转染TRAIL的内皮祖细胞对人卵巢癌裸鼠皮下移植瘤的抑制作用

Antitumor Effect of Endothelial Progenitor Cells with TRAIL Gene Transfection on Ovarian Carcinoma Xenografts in Nude Mice

背景与目的:肿瘤坏死因子相关凋亡诱导配体(tumornecrosisfactor-relatedapoptosisinducingligand,TRAIL)有广谱的抗瘤作用,且对正常组织细胞无毒性,因此有望应用于肿瘤基因治疗。内皮祖细胞(endothelialprogenitorcells,EPCs)在体内能定向归巢于肿瘤,参与肿瘤新生血管的建立。本研究以EPC为载体,观察TRAIL转染EPCs对人卵巢上皮癌裸鼠皮下移植瘤的治疗作用。方法:用磁珠分离法从脐血中分离EPCs,并进行体外培养扩增。用脂质体将带有GFP-TRAIL基因的质粒转入EPCs(TRAIL-EPCs)。将转染后的EPC经尾静脉注入3AO卵巢癌裸鼠皮下移植瘤模型。流式细胞仪检测各组中绿色荧光蛋白(Green-Fluoroprotein,GFP)表达情况,观察各组移植瘤体积的变化,计算抑瘤率。结果:静脉注射转染TRAIL后的EPC,对卵巢上皮癌裸鼠皮下移植瘤生长具有明显抑制作用,对照组裸鼠的瘤重(0.226±0.209)g,而TRAIL细胞因子治疗组、GFP-TRAIL转染组裸小鼠的瘤重分别为(0.118±0.164)g、(0.075±0.084)g;TRAIL细胞因子组的抑瘤率为48.1%,TRAIL转染组抑瘤率为66.9%。肿瘤组织石蜡切片HE染色检查显示TRAIL细胞因子组,TRAIL转染组转对照组有更多的出血坏死区。TRAIL细胞因子组,TRAIL转染组均无明显毒副作用的表现。结论:TRAIL细胞因子和TRAIL-EPCs对人卵巢癌裸鼠皮下移植瘤均有明显的抑制作用,EPC在裸鼠皮下移植瘤模型体内有一定的导向作用,有希望成为基因治疗的载体。

BACKGROUND ＆ OBJECTIVE- Tumor necrosis factorrelated apoptosis inducing ligand （TRAIL） can induce apoptosis in various cancer cell lines with little toxicity toward normal cells. It offers a promising therapeutic method against ovarian cancer. Endothelial progenitor cells （EPCs） could home to tumor lesion, and take part in angiogenesis. This study was to observe antitumor effect of EPCs with TRAIL gene transfection on human ovarian epithelial cancer xenografts in nude mice. METHODS. EPCs were isolated from human cord blood by magnetic bead selection, and cultured and amplified in vitro. Human ovarian epithelial cancer cell line 3AO cells were transplanted subcutaneously in immunodeficiency nude mice. EPCs were transfected with hTRAIL plasmid （TRAIL-EPCs）. TRAIL-EPCs, TRAIL, or culture media IMDM （control） was injected into the mice model of 3AO xenograft by caudal vein. The changes of tumor volume were observed, and the tumor growth inhibition rate was calculated. RESULTS： EPCs with TRAIL gene transfection obviously inhibited the growth of 3AO xenograft in nude mice; the tumor weight of control, TRAIL, and TRAIL-EPCs groups were （0.226±0.209） g, （0.118±0.164） g, and （0.075±0.084） g, respectively, the maximum tumor growth inhibition rate was significantly higher in TRAIL-EPCs group than in TRAIL group （66.9% vs. 48.1%, P〈0.05）. HE staining showed that there were more hemorrhage and necrosis regions in TRAIL and TRAIL- EPCs groups than in control group. No severe toxic reaction was observed in the 2 groups. CONCLUSIONS： Both TRAIL and TRAIL-EPCs can inhibit the growth of ovarian epithelial carcinoma xenograft in nude mice. EPCs have certain guidance effect in tumor xenografts in nude mice.