肝素酶介导的裸鼠肝癌门静脉微瘤栓形成  

Heparanase-mediated formation of portal venous microemboli in nude mice of hepatocellular carcinoma

作　　者：陈晓鹏[1] 岳朝辅 卢林明[2] 张文君[1] 

机构地区：[1]皖南医学院弋矶山医院肝胆外科,安徽芜湖241001 [2]皖南医学院弋矶山医院病理科,安徽芜湖241001

出　　处：《第三军医大学学报》2016年第21期2297-2302,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81272412)~~

摘　　要：目的探寻肝癌细胞中高表达的肝素酶(heparanase,HPSE)对裸鼠肝癌门静脉微瘤栓形成的影响。方法根据既往研究结果,分别选取高表达和相对较低表达HPSE的HCCLM3和Hep G2两种人肝癌细胞;调整细胞浓度,行裸鼠下腹腔注射3×106个癌细胞(均300μL);至第3周和第5周时,分别取标本观察网膜和肝脏成瘤率、血管侵犯和门静脉微瘤栓形成情况,以建立肝癌微瘤栓模型和普通肝癌模型。模型鉴定并复制,然后于腹腔注射癌细胞后第2周开始分别皮下注射250 IU的HPSE抑制剂肝素钠,1次/3 d,共2周,并设生理盐水作为对照;仍于细胞注射第3周和第5周时,观察上述指标。结果裸鼠腹腔接种后一般情况无改变;第3周时体质量明显增加(P<0.05)。第3周和第5周时,两种肝癌细胞网膜成瘤率均为5/5。第3周时,HCCLM3细胞肝脏成瘤率(2/5)高于Hep G2细胞(0/5)(P<0.05);第5周时二者肝脏成瘤率相当(5/5 vs 4/5),但HCCLM3细胞肝转移灶体积较大。第3周和第5周时,HCCLM3细胞网膜血管(或门静脉)侵犯比例(2/5,5/5)均高于Hep G2细胞(0/5,3/5,P<0.05);第3周,两种癌细胞门静脉微瘤栓发生率相当(1/5 vs 0/5);第5周时,HCCLM3细胞门静脉微瘤栓发生率(5/5)显著高于Hep G2细胞(0/5,P<0.05)。应用肝素钠皮下注射至接种第3周时,HCCLM3细胞网膜及肝脏转移、血管侵犯及微瘤栓形成显著少于生理盐水组(P<0.05),Hep G2细胞网膜成瘤亦减少(P<0.05)。第5周时,两种细胞均再现肝转移灶,且HCCLM3细胞复现门静脉微瘤栓,但肝素组HCCLM3细胞所致的网膜和肝脏成瘤、血管侵犯和门静脉微瘤栓形成率仍明显少于生理盐水组(P<0.05)。结论腹腔注射高表达HPSE的HCCLM3细胞可成功介导肝癌门静脉微瘤栓形成;HPSE可能是微瘤栓形成重要因素。Objective To determine the effect of high expression of heparanase（ HPSE） in hepatocellular carcinoma（ HCC） cells on the formation of portal venous microemboli（ PVME） in HCC nude mice. Methods HCCLM3 cells with high expression of HPSE and Hep G2 cells with relatively low expression of HPSE were selected basing on our previous study. Nude mice were intraperitoneally injected with 300 μL of HCC cells（ 3 × 106cells）,and the tumor formation in the omentum and liver were respectively observed in 3and 5 weeks after injection. The vessel invasion and PVME were observed by microscopy. After the HCC models were established,the nude mice were subcutaneously injected with heparin sodium at a dose of 250 IU once per 3 d for 2 weeks in 2 weeks after HCC cells injection,and normal sodium was used for control.Results All nude mice survived without obvious changes in diet and activity,and their weight significantly increased in the 3rd week（ P〈0. 05）. For the 2 kinds of HCC cells,the tumor formation rate was both 5/5in the omentum at the 3rd and 5th weeks. The tumor formation rate of HCCLM3 cells in the liver was significantly higher than that of Hep G2 cells at the 3rd week（ 2/5 vs 0/5,P〈0. 05）,and the rates of the 2kinds of cells were similar at the 5th week（ 5/5 vs 4/5）,but the tumor sizes were larger in those induced by HCCLM3 cells. At the the 3rd and 5th weeks,HCCLM3 cells mediated more vessel invasions in the omentum and portal vein than Hep G2 cells（ 2/5 vs 0/5,5/5 vs 3/5,both P〈0. 05）. PVME rate was similar in the HCCLM3 cells and Hep G2 cells at the 3rd week（ 1/5 vs 0/5）,but significantly higher in the former than the latter cells at the 5th week（ 5/5 vs 0/5,P〈0. 05）. Subcutaneous injection of heparin for 2 weeks attenuated the omentum and liver metastases,vessel invasion and PVME in the HCCLM3 cells group compared with normal sodium group at the 3rd week（ all P〈0. 05）. Hep G2 cells induced omentum metastasis was also decreased obviously（ P〈0. 05）. After hep

关 键 词：肝素酶 肝细胞癌 门静脉 瘤栓 血管侵犯 

分 类 号：R-332[医药卫生] R345.72