论文部分内容阅读
目的探讨脂质体介导的内皮抑素真核表达载体对裸鼠移植性卵巢癌的抑制作用。方法利用已构建的内皮抑素真核表达载体pVAX1sEn,在脂质体介导下转染卵巢癌细胞系3AO细胞,RTPCR技术检测3AO细胞中内皮抑素mRNA的表达,酶联免疫吸附实验(ELISA)法检测3AO细胞上清液中内皮抑素的水平,四甲基偶氮唑蓝(MTT)法检测3AO细胞上清液中的内皮抑素对脐静脉内皮细胞系ECV204细胞的抑制作用。将荷卵巢癌移植瘤裸鼠分为3组,pVAX1sEn治疗组、pVAX1对照组和生理盐水对照组,分别观察3组卵巢癌的生长情况。结果RTPCR技术检测结果显示,在610bp处有一内皮抑素mRNA的特异性条带;ELISA法检测结果显示,pVAX1sEn转染3AO细胞后其上清液中内皮抑素水平为(201±8)ng/ml;MTT法检测结果显示,pVAX1sEn转染3AO细胞后其上清液中的内皮抑素可有效抑制ECV204细胞的生长,最高抑制率为42%。pVAX1sEn治疗组肿瘤体积为(0.85±0.18)cm3,显著小于生理盐水对照组的(1.90±0.28)cm3和pVAX1对照组的(1.78±0.32)cm3(P<0.05)。肿瘤组织HE染色显示,pVAX1sEn治疗组肿瘤细胞坏死明显,而生理盐水对照组及pVAX1对照组肿瘤细胞生长旺盛。结论脂质体介导的pVAX1sEn瘤内注射可有效抑制卵巢癌移植瘤的生长。
Objective To investigate the inhibitory effect of liposome-mediated endostatin eukaryotic expression vector on transplanted ovarian cancer in nude mice. Methods The constructed endostatin eukaryotic expression vector pVAX1sEn was transfected into 3AO cells by lipofectamine. The expression of endostatin mRNA in 3AO cells was detected by RTPCR. The expression of endostatin mRNA was detected by enzyme - linked immunosorbent assay (ELISA) ) Method was used to detect the level of endostatin in 3AO cell supernatant. The inhibitory effect of endostatin in 3AO cell supernatant on human umbilical vein endothelial cell line ECV204 was detected by MTT assay. Nude mice bearing ovarian cancer xenografts were divided into 3 groups: pVAX1sEn treatment group, pVAX1 control group and saline control group. The growth of 3 groups of ovarian cancer were observed. Results The results of RTPCR showed that there was a specific band of endostatin mRNA at 610bp. The results of ELISA showed that the endostatin level in the supernatant of the transfected 3AO cells was (201 ± 8) ng / ml. The results of MTT assay showed that endostatin in supernatant of pVAX1sEn transfected 3AO cells could effectively inhibit the growth of ECV204 cells with the highest inhibition rate of 42%. The tumor volume in the pVAX1sEn treated group was (0.85 ± 0.18) cm3, which was significantly lower than that in the saline control group (1.90 ± 0.28) cm3 and (1.78 ± 0.32) cm3 in the pVAX1 control group (P <0.05). Tumor tissue HE staining showed that pVAX1sEn treatment group significantly tumor cell necrosis, while saline control group and pVAX1 control group tumor cell growth. Conclusion Liposome-mediated intratumoral injection of pVAX1sEn can effectively inhibit the growth of ovarian cancer xenografts.