Biomedical Engineering Center, Department of Anatomy, the Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, China.
Gynecol Oncol. 2011 Apr;121(1):174-80. doi: 10.1016/j.ygyno.2010.11.004. Epub 2010 Dec 15.
The study was to investigate the role of EFEMP1 in angiogenesis and growth of cervical carcinoma in vivo.
Effects of EFEMP1 on proliferation of Hela cells and HUVECs, invasion of Hela cells and migration of HUVECs, and adhesion of Hela cells to HUVECs were evaluated by MTT, Transwell chamber assay and adhesion assay, respectively. EFEMP1 overexpression in Hela cells was achieved by stable EFEMP1 gene transfection into Hela cells by Lipofectamin™ 2000 and the effectiveness of transfection was verified with western-blotting. The effect of EFEMP1 transfection upon the VEGF expression of Hela cells was detected with ELISA. The nude mouse models bearing cervical cancer were established with Hela cells transfected with EFEMP1 gene to observe the role of EFEMP1 in angiogenesis and growth of cervical cancer in vivo. VEGF expression and microvascular density of cervical cancer tissues were detected with immunohistochemistry and CD34 labeling respectively to elucidate the pathway by which EFEMP1 influences the growth of cervical cancer.
Proliferation and invasion of Hela cells were promoted by the EFEMP1 protein. The EFEMP1 gene transfection into Hela cells was successful and EFEMP1 gene obtained stable high expression in Hela cells. Compared to the control, the tumors with EFEMP1 overexpression showed a faster growth rate and had a higher level of VEGF expression and microvascular density. EFEMP1 gene transfection elevated the VEGF protein level in Hela cells and EFEMP1 protein facilitated the adhesion of Hela cells to HUVECs. However, no direct effect of EFEMP1 was observed on proliferation, migration and tube formation of HUVECs.
EFEMP1 promoted the angiogenesis and accelerated the growth of cervical carcinoma in vivo through a VEGF up-regulation pathway.
本研究旨在探讨 EFEMP1 在体内宫颈癌血管生成和生长中的作用。
通过 MTT、Transwell 室分析和黏附分析分别评估 EFEMP1 对 Hela 细胞和 HUVECs 增殖、Hela 细胞侵袭和 HUVECs 迁移以及 Hela 细胞与 HUVECs 黏附的影响。通过 Lipofectamin™ 2000 将 EFEMP1 基因稳定转染入 Hela 细胞中，实现 EFEMP1 在 Hela 细胞中的过表达，并通过 Western-blotting 验证转染的有效性。用 ELISA 检测 EFEMP1 转染对 Hela 细胞 VEGF 表达的影响。用转染 EFEMP1 基因的 Hela 细胞建立裸鼠宫颈癌模型，观察 EFEMP1 体内对宫颈癌血管生成和生长的作用。用免疫组织化学和 CD34 标记分别检测宫颈癌组织中 VEGF 表达和微血管密度，以阐明 EFEMP1 影响宫颈癌生长的途径。
EFEMP1 蛋白促进 Hela 细胞的增殖和侵袭。EFEMP1 基因成功转染入 Hela 细胞，EFEMP1 基因在 Hela 细胞中获得稳定的高表达。与对照组相比，过表达 EFEMP1 的肿瘤生长更快，VEGF 表达和微血管密度更高。EFEMP1 基因转染提高了 Hela 细胞中 VEGF 蛋白水平，EFEMP1 蛋白促进了 Hela 细胞与 HUVECs 的黏附。然而，EFEMP1 蛋白对 HUVECs 的增殖、迁移和管形成没有直接影响。
EFEMP1 通过上调 VEGF 途径促进体内宫颈癌的血管生成和加速生长。
