NK4修饰骨髓间充质干细胞靶向治疗裸鼠肝癌  

Targeted therapy of hepatocellular carcinoma via systemic injection of mesenchymal stem cells mod- ified by NK4 in nude mice

作　　者：李光林 朱金海[1] 陈燕凌[1] 

机构地区：[1]福建医科大学附属协和医院肝胆外科,福州350001

出　　处：《中华实验外科杂志》2017年第10期1677-1679,共3页Chinese Journal of Experimental Surgery

基　　金：福建省科技计划重点项目（2014Y0057）;福建省自然科学基金（2014J01323）

摘　　要：目的探讨肝细胞生长因子拮抗剂NK4基因修饰骨髓间充质干细胞（MSCs）对裸鼠肝癌的治疗作用。方法体外构建NK4慢病毒载体并感染MSCs，获得稳定表达NK4的MSCs（LV-NK4-MSCs），Western blot与酶联免疫吸附试验（ELISA）分析其NK4的表达。将LV-NK4-MSCs上清液作用于3组肝癌细胞SMMC-7721、MHCC-97H、HepG2，观察其对肝癌细胞增殖和凋亡的影响。建立裸鼠肝癌皮下移植瘤模型，尾静脉注射LV-NK4-MSCs进行治疗，观察MSCs在体内迁移情况，测量荷瘤裸鼠的肿瘤体积、生存时间，ELISA检测瘤内及血浆中NK4浓度、免疫组织化学检测CD34、血管内皮生长因子（VEGF）、细胞核增殖抗原（Ki-67）表达情况。 结果成功获得稳定表达NK4的LV-NK4-MSCs，能够表达和分泌NK4。NK4能够明显抑制3组肝癌细胞的增殖，促进凋亡形成。在裸鼠肝癌模型中，MSCs能靶向迁移至肝癌组织中，而几乎不出现在正常组织中。与对照组比较，LV-NK4-MSCs治疗组的瘤体内NK4水平明显高表达[（379.80±16.83） pg/ml比（29.68±5.59） pg/ml，P＝0.021]，肿瘤体积显著缩小[（211.40±14.13） mm3比（457.70±93.74） mm3，P＝0.009]，CD34、VEGF及Ki-67表达显著降低，生存时间明显延长。结论NK4修饰MSCs能够抑制肝癌细胞的增殖，能在体内靶向迁移至裸鼠肝癌组织中，抑制裸鼠肝癌的生长，延长生存期。ObjectiveTo investigate anti-tumor effect of mesenchymal stem cells （MSCs） modified by NK4 on hepatocellular carcinoma.MethodsMSCs modified by NK4 （LV-NK4-MSCs） were conducted by lentiviral transduction. Western blotting and enzyme linked immunosorbent assay （ELISA） were performed to analyze the expression level of NK4. SMMC-7721, MHCC-97H and HepG2 were supplemented with supernatant of LV-NK4-MSCs to observe the effect of NK4 on proliferation and apoptosis of hepatocellular carcinoma （HCC） cell lines. SMMC-7721 were used to establish an HCC subcutaneous mouse model. LV-NK4-MSCs were injected into tumor-bearing mice to visualize the distribution of MSCs in vivo. Tumor volume and survival time were monitored. ELISA was performed to detect the intratumoral level and plasma level of NK4. The expression of CD34, proliferation cell nuclear antigen （Ki-67） and vascular endothelial growth factor （VEGF） was analyzed by immunohistochemistry assay. ResultsLV-NK4-MSCs were conducted by lentiviral transduction and could secrete NK4 effectively. NK4 could inhibit the proliferation of three hepatocellular carcinoma （HCC） cells and promote apoptosis. MSCs mainly migrated to tumor site, rather than normal tissues. The intratumoral NK4 level in LV-NK4-MSCs group was higher than that in control group [（379.80±16.83） pg/ml vs. （29.68±5.59） pg/ml, P=0.021]. Tumor volume and the expression of CD34, Ki-67 and VEGF in the mice treated with LV-NK4-MSCs were significantly decreased than that in the control group [（211.40±14.13） mm3 vs. （457.70±93.74） mm3,P=0.009]. The median survival of mice treated with LV-NK4-MSCs was higher than that in the control group.ConclusionMSCs modified by NK4 could inhibit the proliferation of HCC cells. LV-NK4-MSCs could migrate to the tumor site in vivo, inhibit tumor growth and prolong survival.

关 键 词：NK4 肝癌 骨髓间充质干细胞 裸鼠 

分 类 号：R735.7[医药卫生—肿瘤]