Experimental research on apoptosis induced following microwave coagulation therapy against liver cancer

微波凝固治疗肝癌诱导细胞凋亡的实验研究

• 批准号: 11671256
• 负责人: KAWANO Katsunori
• 金额: $ 0.7万
• 依托单位: OITA MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671256/
• 关键词: Microwave coagulation; Liver cancer; Apoptosis; Heat shock protein; Residual tumor

Microwave coagulation therapy (MCT), one of the local hyperthermia, has been widely performed to liver cancer patients, and yielded better clinical results than PEIT and TAE. In spite of the efficacy, the precise mechanism for expansion of cell death area following the therapy has still been unknown. The aim of this study was to understand the cellular mechanism of MCT focusing on whether the heat-induced apoptosis is occurred in the liver tissue by sequential examinations of caspase-3 activity and DNA fragmentation.One bout of MCT was applied to the rat liver using a monopolar needle electrode under setting of 30 W for 30 sec as coagulation time and 10 sec as dissociation time. The rats were sacrificed immediately, 2, 6, 12, 24, 72 and 168 h after MCT. The liver surface area was measured for evaluation of expansion of cell death area. For the assessment of the apoptosis, the TUNEL staining and the TUNEL positive cell counting was examined for the localization and the quantitiveness of DNA fragmentation. The caspase-3 activity in the specimen extracted from MCT site was measured for enzymatic evaluation of apoptosis.The caspase-3 activity showed four-fold increase from the value of 0 h, having a peak at 2 h after MCT. The DNA fragmentation significantly increased at 6 h, and decreased thereafter, assessed by the TUNEL positive cell count. The discolored liver surface area gradually increased until 12 h after MCT, sustained from 12 h to 72 h, and subsided thereafter.It was demonstrated following MCT that the apoptosis of hepatocytes was induced around the ablation area. This may explain the mechanism for the expansion of cell death area subsequent to MCT.

微波凝固治疗（MCT）作为局部热疗的一种，已被广泛应用于肝癌患者的治疗，其临床效果优于PEIT和TAE。尽管有疗效，但治疗后细胞死亡面积扩大的确切机制仍然未知。为探讨MCT的细胞机制，通过检测肝组织中caspase-3活性和DNA片段化的变化，探讨热诱导肝细胞凋亡的机制，采用单针电极，在30 W的功率下，对大鼠肝脏进行一次MCT，凝固时间30 s，解离时间10 s。在MCT后立即、2、6、12、24、72和168 h处死大鼠。测量肝表面积以评价细胞死亡面积的扩大。采用TUNEL染色和TUNEL阳性细胞计数法检测细胞凋亡的发生和DNA片段的定位及定量。从MCT部位提取的标本中测定caspase-3活性用于细胞凋亡的酶学评价，caspase-3活性从0 h的值增加了4倍，在MCT后2 h达到峰值。TUNEL阳性细胞计数显示，DNA片段化在6 h时显著增加，此后下降。MCT后12 h肝表面积逐渐增大，12 ~ 72 h持续，之后逐渐消退，表明MCT诱导了消融区周围肝细胞的凋亡。这可能解释了MCT后细胞死亡面积扩大的机制。

项目成果

Tsuyoshi Ohno, Katsunori Kawano, Atsushi Sasaki, Masanori Aramaki, Takanori Yoshida, Seigo Kitano: "Expansion of an ablated site and induction of apoptosis after microwave coagulation therapy in rat liver"J Hepatobiliary Pancreat Surg. 8 (3). 360-366 (200

Tsuyoshi Ohno、Katsunori Kawano、Atsushi Sasaki、Masanori Aramaki、Takanori Yoshida、Seigo Kitano：“大鼠肝脏微波凝固治疗后消融部位的扩大和细胞凋亡的诱导”J Hepatobiliary Pancreat Surg。

Ohno T, Kawano K, Sasaki A, Aramaki M, Yoshida T, Kitano S: "Expansion of an ablated site and induction of apoptosis'after microwave coagulation therapy in rat liver"Journal of Hepato-Biliary-Pancreatic Surgery. 8. 360-366 (2001)

Ohno T、Kawano K、Sasaki A、Aramaki M、Yoshida T、Kitano S：“大鼠肝脏微波凝固治疗后消融部位的扩大和细胞凋亡的诱导”《肝胆胰外科杂志》。

Tsuyoshi Ohno, Katsunori Kawano, Hirotaka Yokoyama, Kouichirou Tahara, Atsushi Sasaki, Masanori Aramaki, Seigo Kitano: "Microwave coagulation therapy accelerates growth of residual cancer in rat liver"J Hepatol. (in press).

Tsuyoshi Ohno、Katsunori Kawano、Hirotaka Yokoyama、Kouichirou Tahara、Atsushi Sasaki、Masanori Aramaki、Seigo Kitano：“微波凝固疗法加速大鼠肝脏中残留癌症的生长”J Hepatol。

Ohno T, Kawano K, Yakoyama H, Tahara K, Sasaki A, Aramaki M, Kitano S: "Microwave coagulation therapy accelerates growth of cancer in rat liver"Journal of Hepatology. (印刷中). (2002)

Ohno T、Kawano K、Yakoyama H、Tahara K、Sasaki A、Aramaki M、Kitano S：“微波凝固疗法加速大鼠肝脏中的癌症生长”肝脏病学杂志（2002 年）。

Ohno T, Kawano K, Yokoyama H, Tahara K, Sasaki A, Aramaki M, Kitano S: "Microwave coagulation therapy accelerates growth of cancer in rat liver"Journal of Hepatology. (印刷中). (2002)

Ohno T、Kawano K、Yokoyama H、Tahara K、Sasaki A、Aramaki M、Kitano S：“微波凝固疗法加速大鼠肝脏中的癌症生长”肝脏病学杂志（2002 年）。