Institute of Ultrasound Imaging, Chongqing Medical University, Chongqing, China.
J Ultrasound Med. 2009 Dec;28(12):1695-702. doi: 10.7863/jum.2009.28.12.1695.
Osteosarcoma is one of the most common malignant cancers afflicting young adults. Ultrasound is a new therapeutic modality for controlling malignant cancers. Enhancing the efficacy of ultrasound treatment will improve clinical outcomes. The aim of this study was to investigate the killing action of ultrasound on osteosarcoma enhanced by hematoporphyrin monomethyl ether (HMME) in vivo.
An animal model of an osteosarcoma xenograft was set up to investigate the inhibitory effect of sonoactivating HMME on osteosarcoma. High-performance liquid chromatography was used to analyze the time course of HMME in the osteosarcoma xenograft. Three hours after intravenous (IV) administration of HMME, ultrasound radiation was administered in the osteosarcoma xenograft. On day 7 after ultrasound radiation, the tumor volume and weight were measured and calculated for effect assessment, hematoxylin-eosin staining for histopathologic examination, immunohistochemical staining for proliferating cell nuclear antigen (PCNA) expression, and terminal deoxyuridine nick end-labeling (TUNEL) staining for apoptosis examinations.
The peak value of HMME in osteosarcoma tissues increased with time after IV administration of HMME and reached a plateau at 3 hours. The increasing rates of the tumor volume and weight in the control group were very fast, but the increasing rates in the ultrasound-alone group were slower, and those in the ultrasound-HMME group were the slowest throughout the observation period. There was a significant difference between the HMME-ultrasound, ultrasound-alone, and control groups (P < .01). Hematoxylin-eosin staining showed that some cells had typical cell death such as pyknosis and nuclear fragmentation after ultrasound radiation alone. More cells with pyknosis, nuclear fragmentation, and even karyolysis were found after HMME-ultrasound treatment. Immunohistochemical staining showed that the percentage of PCNA-positive cells decreased and that of TUNEL-positive cells increased after ultrasound treatment alone, and the changes in the PCNA- and TUNEL-positive percentages were significantly enhanced by pretreatment with HMME (20 mg/kg, IV) for 3 hours and ultrasound radiation (10.5 MHz) for 120 seconds at an intensity of 0.8 W/cm(2) (P < .05).
Hematoporphyrin monomethyl ether pretreatment could substantially enhance the growth inhibition of ultrasound on osteosarcoma, which suggests that HMME is an efficient sonosensitizer, and ultrasound radiation with HMME could be developed as a new modality for treating osteosarcoma.
骨肉瘤是最常见的侵袭青年人群的恶性肿瘤之一。超声是一种治疗恶性肿瘤的新方法。增强超声治疗的疗效将改善临床结果。本研究的目的是研究血卟啉单甲醚（HMME）增强的超声对骨肉瘤的杀伤作用。
建立骨肉瘤异种移植动物模型，研究声激活 HMME 对骨肉瘤的抑制作用。采用高效液相色谱法分析骨肉瘤异种移植中 HMME 的时间过程。静脉（IV）给予 HMME 后 3 小时，在骨肉瘤异种移植中给予超声辐射。超声辐射后第 7 天，测量和计算肿瘤体积和重量进行效果评估，进行苏木精-伊红染色进行组织病理学检查，进行增殖细胞核抗原（PCNA）表达的免疫组织化学染色，以及末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色进行凋亡检查。
IV 给予 HMME 后，骨肉瘤组织中 HMME 的峰值随时间增加，在 3 小时达到平台期。对照组肿瘤体积和重量的增长率非常快，但超声组的增长率较慢，整个观察期超声-HMME 组的增长率最慢。HMME-超声、超声单独和对照组之间存在显著差异（P <.01）。苏木精-伊红染色显示，单独超声辐射后，一些细胞出现典型的细胞死亡，如固缩和核碎裂。用 HMME-超声处理后，发现更多的细胞出现固缩、核碎裂，甚至核溶解。免疫组织化学染色显示，单独超声处理后 PCNA 阳性细胞的百分比下降，TUNEL 阳性细胞的百分比增加，HMME（20mg/kg，IV）预处理 3 小时和超声辐射（10.5MHz）120 秒后，PCNA 和 TUNEL 阳性细胞百分比的变化明显增强，强度为 0.8W/cm(2)（P <.05）。
HMME 预处理可显著增强超声对骨肉瘤的生长抑制作用，表明 HMME 是一种有效的声敏剂，HMME 增强的超声辐射可能成为治疗骨肉瘤的一种新方法。
