Chi Weiling, Song Xianrang, Jiang Chunying, Liu Xianxi, Li Wentong, Wang Xingwu
Institute of Biochemistry and Molecular Biology, Shandong University School of Medicine, Jinan, China.
Tumour Biol. 2006;27(5):243-51. doi: 10.1159/000094843. Epub 2006 Jul 28.
Ornithine decarboxylase (ODC), a molecule that is overexpressed in various cancers, has been associated with cell growth and proliferation. Downregulation of ODC may inhibit tumor cell growth. To verify this hypothesis, ODC gene silencing was studied in colorectal cancer cells in vitro and in vivo. A lentiviral system harboring both enhanced green fluorescent protein reporter gene and the ODC short hairpin RNA expression cassette was firstly constructed. Then the stable ODC gene silencing cell lines HT29/ODC and LoVo/ODC were established and their growth-inhibiting characteristics were identified. Real-time RT-PCR analysis of mRNA and Western blot analysis of proteins were used to evaluate the expression of the ODC gene. In the cells studied, ODC expression was significantly decreased. Proliferation of HT29/ODC and LoVo/ODC cells was obviously retarded. In animal experiments, HT29/ODC and LoVo/ODC cell xenografts demonstrated growth suppression compared to parental or control colorectal cancer cell xenografts. The results demonstrated that lentiviral vector was capable of downregulating ODC, resulting in impressive anticancer effects. It offers a powerful new strategy for cancer gene therapy and may be useful in the control of solid tumors, such as human colorectal carcinomas.
鸟氨酸脱羧酶（ODC）是一种在多种癌症中过度表达的分子，与细胞生长和增殖有关。ODC的下调可能会抑制肿瘤细胞的生长。为了验证这一假设，我们在体外和体内的结肠癌细胞中研究了ODC基因沉默。首先构建了一个携带增强型绿色荧光蛋白报告基因和ODC短发夹RNA表达盒的慢病毒系统。然后建立了稳定的ODC基因沉默细胞系HT29/ODC和LoVo/ODC，并鉴定了它们的生长抑制特性。使用实时RT-PCR分析mRNA和蛋白质的Western印迹分析来评估ODC基因的表达。在所研究的细胞中，ODC表达显著降低。HT29/ODC和LoVo/ODC细胞的增殖明显受到抑制。在动物实验中，与亲本或对照结肠癌细胞异种移植相比，HT29/ODC和LoVo/ODC细胞异种移植表现出生长抑制。结果表明，慢病毒载体能够下调ODC，产生显著的抗癌效果。它为癌症基因治疗提供了一种强大的新策略，可能对控制实体瘤如人类结肠癌有用。
