慢病毒介导shRNA沉默survivin基因对鸡胚绒毛尿囊膜子宫内膜异位症模型的影响

Effect of lentiviral vector-mediated short hairpin RNA targeting survivin against endometriosis-like lesions in chick embryo chorioallantoic membrane

目的研究慢病毒介导生存素基因特异性短发夹状RNA(shRNA)对子宫内膜异位症异位内膜生长及新生血管形成的影响。方法建立鸡胚绒毛尿囊膜(CAM)子宫内膜异位症模型,随机分成单纯接种组、shRNA慢病毒组、空载慢病毒组和空白载体组,每组30只。通过Image-proplus软件测量新生血管面积与鸡胚尿囊膜面积比(VA/CAM)评价沉默survivin基因对CAM血管生成的影响,TUNEL检测异位内膜细胞凋亡,组织病理学观察异位内膜病灶生长行为。结果 shRNA慢病毒处理组种植内膜存活率最低,该组VA/CAM明显低于单纯接种组、空载慢病毒组和空白载体组(P<0.001),后3组间比较无显著性差异(P>0.05)。shRNA慢病毒处理组种植内膜细胞凋亡率明显高于其他3组(P<0.05),另3组间比较无显著性差异(P>0.05)。shRNA慢病毒处理组病灶在光镜下可见腺体结构不完整,间质伴有不同程度的坏死。结论慢病毒介导survivin基因特异性shRNA能够显著抑制人子宫内膜异位症种植CAM模型的新生血管形成,抑制子宫内膜在CAM上的种植生长。

Objective To investigate the inhibitory effect of lentiviral vector-mediated short hairpin RNA targeting survivin （LV-survivin shRNA） on angiogenesis and growth of endometriosis-like lesions in chick en embryo chorioallantoic membrane. Methods Eutopic endometrium from women with endometriosis was transplanted onto the non-vascular region of （CAM）, where LV-survivin shRNA was delivered subsequently. The angiogenesis and the growth of endometriosis-like lesions in the CAM model were evaluated. Results The angiogenesis and formation of endometriosis-like lesions were significantly suppressed in the CAM model by treatment with LV-survivin shRNA in comparison with those in the untreated CAM models and models treated with empty LV or DMEM （P〈0.001）. LV-survivin shRNA also caused a significantly higher cell apoptotic rate in the endometriosis-like lesions than the other treatments （P〈0.001） and induced necrosis in the lesions. Conclusion LV-survivin shRNA can effectively inhibit angiogenesis induced by the eutopic endometrium and markedly suppress the formation of endometriosis-like lesions in the CAM model.