Involvement of caspase 8 in apoptosis induced by ultrasound-activated hematoporphyrin in sarcoma 180 cells in vitro
Involvement of caspase 8 in apoptosis induced by ultrasound-activated hematoporphyrin in sarcoma 180 cells in vitro
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DOI：
10.7863/jum.2008.27.4.645
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发表时间：
2008-04-01
影响因子：
2.3
通讯作者：
Zhang, Jing
中科院分区：
文献类型：
--
作者：
Tang, Wei;Liu, Quanhong;Zhang, Jing
客观的。声动力疗法（SDT）是一种新颖且有前途的癌症疗法，结合使用超声波和血卟啉，可以诱导某些癌细胞凋亡。然而，SDT 诱导细胞凋亡的机制尚不清楚。本研究调查了 SDT 诱导的肉瘤 180 细胞凋亡。方法。在血卟啉存在下，用 1.75 MHz 连续聚焦超声处理细胞悬液 3 分钟，并通过流式细胞术、扫描电子显微镜、末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸-生物素缺口末端标记、共聚焦显微镜和凋亡相关蛋白分析来评估细胞凋亡。结果。 SDT后1小时观察到DNA断裂、凋亡小体和裂解的聚（三磷酸腺苷-核糖）聚合酶通过使用激光扫描共聚焦显微镜，我们发现Fas相关的死亡结构域和caspase 8从细胞质易位到质膜。通过免疫组化检测caspase 8和caspase 3的活性，结果表明SDT导致caspase 8激活，进而激活下游caspase 3。此外，使用caspase 8特异性抑制剂Z-Ile-Glu-Thr-Asp-氟甲基酮来证实caspase 8在细胞凋亡中的作用。结论。我们的数据主要表明SDT可以在体外诱导肉瘤180细胞凋亡，并且caspase 8可能在SDT诱导的细胞凋亡中发挥重要作用。
Objective. Sonodynamic therapy (SDT), a novel and promising cancer therapy that uses a combination of ultrasound and hematoporphyrin, can induce apoptosis in some cancer cells. However, the mechanism(s) of SDT-induced cell apoptosis is not well understood. This study investigated SDT-induced apoptosis in sarcoma 180 cells. Methods. Cell suspension were treated by 1.75-MHz continuous focused ultrasound in the presence of hematoporphyrin for 3 minutes, and apoptosis was assessed by flow cytometry, scanning electron microscopy, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling, confocal microscopy, and apoptosis-related protein analysis. Results. DNA breaks, apoptotic bodies, and cleaved poly (adenosine triphosphate-ribose) polymerase were observed 1 hour after SDT By using laser-scanning confocal microscopy, we found that the Fas-associated death domain and caspase 8 translocated from the cytoplasm to the plasma membrane. Activities of caspase 8 and caspase 3 were detected by an immunohistochemical assay The results suggested that SDT led to activation of caspase 8, which in turn activated downstream caspase 3. In addition, Z-Ile-Glu-Thr-Asp-fluoromethylketone, a specific inhibitor for caspase 8, was used to confirm the effect of caspase 8 in apoptosis. Conclusions. Our data primarily show that SDT can induce apoptosis in sarcoma 180 cells in vitro, and caspase 8 may play an important role in SDT-induced apoptosis.