消炎痛抑制肝细胞肝癌生长转移的研究

Effects of indomethacin on the growth and metastasis of hepatocellular carcinoma

目的观察消炎痛（Indomethacin）对有转移潜能的人肝癌MHCC97L细胞增殖侵袭的影响和对裸鼠肝癌生长和转移的抑制作用。方法（1）体外实验：采用0．2mmo]／L的消炎痛分别作用于MHCC97L细胞，观察细胞增殖、侵袭实验、运动实验和血管内皮生长因子（VEGF）和基质金属蛋白酶2（MMP-2）蛋白表达[酶联免疫吸附试验（ELISA）]。（2）体内实验：建立转移性人肝癌裸鼠原位模型后，将裸鼠随机分为对照组和消炎痛组。6周后处死动物，测量肿瘤体积，计算抑瘤率、肺转移灶数目及肺转移率。免疫组织化学方法检测VEGF、MMP-2、环氧合酶-2（COX-2）蛋白的表达。结果（1）体外实验：0．2mmol／L消炎痛明显抑制MHCC97L细胞增殖（P值均〈0．01），消炎痛组穿过人工基底膜（侵袭实验）和上室底膜（运动实验）的细胞数分别为2．2±1．3和4．4±1．1，明显低于对照组（11．4±1．9和12．8±1．8，P值均〈0．01）；ELISA法检测发现，消炎痛组VEGF蛋白和MMP-2蛋白含量和对照组比较明显降低（P值均〈0．01）。（2）体内实验：对照组、消炎痛组肿瘤体积分别为（1700±422）mm3和（1170±585）mm。（P〈0．05），肺转移率分别为75％和50％（P〉0．05），平均肺转移灶数目分别为2．92±2．07和1．33±1．56（P〈0．05）；与对照组比较，消炎痛组的抑瘤率为31．2％。免疫组织化学染色显示，消炎痛组VEGF、MMP-2、COX-2蛋白的表达和对照组比较均有降低（P值均〈0．01）。结论在-定条件下，消炎痛可抑制肝细胞肝癌的生长转移，其作用和抑制VEGF蛋白和MMP-2蛋白的表达有关。

Objective To study the effects of indomethacin on proliferation and invasion of hepa- tocellular carcinoma （HCC） cell line MHCC97L with metastatic potential and the effect of indomethacin on the growth and metastasis of HCC. Methods （ 1 ） In vitro ： Proliferation, Transwell invasion assay, cell motility assay, vascular endothelial growth factor （VEGF） and matrix metalloproteinase-2 （MMP-2） protein activity were evaluated after cells were treated with 0. 2 mmol/L indomethacin. （2） In vivo ： Mice bearing xenografts in the liver were randomly divided into control and indomethacin groups. At the end of sixth week, the mice were killed and tumor volume, inhibitory rate, immunohistochemistry assay （IHA） and metastasis were evaluated. Results （ 1 ） In vitro ： 0. 2 mmol/L indomethacin could inhibit the proliferation of MHCC97L cells markedly （P 〈 0. 01 ）. The average amount of invading cells per field in cell invasion assay and motility assay was 2. 2 ± 1.3 and 4. 4 ± 1.1 respectively in indomethacin group, significantly less than in control group （ 11.4 ± 1.9 and 12. 8 ± 1.8 respectively, P 〈0. 01 ）. The expression of VEGF and MMP-2 in cells treated with indomethacin was significantly lower than in control group （P 〈 0. 01 ）. （2）In vivo： Tumor volume, incidence and number of lung metastases in control and indomethacin groups were （1700±422） mm3 and （1170±585） mm3（p〈0.05）, 75% and 50% （P〉0.05）, 2.92 ±2.07 and 1.33 ± 1.56 （P 〈 0. 05 ）, respectively. Inhibition rate in indomethacin group was 31.2%. IHA showed that the expression of VEGF, MMP-2, and cyclooxygenase-2 （COX-2） was down-regulated in indomethacin group （ P 〈 0. 01 ）. Conclusion Indomethacin could inhibit the growth and metastasis of HCC, which was in part mediated by down-regulation of VEGF and MMP-2.