| 宁周雨,王 鹏,陈 颢.不可逆电穿孔治疗肝癌的临床前动物实验研究[J].肿瘤学杂志,2016,22(1):17-23. |
| 不可逆电穿孔治疗肝癌的临床前动物实验研究 |
| Pre-clinical Experimental Study of Animal Irreversible Electroporation Treatment for Liver Cancer |
| 投稿时间：2015-04-16 |
| DOI：10.11735/j.issn.1671-170X.2016.01.B004 |
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| 中文关键词: 不可逆电穿孔（IRE） 肝肿瘤 移植瘤 细胞凋亡 免疫功能 安全性 |
| 英文关键词:irreversible electroporation (IRE) liver neoplasms xenograft cell apoptosis immunity security. |
| 基金项目: |
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| 中文摘要: |
| 摘 要：［目的］ 通过研究肝脏移植瘤不可逆电穿孔（Irreversible Electroporation，IRE）术后细胞凋亡情况，观察IRE技术对肝脏移植瘤的抗肿瘤效果及安全性，为临床应用提供实验依据。［方法］ 12只新西兰大白兔肝脏移植瘤模型随机分为IRE治疗组和阴性对照组。治疗组对肝脏移植瘤给予IRE治疗，阴性对照组不予处理。治疗后第1d、3d、5d、7d、14d各处死2只，取血清进行肝肾功能、淀粉酶及肌酸激酶检测，ELISA方法检测血浆肌钙蛋白I（CTnI）、Caspase-3、TNF-α、VEGF因子水平。TUNEL法检测肿瘤组织细胞凋亡。免疫组织化学方法检测Bcl-2、HSP70和VEGF在肝脏移植瘤中的表达。［结果］ 治疗过程中，新西兰大白兔出现脉冲式全身震颤，但可控。术后血清学检测结果显示治疗后新西兰大白兔除肌酸激酶水平明显升高外，血清淀粉酶、肝肾功能均正常。血浆ELISA结果显示IRE有效地增加机体Caspase-3、TNF-α的分泌，同时降低VEGF表达，而且CTnI水平未见明显异常升高。HE染色显示治疗组坏死区域和正常区域边界清晰，坏死边缘区可见大量红细胞以及炎症细胞浸润，治疗后14d治疗区边缘出现肿瘤再生迹象。TUNEL法结果显示，与阴性对照组相比，IRE治疗不同时间组可以明显增加肝脏移植瘤组织凋亡。免疫组化结果表明，IRE治疗能够降低Bcl-2以及VEGF的表达，同时增加HSP70在移植瘤中的表达，增强IRE的抗肿瘤效应。［结论］ IRE能够通过诱导细胞凋亡、产生特异的抗肿瘤免疫效应及抑制血管生成来抑制肝脏肿瘤的生长，治疗方法安全、有效，为其进一步的临床应用提供了动物实验依据。 |
| 英文摘要: |
| Abstrat：［Purpose］To investigate the apoptosis of liver cancer after irreversible electroporation (IRE) operation，and to observe the antitumor effect and security of IRE technique for liver cancer xenograft，to provide experimental evidence for the clinical usage. ［Methods］ Twelve New Zealand white rabbit with liver cancer xenograft models were randomly divided into treatment group and control group. The treatment group received IRE，the control group received no treatment. Two rabbits in each group were executed on the 1st，3rd，5th，7th，14th day after IRE. Liver and kidney function and creatine kinase were tested，ELISA method was used to detect CTnI，Caspase-3，TNF-α and VEGF levels in the plasma. TUNEL assay was used to detect the apoptosis of tumor tissue. Immunohistochemistry was applied to detect Bcl-2，HSP70 and VEGF expressions in liver cancer xenograft. ［Results］ During the treatment，the rabbits had pulse-type tremor，but it was controllable. Post-treatment serological test showed that serum amylase，liver and kidney functions were within the normal range except significant higher level of creatine kinase. ELISA detection of plasma showed IRE increasing secretion of Caspase-3， and TNF-α，while reducing VEGF expression，and no elevation of CTnI. HE staining showed a clear borderline between the necrotic and normal areas，with a large number of red blood cells and inflammatory cell infiltration on the edge. After 14 days of treatment，regeneration signs appeared in the target tumor tissues. TUNEL assay showed that IRE significantly increased the apoptosis in liver cancer xenograft cells compared with the control group. Immunohistochemistry results showed that IRE decreased the expressions of Bcl-2 and VEGF，increased HSP70 expression in liver cancer xenograft，and improved anti-tumor effect of IRE. ［Conclusion］ IRE might inhibit the growth of liver tumor via inhibiting angiogenesis，inducing apoptosis and producing specific anti-tumor immune response. IRE is a safe and effective approach for liver cancer. With the evidence from the experiment，this study may support further clinical IRE application. |
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