鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶双反义腺病毒对大肠癌细胞生长的抑制作用

Inhibitory effect of ornithine decarboxylase and S-adenosylmethionine decarboxylase biantisense adenovirus on colorectal cancer cells

目的:研究鸟氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸脱羧酶(AdoMetDC)双反义腺病毒(Ad-ODC-AdoMetDCas)对大肠癌细胞生长的抑制作用,并初步探讨其参与细胞周期调节的分子机制。方法:将Ad-ODC-AdoMetDCas感染大肠癌细胞株HT-29,通过MTS法观察其对HT-29细胞增殖的影响,流式细胞术检测其对细胞周期的影响,采用Western blot检测Ad-ODC-AdoMetDCas对细胞中ODC、AdoMetDC及细胞周期调节蛋白(Cyclin D1 and CDK4)的表达的影响,利用RT-PCR检测Ad-ODC-AdoMetDCas对细胞周期蛋白Cyclin D1的mRNA水平的影响。结果:Ad-ODC-AdoMetDCas可抑制HT-29细胞中ODC和AdoMetDC蛋白的表达,并可明显抑制细胞的增殖,引起细胞周期Gl期阻滞,抑制G1期主要的细胞周期蛋白Cyclin D1的转录和翻译。结论:Ad-ODC-AdoMetDCas可有效下调ODC和AdoMetDC的表达,并通过抑制Cyclin D1的表达使其细胞周期停滞于G1期,从而抑制大肠癌细胞HT-29的增殖,为进一步研究大肠癌的基因治疗打下基础。

Objectiue： To investigate the inhibitory effect of Ad-ODC-AdoMetDCas which can simultaneously express both antisense ornithine decarboxylase （ODC） and S-adenosylmethionine decarboxylase （AdoMetDC） on colorectal cancer cell growth and detect the possible mechanism, Methede： Human colorectal cancer cells HT-29 were cultured in RPMI 1640 medium and were infected with Ad-ODC-AdoMetDCas, MTS was used to assess the HT-29 cell growth. Cell cycle progression was detected by flow cytometry analysis, ODC, AdoMetDC and cell cycle regulated proteins levels were measured by Western blot analysis, The mRNA level of cyclin D1 was measured by RT-PCR. Results： The recombinant adenovirus Ad-ODC-AdoMetDCas downregulated the expression of ODC and AdoMetDC in HT-29 cells. Ad-ODC-AdoMetDCas significantly inhibited the cell growth and induced G1 arrest. Cyclin DI protein and mRNA levels were decreased by Ad-ODC-AdoMetDCas infection, Conclusion： Downregulation of ODC and AdoMetDC mediated by Ad-ODC-AdoMetDCas induces G1 arrest in HT-29 cells by suppressing cyclin DI expression. As a new anticancer reagent, Ad-ODC-AdoMetDCas has potential therapeutic effect on colorectal cancers.