Ad-IL-24对SGC-7901胃癌细胞生长抑制的体外实验

Adenovirus mediated IL-24 gene expression suppresses gastric cancer cell growth in vitro

摘要:

旨在研究携带人IL-24基因的腺病毒表达载体(Ad-IL-24)对SGC-7901人胃癌细胞的生长抑制作用并分析其分子机制。以不同MOI(感染复数)的Ad空载体腺病毒感染SGC-7901人胃癌细胞, 筛选出最佳感染剂量; Ad-IL-24以最佳感染剂量感染SGC-7901胃癌细胞, RT-PCR法和Western blotting法检测腺病毒介导的IL-24基因在SGC-7901胃癌细胞中的转录; MTT法检测Ad-IL-24对SGC-7901胃癌细胞的生长抑制作用, 流式细胞仪检测其诱导SGC-7901人胃癌细胞凋亡和细胞周期改变的效应, Hoechst33258染色荧光显微镜检测其诱导胃癌细胞凋亡的核形态改变; RT-PCR半定量法进一步检测SGC-7901胃癌细胞中凋亡相关基因的转录。结果显示, 100 MOI 为感染SGC-7901胃癌细胞的腺病毒最佳感染剂量; Ad-IL-24能成功介导IL-24基因在SGC-7901胃癌细胞中转录性表达; Ad-IL-24感染SGC-7901胃癌细胞后, 能明显抑制胃癌细胞生长和诱导凋亡; Ad-IL-24能显著上调SGC-7901胃癌细胞中bax、caspase-3和p53的表达和下调bcl-2的表达。因此, 腺病毒介导的IL-24表达具有明显抑制SGC-7901人胃癌细胞生长和诱导凋亡的抗肿瘤效应, 其机制可能与上调bax/bcl-2、caspase-3和p53密切相关。

Abstract:

To study the inhibitory effect of a recombinant adenoviral vector carrying human IL-24 gene on SGC-7901 human gastric cancer cell. We infected the SGC-7901 gastric cancer cells with Ad blank adenovirus at various multiplicity of infection (MOIs) to find the optimal infective dose. The SGC-7901 tumor cells were infected with Ad-IL-24 at the optimal MOI in the following experiments. Adenovirus-mediated IL-24 transcription expression in SGC-7901cells was examined by RT-PCR. The growth-suppressing effect of Ad-IL-24 on SGC-7901 tumor cells was assessed by MTT assay. Apoptosis and cell cycle of SGC-7901 tumor cells infected with Ad-IL-24 was evaluated by flow cytometer (FCM), respectively. The karyomorphology of apoptotic SGC-7901 tumor cells was examined using Hoechst33258 staining under fluorescence microscopy. The expression of apoptosis-related genes was future determined by semi-quantification RT-PCR; We demonstrated that the MOI of 100 was the optimal infective dose in the study on adenovirus-mediated IL-24 gene transfer into SGC-7901 gastric cancer cell; IL-24 gene mediated by adenovirus could successfully transcribe in SGC-7901 tumor cells; Ad-IL-24 could significantly inhibit SGC-7901 tumor cell growth and induce apoptosis, it also can up-regulate the express of bax, caspase-3 and p53 whilst down-regulate the bcl-2 expression. Thus, adenovirus-mediated IL-24 expression had marked anti-tumor effect in suppressing SGC-7901 human gastric cancer cell growth and inducing apoptosis, which may be closely associated with its up-regulation of bax/bcl-2, caspase-3 and p53.