Shan Yunbo, Sheng Weihua, Xie Yufeng, Liu Tielian, Jing Yingying, Hu Zhiqing, Yang Jicheng
Department of Celluar and Molecular Biology, Medical School, Suzhou University, Suzhou 215123, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Feb;25(2):279-86.
To investigate the inhibitory effect and anti-cancer mechanism of adenovirus mediated IL-24 gene expression on the human U251 glioma cell. U251 glioma cells were infected with Ad-IL-24 at various multiplicity of infection (MOIs). Cell proliferation was determined by MTT assay. Cell apoptosis was detected by flow cytometry and Hochest staining. The transcription of apoptosis-related genes was analyzed by reverse transcription-PCR (RT-PCR), and the expression of Cleaved Caspase-3 was analyzed by Western blotting. The result showed that the growth of U251 glioma cells was significantly inhibited by Ad-IL-24 at the MOI of 100. The apoptotic rate of U251 glioma cells was 42% 72 h after infection with Ad-IL-24. Four days after infection, the growth of the U251 glioma cells was inhibited to 50%. RT-PCR showed that Ad-IL-24 not only up-regulated expression of bax/bcl-2, ICE, C-myc, p53 and down-regulated the expression of HIF-1alpha, but also enhanced Caspase-3 activation, eventually resulting apoptosis. Taken together, these results suggest that infection of U251 glioma cells with Ad-IL-24 can inhibit growth and induce apoptosis significantly by the regulation of apoptosis-related genes.
探讨腺病毒介导的IL-24基因表达对人U251胶质瘤细胞的抑制作用及抗癌机制。用不同感染复数（MOIs）的Ad-IL-24感染U251胶质瘤细胞。采用MTT法检测细胞增殖。通过流式细胞术和Hochest染色检测细胞凋亡。用逆转录-聚合酶链反应（RT-PCR）分析凋亡相关基因的转录情况，并用蛋白质免疫印迹法分析Cleaved Caspase-3的表达。结果显示，在感染复数为100时，Ad-IL-24可显著抑制U251胶质瘤细胞的生长。感染Ad-IL-24 72小时后，U251胶质瘤细胞的凋亡率为42%。感染四天后，U251胶质瘤细胞的生长被抑制至50%。RT-PCR结果显示，Ad-IL-24不仅上调了bax/bcl-2、ICE、C-myc、p53的表达，下调了HIF-1α的表达，还增强了Caspase-3的激活，最终导致细胞凋亡。综上所述，这些结果表明，Ad-IL-24感染U251胶质瘤细胞可通过调节凋亡相关基因显著抑制其生长并诱导凋亡。
