Chen Jie, Hu Jun, Dong Chang-yuan, Liang Ke, Dai Ying, Gao Jing
State Key Laboratory of Virology, Wuhan University School of Medicine, China.
Zhonghua Zhong Liu Za Zhi. 2007 Jul;29(7):505-9.
To study the death mode of human hepatocellular carcinoma Hep-3B cells and human lung adenocarcinoma A549 cells induced by bluetongue virus strain HbC3 (BTV-HbC3) and the mechanism of its action.
BTV-HbC3 was used to infect the tumor cells, and the cytopathic effects (CPE) was observed. TUNEL staining was used to detect the apoptosis of tumor cells induced by BTV-HbC3. The changes of endoplasmic reticulum and nuclei treated with BTV-HbC3 were further examined by laser scanning confocal microscopy. The activities of caspase-3/7, caspase-8 and caspase-9 were determined by fluorescence analysis.
Hep-3B cells were sensitive to BTV-HbC3. Lots of early apoptotic cells were found by TUNEL staining. The laser scanning confocal microscopic examination showed characteristics of apoptosis, such as pyknotic nuclei, margination of nuclear chromatin and vacuolization of endoplasmic reticulumin in Hep-3B cells exposed to BTV-HbC3. The activity of caspase-3/7 was increased, but the activity changes of caspase-8 and caspase-9 were not found. A549 cells were sensitive to BTV-HbC3 too. But no apoptotic cells were observed by TUNEL staining. The results of laser scanning confocal microscopy showed marked vacuolization of endoplasmic reticulum, but chromatin margination was not found after A549 cells was exposed to BTV-HbC3. The activity of caspase-3/7 and caspase-9 was increased, but the activity of caspase-8 was not changed.
BTV-HbC3 induces apoptosis of Hep-3B tumor cells mainly through endoplasmic reticulum signal transduction pathway, and the features of cell death in A549 cells could be described as paraptosis.
研究蓝舌病毒HbC3株（BTV-HbC3）诱导人肝癌Hep-3B细胞和人肺腺癌A549细胞的死亡方式及其作用机制。
用BTV-HbC3感染肿瘤细胞，观察细胞病变效应（CPE）。采用TUNEL染色检测BTV-HbC3诱导的肿瘤细胞凋亡。通过激光扫描共聚焦显微镜进一步检测经BTV-HbC3处理后的内质网和细胞核的变化。采用荧光分析法测定caspase-3/7、caspase-8和caspase-9的活性。
Hep-3B细胞对BTV-HbC3敏感。TUNEL染色发现大量早期凋亡细胞。激光扫描共聚焦显微镜检查显示，暴露于BTV-HbC3的Hep-3B细胞具有凋亡特征，如核固缩、核染色质边缘化和内质网空泡化。caspase-3/7活性增加，但未发现caspase-8和caspase-9活性变化。A549细胞对BTV-HbC3也敏感。但TUNEL染色未观察到凋亡细胞。激光扫描共聚焦显微镜结果显示，A549细胞暴露于BTV-HbC3后内质网出现明显空泡化，但未发现染色质边缘化。caspase-3/7和caspase-9活性增加，但caspase-8活性未改变。
BTV-HbC3主要通过内质网信号转导途径诱导Hep-3B肿瘤细胞凋亡，A549细胞的死亡特征可描述为胀亡。
