【摘要】 目的　了解甲状腺滤泡状癌细胞中导入内皮抑素 (Endostatin ,ES)基因对体内外生长的影响 ,并探讨其作用机制。方法　用逆转录病毒载体将ES基因导入FTC133 细胞中 ,采用RT PCR和ELISA检测导入基因在肿瘤细胞的表达。观察肿瘤在裸鼠模型中的生长状况。用抗CD31抗体对肿瘤切片进行免疫组织化学染色检测微血管 ,计算微血管密度 (microvesseldensity ,MVD)。检测血清中ES和VEGF浓度。 结果　顺利将ES基因导入到FTC13 3 中得到稳定表达ES的肿瘤细胞系FTC133 rvEndo。后者在体外增值速度与母系相似。而在裸鼠模型中 ,其肿瘤生长速度明显慢于母系。种植 33天后 ,FTC133 组肿瘤大小为 ( 30 0 2± 44 1)mm3,FTC13 3 rvEndo组为 ( 5 5 7± 15 2 )mm3,两组间差异有显著性 (P =0 0 0 0 2 )。免疫组化染色表明FTC133 rvEndo肿瘤中血管内皮细胞数明显减少 ,MVD为 34 86± 10 6 8;而对照组为 6 4 71± 17 0 5 ,差异有显著性 (P =0 0 0 2 0 )。带瘤鼠血清人VEGF浓度FTC133 rvEndo组为 ( 8 99± 0 6 5 )ng/L ,明显低于对照组 ( 2 9 34± 5 5 5 )ng/L( P =0 0 0 98)。结论　利用逆转录病毒载体可将ES基因转入甲状腺滤泡状癌细胞系中 ,该基因的导入通过抑制血管生成使肿瘤体内生长速度减慢。其机制可能与抑制肿瘤细胞?更多还原

【Abstract】 Objective To evaluate the effect of retroviral endostatin (ES) gene transfer to a follicular thyroid carcinoma cell line FTC 133 on its behavior in vitro and in vivo. Methods The murine ES gene was transferred into FTC 133 cells using a retroviral vector pFB-Neo-Endo to get a stable cell line FTC 133-rvEndo.RT-PCR and ELISA were employed to confirm the expression of ES by FTC 133-rvEndo.FTC 133-rvEndo and FTC 133 cells were then injected subcutaneously on nude mice and the tumor size was measured regularly. After the mice were sacrificed, the tumors were stained immunohistochemically (IHC) using anti-CD 31 antibody and microvessel density (MVD) was calculated. The serum concentrations of human VEGF and murine ES were detected then.Results The successful transferred cells (FTC 133-rvEndo) were selected. RT-PCR and ELISA confirmed the expression of ES. The ES concentration in conditioned medium of FTC 133-rvEndo was 29.3ng/mL while that of FTC 133 was undetectable.Compared to the parental cell line, FTC 133-rvEndo xenograft was inhibited significantly. The tumor volume of FTC 133-rvEndo group was (557±152)mm3,and that of FTC 133 group was (3002±441)mm3 on 33 days after tumor inoculation (P=0.0002). IHC staining shows that the endothelial cells in FTC 133-rvEndo were significantly less than which in FTC 133 tumor. MVD in FTC 133-rvEndo was significantly lower than that in FTC 133(P=0.0020). The serum concentration of human VEGF of bearing FTC 133-rvEndo tumor mice was (8.99±0.65)ng/L, which was lower than that of bearing FTC 133 tumor,(29.34±5.55)ng/L(P=0.0098).Conclusion Retroviral endostatin gene transfer inhibits FTC 133 xenograft through antiangiogenesis effect. The antiangiogenesis effect may be caused by inhibition of the secretion of VEGF by tumor cells.更多还原