Ad-IL-24对人胶质瘤细胞生长抑制效应的体外实验

Adenovirus mediated IL-24 gene expression inhibits growth of human glioma cell in vitro

研究携带人白介素24(IL-24)的腺病毒表达载体(Ad-IL-24)对人U251胶质瘤细胞生长的影响和抗肿瘤分子机制。将不同MOI Ad-IL-24感染U251人胶质瘤细胞后,MTT法检测Ad-IL-24对U251细胞生长的抑制作用,流式细胞仪和Hochest染色法检测细胞的凋亡率。RT-PCR检测bcl-2、bax、ICE、C-myc、HIF-1α和p53等基因的转录表达水平,Western blotting检测Cleaved Caspase-3的表达。结果表明100 MOI Ad-IL-24感染U251细胞后能明显抑制细胞生长,并能明显诱导细胞凋亡,感染72 h后细胞凋亡率可达42%,感染4 d后细胞生长抑制率可达50%。RT-PCR检测发现Ad-IL-24能引起与细胞凋亡和血管形成相关基因bax/bcl-2、ICE、C-myc、p53的上调和HIF-1α的下调,并促进Caspase-3的活化。本研究结果显示Ad-IL-24能明显抑制人胶质瘤细胞U251生长和诱导细胞凋亡,其抗肿瘤机制可能与通过bax/bcl-2、ICE、c-myc、p53的上调和HIF-1α的下调,进而导致Caspase-3的活化而诱导肿瘤细胞发生凋亡有关。

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-la, 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.