p16重组腺病毒联合顺铂或三氧化二砷对人膀胱癌EJ细胞作用的实验研究

An experimental study of combining Ad-p16 with chemotherapy agents on human bladder cancer cells

目的 探讨 p16重组腺病毒 (Ad p16 )与顺铂 (CDDP)或三氧化二砷 (As2 O3 )联合应用对人膀胱癌EJ细胞体内外生长的抑制效应及机制。 方法 将Ad p16与CDDP或As2 O3 联合应用 ,通过细胞生长抑制实验 ,克隆形成实验 ,细胞周期分析 ,免疫组织化学分析 ,以及裸鼠皮下移植瘤模型 ,观察其对人膀胱癌EJ细胞的作用及机制。 结果 与单独应用相比 ,Ad p16与低剂量的CDDP或As2 O3 联合应用可明显抑制EJ细胞的体外生长 ,诱导EJ细胞凋亡 ,明显阻滞EJ细胞G1期 ;裸鼠体内肿瘤发生时间延迟 ,4周后肿瘤体积与单独应用时相比差别有显著性意义 (P <0 .0 5 )。 结论 Ad p16与CDDP或As2 O3 联合应用 。

Objective To evaluate the therapeutic efficacy of combining p16 expressing adenovirus with chemotherapy agents CDDP or As 2O 3 on human bladder cancer cell line EJ. Methods The human bladder cancer cell line EJ were transfected with adenovirus mediated p16 gene (Ad p16),combining with administration of cisplatin (CDDP) or arsenic trioxide (As 2O 3).The cell growth, morphological changes,cell cycle,apoptosis and molecular changes were measured using cell counting,reversmicroscopy,flow cytometry, cloning formation,immunocytochemical assays and in vivo therapy experiments to evaluate the therapeutic efficacy of such a combined regimen. Results Combined Ad p16 transfer and CDDP or As 2O 3 to EJ cells could exert substantially stronger therapeutic effects than the single chemotherapeutic agent treatment.In vivo experiments,combined administration of p16 and CDDP or As 2O 3 induced much more tumor diminish as compared to the use of the single agent.Massive apoptosis of EJ cell could be obseved by the use of either the combined regimen or the chemotherapeutic agent alone.Cell cycle analysis demonstrated that administration of CDDP or As 2O 3 alone remarkably arrested EJ cell in G2/M prior to apoptotic cell death. When treated with combined regimen, cells were arrested in G1 to a greater extent prior to apoptotic cell death. Conclusions Ad p16 after introduced into EJ cell,shows enhanced therapeutic efficacy on EJ cell when used combinedly with CDDP or As 2O 3.