Han Yali, Miao Jingcheng, Sheng Weihua, Wang Xiaohua, Jing Yingying, Shan Yunbo, Liu Tielian, Bao Wanrong, Yang Jicheng
Cell and Molecular Biology Institute, College of Medicine, Soochow University, Suzhou 215123, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Oct;25(10):1538-45.
To study the inhibitory effect and anti-cancer mechanisms of interleukin 24 (IL-24) on human osteosarcoma cell MG-63, we delivered IL-24 into MG-63 cells in vitro and in vivo by adenovirus. The expression level of IL-24 was detected by RT-PCR and fluorescence microscope; the growth inhibition, apoptosis rate and apoptosis body were measured by MTT, Flow cytometry and Hoechst staining respectively. Furthermore, we analyzed the expression of bcl-2, bax, caspase3 genes by RT-PCR after overexpression of IL-24. For in vivo study, we first established the MG-63 tumor model by grafting MG-63 cells in athymic nude mice; and then injected Ad-IL-24 into the tumors. Two weeks after injection, we sacrificed the mice, removed the tumors, weighed and calculated the ratios of tumor-suppression. We also detected the expressions of Bcl-2, Bax, Caspase-3 and CD34 with immumohistochemistry. Our in vitro results indicated that Ad-IL-24 was transcribed and translated in MG-63 osteosarcoma cells. More interestingly, IL-24 inhibited the growth of MG-63 cells and induced apoptosis by up-regulation of bax, caspase-3 and down-regulation of bcl-2. The in vivo data showed that IL-24 suppressed the tumor growth conspicuously through down-regulating the expression of bcl-2, and up-regulating the expression of bax, caspase-3. This study would provide evidence for the gene therapy of IL-24 on osteosarcoma.
为研究白细胞介素24(IL-24)对人骨肉瘤细胞MG-63的抑制作用及抗癌机制,我们通过腺病毒在体外和体内将IL-24导入MG-63细胞。采用RT-PCR和荧光显微镜检测IL-24的表达水平;分别用MTT、流式细胞术和Hoechst染色检测细胞生长抑制、凋亡率和凋亡小体。此外,在IL-24过表达后,通过RT-PCR分析bcl-2、bax、caspase3基因的表达。对于体内研究,我们首先通过将MG-63细胞接种到无胸腺裸鼠体内建立MG-63肿瘤模型;然后将Ad-IL-24注射到肿瘤中。注射两周后,处死小鼠,取出肿瘤,称重并计算抑瘤率。我们还用免疫组织化学检测了Bcl-2、Bax、Caspase-3和CD34的表达。我们的体外实验结果表明,Ad-IL-24在MG-63骨肉瘤细胞中进行转录和翻译。更有趣的是,IL-24通过上调bax、caspase-3和下调bcl-2抑制MG-63细胞的生长并诱导凋亡。体内实验数据表明,IL-24通过下调bcl-2的表达和上调bax、caspase-3的表达显著抑制肿瘤生长。本研究将为IL-24对骨肉瘤的基因治疗提供依据。