cancer cell EJ.

关 键 词：膀胱肿瘤 增殖细胞核抗原 反义寡核苷酸 

分 类 号：R737.14[医药卫生—肿瘤]