摘要
AIM： To investigate the effect of angiopoietin-1 （Ang-1） on biological behaviors in vitro and tumorigenesis and angiogenesis in vitro of human gastric cancer cells. METHODS： Human full-length Ang-1 gene was cloned from human placental tissues by RT-PCR method. Recombinant human Ang-1 antisense eukaryotic expression vector was constructed by directional cloning, and transfected by lipofectin method into human gastric cancer line SGC7901 with high Ang-1 expression level. Inhibition efficiency was confirmed by semi- quantitive PCR and Western blot method. Cell growth curve and cell cycle were observed with MTT assays and flow cytometry, respectively. Nude mice tumorigenicity test was employed to compare in vitro tumorigenesis of cells with Ang-1 suppression. Microvessel density （MVD） of implanted tumor tissues was analyzed by immunohistochemistry for factor VIII staining. RESULTS： Full-length Ang-1 gene was successfully cloned and stable transfectants were established, namely 7Ang1- for antisense, and 7901P for empty vector transfected. 7Ang1- cells showed down-regulated Ang-1 expression, while its in vitro proliferation and cell cycle distribution were not significantly changed. In contrast, xenograft of 7Ang1- cells in nude mice had lower volume and weight than those of 7901P after 30 days＇ implantation （P〈 0.01, 293.00：1±95.54 mg vs. 624.00±77.78 mg） accompanied with less vessel formation with MVD 6.00±1.73 compared to 7901P group 8.44±1.33 （P〈 0.01）. CONCLUSION： Ang-1 may play an important role in tumorigenesis and angiogenesis of gastric cancer, and targeting its expression may be beneficial for the therapy of gastric cancer.
瞄准：在生物行为在试管内和人的胃的癌症房间的血管生成在试管内上调查 angiopoietin-1 (Ang-1 ) 的效果。方法：人的全身的 Ang-1 基因被 RT-PCR 方法从人的胎盘的纸巾克隆。Recombinant 人的 Ang-1 反察觉到真核细胞的表示向量被与高 Ang-1 表示水平的进人的胃的癌症线 SGC7901 的由 lipofectin 方法的方向性的克隆，和 transfected 构造。抑制效率被半证实 -- 量的 PCR 和西方的污点方法。房间生长曲线和房间周期分别地与 MTT 试金和流动血细胞计数被观察。裸体老鼠 tumorigenicity 测试被采用把房间的在试管内肿瘤发生与 Ang-1 抑制作比较。植入的肿瘤纸巾的 Microvessel 密度(MVD ) 被免疫组织化学为第八因子染色分析。结果：全身的 Ang-1 基因成功地被克隆，稳定的 transfectants 被建立，也就是为为空向量 transfected 的 antisenseiand 7901P 的 7Ang1- 。7Ang1- 房间显示出下面调整的 Ang-1 表示，当它的在试管内增长和房间周期分发显著地没被改变时。相反，在裸体老鼠的 7Ang1- 房间的异种皮移植在 30 白天与 7901P 相比与 MVD 6.00+/-1.73 伴有更少的容器形成的培植(P<0.01， 293.00+/-95.54 mg 对 624.00+/-77.78 mg ) 以后 7901P 比那些让更低的体积和重量组织 8.44+/-1.33 (P<0.01 ) 。结论：Ang-1 可以在胃的癌症的肿瘤发生和血管生成起一个重要作用，并且指向它的表示可能为胃的癌症的治疗有益。
基金
Supported by the National Natural Science Foundation of China, No. 30130260, 30225039
