血管抑素基因治疗人卵巢癌裸鼠皮下移植瘤的实验研究

目的:研究血管抑素(Angiostatin)基因转染治疗人卵巢癌裸鼠皮下移植瘤的作用及其相关机理。方法:使用人卵巢癌细胞株SKOV3建立人卵巢癌裸鼠皮下移植瘤模型。随机将荷瘤裸鼠分为基因治疗组、空质粒组和空白对照组,分别于瘤体注射Angiosta-tin/PCDNA3、空质粒PCDNA3和无菌生理盐水,质粒用脂质体DOTAP介导转染细胞。观察各组动物的肿瘤生长曲线,检测各组肿瘤Angiostatin、VEGF的表达和微血管密度(MVD),利用TUNEL染色法行原位细胞凋亡分析,计算细胞凋亡指数(AI)。结果:基因治疗组裸鼠的肿瘤生长在早期受到明显抑制,大约1周后生长速度有所加快,肿瘤组织中Angiostatin蛋白局部高表达,VEGF的表达高于空质粒组和空白对照组,AI(4.21±0.49)高于空质粒组和空白对照组,MVD(19.3±3.2)低于空质粒组。结论:Angiostatin基因可以通过抑制血管生成而在一定程度上抑制肿瘤生长,其作用的发挥是独立于VEGF的。

Experimental study of treating implanted human ovarian carcinoma of nude mice by angiostatin gene in vivo

Objective:To observe the effect of angiostatin gene on implanted human ovarian carcinoma of nude mouse.Methods:Tumors were established by injection of 3×106 tumor cells named SKOV3 into the back of nude mice.The mice were classified randomly into three groups,injected respectively with angiostatin plasmid,empty plamid PCDNA3 and normal sodium.Half of the nude mice were used to observe the tumor growth curve,the other half to obtain the tumor samples.The tumor samples were prepared in the 5th day after gene transfer for the examinations of the expression of angiostatin and VEGF with immunohistochemistry,of MVD in the tumor with immunohistochemistry,and of cell apoptosis with TUNEL staining.Results:In the angiostatin group,the tumors were suppressed significantly in the first 7 days after gene transfer,and then they growed fast again.Immunohistochemistry analysis normal sodium revealed overexpression of angiostatin in situ.In addition,the expression of VEGF in the angiostatin group was a little higher than that in the control group.AI in the angiostatin group(4.21±0.49) was much higher than that in the control group(P<0.01),concurrently MVD(19.3±3.2) was lower (P<0.05).Conclusion:Angiostatin gene therapy can inhibit the growth of ovarian cancer in a certain extent,and this process is independent of VEGF.