塞来昔布抑制肝动脉断流术后增强的肝癌转移潜能

Significance of celecoxib on enhanced metastatic potential due to hepatic artery occlusion in hepato- cellular carcinoma

目的观察肝动脉断流后肝癌局部或机体全身的炎症反应，以及非甾体类抗炎药物塞来昔布（Celecoxib）对断流术后残癌恶性潜能的影响。方法建立具有自发转移潜能的肝癌原位移植＋肝动脉结扎（HAL）模型。酶联免疫吸附试验（ELISA）检测HAL对荷瘤动物血清内炎症因子肿瘤坏死因子（TNF）-A、白细胞介素（IL）_6、肝细胞生长因子（HGF）表达的影响；免疫组织化学分析肝脏原发瘤及转移靶器官（肺部）的炎症反应及环氧合酶-2（COX-2）表达；继而不同剂量的Celecoxib协同HAL处理荷瘤裸鼠，观察对上述炎症反应及转移的影响。结果HAL显著上调荷瘤动物血清中TNF-a、IL-6、HGF表达（3．78、2．91、1．82倍，P〈0．05），增加肝脏原发瘤局部坏死[HAL组（51．27±19．39）％比假手术组（19．69±5．49）％，P〈0．01]及组织内COX-2表达，引起转移靶器官（肺）明显炎症改变。协同使用高剂量Celecoxib（每天50mg／kg）下调组织内COX-2表达，进一步减少移植瘤体积[（1091．81±870．14）mm3比（2735．06±474．97）mm3，P〈0．01]，并显著抑制肿瘤肺转移（0／6比8／10，P〈0．01），延长荷瘤裸鼠生存时间[（81．67±8．51）d比（60．17±5．42）d，P〈0．05]；小剂量Celecoxib（每天12．5ms／kg）虽不能抑制原发瘤生长及组织内COX-2表达，但减少了肝癌肺转移（2／6比8／10，P〉0．05）。结论肝动脉断流增加荷瘤裸鼠全身及原发瘤或转移靶器官局部炎症反应；抗炎药物Celecoxib抑制炎症反应，通过依赖或不依赖COX-2的机制阻止断流后增强的残癌侵袭、转移。

Objective To investigate the local or systemic inflammatory reaction after hepatic ar- tery ligation （ HAL）, and to explore the therapeutic effect of celecoxib on HAL-enhanced metastatic poten- tial of hepatocellular carcinoma （HCC）. Methods The spontaneous metastatic model in nude mice was established with MHCC97 ceils via orthotopic implantation and further to HAL. The proinflammatory cyto- kines such as tumor necrosis factor （TNF）-a, interleukin （IL）-6, and hepatocyte growth factor （HGF） in serum in HAL group were compared with those in sham-operation group by enzyme linked immunosorbent assay （ELISA）. The signs of inflammation in primary tumor or the lung from two groups were observed by immunohistochemistry and the expression of cyclooxgenase 2 （COX-2） in xenografts was evaluated. The in- fluence of celecoxib at different doses on tumor growth, pulmonary metastatic ratio, and COX-2 expression in tumor tissues due to HAL was investigated. Results The serum levels of proinflammatory cytokines （TNF-ot, IL-6 and HGF） were elevated after HAL （3.78 times; 2. 91 times; 1.82 times ,P 〈 0. 05）. Also the necrotic area in the xenograft as scored with hepatic replacement area （HRA） was increased by HAL [ HAL group： （51.27±19. 39） % vs. sham-operation group： （ 19.69±5.49） % ;P 〈0. 01 ]. Especially, the lungs of nude mice in the HAL group showed more inflammatory damage, characterized by intense inflammatory infiltration, thickening of alveolar walls, and severe vascular margination. As compared with normal saline （NS） treatment following HAL, HAL in combination with high dose of celecoxib （50 mg/kg every day） resulted in significantly reduced tumor size [ （1091.81±870. 14） mm3 vs. （2735.06±474. 97 ）mm3 ,P 〈0. 01 ] , decreased pulmonary metastasis （0/6 vs. 8/10 ,P 〈0.01 ）, and prolonged survival [ （81.67±8.51 ） days vs. （60. 17±5.42） days,P 〈0. 05 ]. HAL followed by celecoxib at a dose of 12.5 mg/kg every, day neither slowed down residual tumors growth nor prolonged survival, but inhibited the formation of pulmonary metastatic nodules though the difference （ compared with sham operation） was not statistically significant （ 2/6 vs. 8/10, P 〉 0. 05 ）. Interestingly, as shown in supplementary study, we did not detect the conspicuous changes of COX-2 expression in tumor tissues-derived HAL with low dose eele- coxib-treated mice, while the striking decrease of COX-2 expression was observed in treatments by HAL combined with moderate or high dose celeeoxib. Conclusion HLA leads to local or systemic inflammatory reaction, which is inhibited by celecoxib as accompanied with arrest of HAL-induced metastatic potential in residual HCC cells.