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重组腺病毒介导KDRP-CD/TK基因对大肠癌细胞增殖与凋亡的影响[J]. 肿瘤防治研究, 2008, 35(03): 173-176. DOI: 10.3971/j.issn.1000-8578.1839
引用本文: 重组腺病毒介导KDRP-CD/TK基因对大肠癌细胞增殖与凋亡的影响[J]. 肿瘤防治研究, 2008, 35(03): 173-176. DOI: 10.3971/j.issn.1000-8578.1839
Effect of Adenovirus Mediated Fusion Gene System Driven by KDR Promoter on Proliferation and Apoptosis of Colorectal Cancer Cells[J]. Cancer Research on Prevention and Treatment, 2008, 35(03): 173-176. DOI: 10.3971/j.issn.1000-8578.1839
Citation: Effect of Adenovirus Mediated Fusion Gene System Driven by KDR Promoter on Proliferation and Apoptosis of Colorectal Cancer Cells[J]. Cancer Research on Prevention and Treatment, 2008, 35(03): 173-176. DOI: 10.3971/j.issn.1000-8578.1839

重组腺病毒介导KDRP-CD/TK基因对大肠癌细胞增殖与凋亡的影响

Effect of Adenovirus Mediated Fusion Gene System Driven by KDR Promoter on Proliferation and Apoptosis of Colorectal Cancer Cells

  • 摘要: 目的探讨腺病毒介导KDR启动子驱动的融合基因体系对人大肠癌细胞LOVO的增殖活性及凋亡的影响。方法以重组腺病毒AdEasy-KDR-CDglyTK体外感染表达KDR的LOVO细胞株和对照组不表达KDR的LS174T细胞株,观察该体系对LOVO细胞的杀伤效应;并以流式细胞仪检测细胞周期的变化,电镜观察细胞的病变。结果重组腺病毒对LOVO细胞及LS174T细胞的感染率相似,其感染率随腺病毒滴度的递增而增加,当MOI为200时,所有细胞株均达约100%感染。以MOI为100的重组体分别感染各细胞株表现出对前药的不同敏感性:表达KDR的LOVO细胞对前药的具有较高的敏感性,与前者相比,不表达KDR的LS174T细胞对前药不敏感(P均〈0.001)。融合基因的疗效优于任一单自杀基因(P均〈0.001)。将感染腺病毒的细胞与未感染细胞以不同混合培养,观察到该体系明显的旁观者效应。流式细胞术检测表明该体系抑制LOVO细胞DNA的合成,表现为S期阻止。同时,电镜下可见LOVO有凋亡和坏死改变。结论KDR基因启动子可调控融合基因体系选择性杀伤人大肠癌LOVO细胞,其机制与细胞周期阻滞、凋亡及坏死有关。

     

    Abstract: Objective  To study the effect of adenovirus (Ad) mediated double suicide gene driven by KDR promoter on proliferation and apoptosis of human colorectal cancer LOVO cells. Methods  The KDR producing cells LOVO and the KDR nonproducing cells LS174 T were infected by the AdEasy2KDR2CDg2 lyTK respectively, followed by t reatment with 52FC and GCV. killing effect s of this suicide gene system on colorectal cancer cells were evaluated. Moreover, the dist ribution of cell cycle was detected by flow cytomet ric assay and pathological character of cells was observed by elect ron microscopy. Results  The infection rate of the resultant recombinant Ad ( rAd) to all the cells had not apparently different, and it in2 creased gradually with the addition of multiple of infection (MOI) of Ads. The infected cells exhibited dif2 ferent sensibility to the two prodrugs : LOVO cells infected with rAd were sensitive highly to the pro2 drugs, but the LS174 T cells infected with rAd appeared to be unsensitive to the two prodrugs ( P <0. 001) . The killing effect of CD/ TKfusion gene on the target cells was much st ranger than that of each single suicide gene ( P < 0. 001), and they all had considerable bystander effect . In addition, the cell cycle of LOVO cells was arrested at S phase and morphologic features of apoptosis and necrosis in LOVO cells were displayed via elect ron microscopy. Conclusion  The CD/ TK fusion gene system cont rolled by KDR promoter has selectively killing effect on the KDR2CDglyTKexpressing LOVO cells, the mechanism of which may involve the halting the cell cycles and cytotoxic necrosis and apoptosis of cell.

     

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