生存素慢病毒载体局部治疗大鼠胫骨骨肉瘤  

Observation of survivin lentiviral vectorin tibia local curative effect on the treatment of osteosarcoma in rats

作　　者：孙保勇[1] 李敏[1] 王方欣 杨明山[1] 

机构地区：[1]山东省肿瘤医院骨与软组织肿瘤科,济南250117

出　　处：《中华实验外科杂志》2015年第8期1950-1952,共3页Chinese Journal of Experimental Surgery

摘　　要：目的建立生存素（Survivin）基因慢病毒载体和大鼠皮下骨肉瘤动物模型，观察Survivin基因慢病毒载体局部治疗大鼠皮下骨肉瘤的疗效。方法选取60只大鼠，采用注射法将骨肉瘤细胞种植在裸鼠下肢胫骨骨髓腔内，每天注射1次，连续注射4周，建立大鼠皮下骨肉瘤动物模型，将骨肉瘤模型裸鼠随机分为Survivin基因慢病毒载体组、空白载体组和空白对照组，分别于胫骨骨肉瘤局部注射Survivin一小干扰RNA（siRNA）的慢病毒载体50斗l、空白病毒载体和生理盐水各1次，4周后观察抑瘤率。分别于治疗前、治疗后（第2、7、14、28天）取原位肿瘤，行病理切片检查，同时采大鼠模型的血清检测其碱性磷酸酶（ALP）水平。结果（1）肿瘤生长曲线示慢病毒载体组骨肉瘤生长速度明显慢于空白对照组和空白载体组（P〈0．05），慢病毒载体组、空白载体组抑瘤率分别为45．12％、8．81％；慢病毒载体组肿瘤体积明显小于空白对照组和空白载体组（P〈0．01）。（2）圯镜下：Survivin基因慢病毒载体治疗后2d可见骨肉瘤局部形态尚保持完整，瘤区表皮呈暗紫色。卜肢水肿较前减轻。治疗7d后骨肉瘤坏死脱落，局部红肿充血带明显消退。治疗14d后，大鼠胫骨骨肉瘤坏死组织已开始修复，骨肉瘤边缘坏死处有结痂。（3）电镜下：Survivin基因慢病毒载体治疗前，大鼠胫骨骨肉瘤细胞大小不一，大量异形核，核膜不规则，异染色质边集，核浆比增大。经Survivin基因慢病毒载体治疗骨肉瘤14d后，可见骨肉瘤细胞破坏明显，细胞核膜不完整，核破裂或溶解。（4）骨肉瘤大鼠模型，AIJP水平与未治疗组比较差异有统计学意义（P〈0．01）。经过Survivin基冈慢病毒载体治疗后的大鼠模型，ALP水平有所降低，但直到28d后，其ALP水平仍高于正常水平（P〈O．05）。结论慢病毒载体介导的SurObjective To establish survivin gene lentiviral vector and subcutaneous osteosarcoma in rats model, observe the treatment effect of survivin lentiviral vector in rats with subcutaneous osteosarco- ma. Methods 60 rats were selected, the injection ofosteosarcoma cellsgrown in nude mice bone marrow cavity of tibia, 1 daily injections, continuous injection for 4 weeks, rat subcutaneous osteosarcoma rats model, the model of osteosarcoma in nude mice were randomly divided into survivin lentiviral vector group, blankplasmid group and blank control group. Tibial osteosarcoma, respectively in the local injection of survivin -small interfering RNA （siRNA） lentiviral vector 50 Ixl, blank vector and normal saline respec- tively for 1 times, for 4 weeks to observe the inhibition rate of tumor. Respectively before treatment, after treatment （2, 7, 14, 28 d） and in situ tumor, pathological biopsy, serum and rat model of detection of al- kaline phosphatase （ALP） level. Results （1） The tumor growth curve showed thatthe lentiviral vector group, tumor growth was slower than that of the blank control group and empty vector group （ P 〈 0. 05 ）, lentivirus vector group, blank vector group, the inhibition rates were 45.12%, 8.81% ; lentivirus vector group tumor volume was significantly less than the control groupand the blank vector group （ P 〈 0. 01 ）. （ 2 ） Light microscope ： the survivin lentiviral vector of osteosarcoma is still visible local shape intact tumor skin rafter treatment 2 days, dark purple, lower extremity edema relieved. After treatment 7 days, in osteo- sarcoma necrosis, local swelling and hyperemia with significant regression. After treatment 14 days, in a rat model of tibial osteosarcoma tissue necrosis has been repaired, scabby osteosarcoma necrosis. （3） Elec- tron microscope ： survivin lentiviral vector before treatment, in rats model of tibial osteosarcoma cell size, a large number of irregular nucleus, irregular nuclear membrane, heterochromatin margination, nuclear cyto-

关 键 词：骨肉瘤 生存素 慢病毒载体 

分 类 号：R394[医药卫生—医学遗传学]