| 摘要: |
| 目的：观察缺血再灌注损伤对裸鼠肝癌生长及癌旁组织转移复发相关基因（VEGF、MMP-9）表达的影响。方法：将人肝癌细胞Hep3B原位种植于裸鼠肝脏制备荷肝癌裸鼠模型，荷瘤裸鼠随机分为缺血再灌注后1 h、6 h、5 d、7 d及假手术组（n=8），缺血再灌注组荷肝癌裸鼠采用肝门阻断法制备缺血再灌注损伤模型，假手术组不阻断肝门血供，其余同缺血再灌注组。缺血再灌注后1 h、6 h检测裸鼠肝功能（ALT、AST）的变化（n=8）；实时荧光定量PCR检测瘤旁组织VEGF和MMP-9 mRNA的表达，并与正常对照组作比较（n=6）。缺血再灌注后5 d、7 d 取肝组织，H-E染色观察肝组织病理学变化，计算肿瘤周边变性坏死区域（n=6）。剩余荷瘤裸鼠于缺血再灌注后2周处死取肿瘤，测定缺血再灌注组及假手术组裸鼠肿瘤体积及质量。结果：术后2周缺血再灌注组肿瘤体积(mm3) 及质量(g) 明显高于假手术组\[（209.6±25.74）vs（330.6±32.01），（0.214±0.036） vs （0.374±0.045），P<0.01\]。缺血再灌注后1、6 h裸鼠血浆ALT和AST增高，明显高于假手术组（P<0.01）。H-E染色结果表明再灌注后5 d肿瘤周边组织炎症细胞浸润、变性坏死区域较假手术组更明显（P<0.05）；再灌注后7 d肿瘤周边变性坏死区域较5 d减少（P<0.05），但变性坏死区域被肿瘤细胞浸润代替。荧光定量结果表明，缺血再灌注后1 h和6 h癌旁组织VEGF和MMP-9 mRNA表达明显高于假手术组（P<0.01），且二者表达具有一定相关性（r=0.418,P<0.01）。结论：肝门阻断所导致的缺血再灌注损伤加速了裸鼠肝肿瘤细胞的生长，促进了癌旁组织中与转移复发相关基因（VEGF、MMP-9）的表达。 |
| 关键词: 肝肿瘤 缺血再灌注 VEGF MMP-9 |
| DOI：10.3724/SP.J.1008.2008.00495 |
| 投稿时间：2008-04-16修订日期：2008-04-26 |
| 基金项目:国家自然科学基金（30772512）. |
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| Ischemia-reperfusion promotes hepatic cancer growth and expression of VEGF and MMP-9 in adjacent tissues of cancer in nude mice |
| AI Li△, ZHANG Xiao-feng△,LIU Peng,CHEN Lei,HU He-ping* |
| (Department of Comprehensive Treatment, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438, China) |
| Abstract: |
| Objective：To observe the influence of ischemia-reperfusion (I/R) on hepatoma growth and on the expression of genes associated with tumor metastasis and recurrence (VEGF and MMP-9) in the adjacent tissues of cancer in nude mice. Methods: BALB/c nude mouse model bearing Hep3B-tumor in the liver was established and the model mice were evenly randomly into 5 groups: sham group and ischemia/reperfusion 1 h, 6 h, 5 d, and 7 d groups (n=8). I/R models were established by blocking porta hepatic; the sham group underwent the same treatment as the I/R model group except for blocking of porta hepatic. ALT and AST were detected in I/R 1 h and 6 h groups. Real-time-PCR was employed to detect the change of VEGF and MMP-9 in the adjacent tissues of cancer and the results were compared with that of the control group (n=6). Histopathological changes of liver were studied by H-E staining and necrotic areas were calculated in I/R 5 d and 7 d groups(n=6). The remnant tumor bearing mice were sacrificed 2 weeks after I/R to measure the volume and mass of the tumors. Results: Two weeks later, the tumor volume and mass in I/R group were increased compared with those in the sham group (\[209.6±25.74\] mm3 vs \[330.6±32.01\] mm3,\[0.214±0.036\] g vs \[0.374±0.045\] g, P<0.01). Levels of ALT and AST were significantly elevated in I/R 1 h and 6 h groups compared with those in the sham group (P<0.01). H-E staining showed that the infiltration of inflammatory cells around the tumors and the areas of necrosis became more prominent 5 days after I/R compared with the sham operation group (P<0.05). The necrosis area was reduced 7 days after I/R compared with 5 days after I/R (P<0.05); however, the necrosis area was replaced by invasion of tumor cells (P<0.05). It was found that the expression of VEGF and MMP-9 mRNA was higher in the adjacent tissues of cancer after I/R than that in the sham group (P<0.01), and the expression of the 2 was correlated with each other (r=0.418, P<0.01). Conclusion: I/R induced by blocking porta hepatic can accelerate tumor cell growth and promote expression of VEGF and MMP-9 in the adjacent tissues of cancer. |
| Key words: liver cancer ischemia/reperfusion VEGF MMP-9 |
