Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei Province, People's Republic of China.
Eur J Pharmacol. 2010 Dec 15;649(1-3):120-6. doi: 10.1016/j.ejphar.2010.09.040. Epub 2010 Sep 24.
This study was purposed to explore the mechanism of augmenting apoptosis-inducing effects of cisplatin on colorectal cancer SW480 cells by stable transfection of Programmed Cell Death 5 (PDCD5) gene, both in vitro and in vivo. The expression level of PDCD5 of SW480 cells stably transfected with PDCD5 (SW480/PDCD5) and pcDNA3.1/Neo(+) empty vector stably transfected cells (SW480/Neo) was examined by real-Time RT-PCR and Western blot. The effects of cisplatin in combination with PDCD5 on the proliferation and apoptosis of colorectal cancer cells were measured by using MTT analysis, Hoechst 33258 analysis and flow cytometry with Annexin-V-FITC/PI dual labeling, respectively. To examine the combination therapeutic effect of PDCD5 and cisplatin, tumor xenograft model was prepared for in vivo study. The results showed that PDCD5 levels of SW480/PDCD5 were higher than SW480 and SW480/Neo, respectively. The PDCD5 enhanced the apoptotic percentage of SW480/PDCD5 cells induced by cisplatin, as compared SW480 and SW480/Neo, respectively. In in vivo study, the transfection of PDCD5 increased the efficacy of cisplatin-induced inhibition of tumor growth in nude mice. It is concluded that stable transfection of PDCD5 gene can notably improve apoptosis-inducing effects of cisplatin on colorectal cancer cells, which is a novel strategy to better chemotherapeutic effects on colorectal cancer.
本研究旨在通过稳定转染程序性细胞死亡因子 5(PDCD5)基因,探讨增强顺铂诱导结直肠癌细胞 SW480 细胞凋亡作用的机制,包括在体外和体内。通过实时 RT-PCR 和 Western blot 检测稳定转染 PDCD5(SW480/PDCD5)和 pcDNA3.1/Neo(+)空载体稳定转染细胞(SW480/Neo)的 PDCD5 表达水平。采用 MTT 分析、Hoechst 33258 分析和 Annexin-V-FITC/PI 双标记流式细胞术分别检测顺铂与 PDCD5 联合对结直肠癌细胞增殖和凋亡的影响。为了检测 PDCD5 与顺铂联合治疗的效果,制备了肿瘤异种移植模型进行体内研究。结果显示,SW480/PDCD5 的 PDCD5 水平高于 SW480 和 SW480/Neo,PDCD5 增强了顺铂诱导的 SW480/PDCD5 细胞的凋亡百分比,与 SW480 和 SW480/Neo 相比。在体内研究中,转染 PDCD5 增加了顺铂抑制裸鼠肿瘤生长的疗效。结论:稳定转染 PDCD5 基因可显著提高顺铂诱导结直肠癌细胞凋亡的作用,为改善结直肠癌的化疗效果提供了一种新策略。