上调 Notch 配体 DLL4 对人胃癌细胞生物学行为的影响。
Influence of up-regulation of Notch ligand DLL4 on biological behaviors of human gastric cancer cells.
机构信息
Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, Zhejiang Province, China.
出版信息
World J Gastroenterol. 2013 Jul 28;19(28):4486-94. doi: 10.3748/wjg.v19.i28.4486.
AIM
To investigate the potential roles of Delta-like ligand 4 (DLL4) on the biological behavior of gastric cancer cells and its molecular mechanisms.
METHODS
A recombinant eukaryotic expression vector containing human DLL4 gene was constructed and transfected into the human gastric cancer cell line SGC7901. Clones with up-regulated DLL4 were selected and amplified. The effect of DLL4 up-regulation on gastric cancer cell growth was assessed using cell growth assay. The migration and invasion were assessed using a transwell migration assay and matrigel invasion assay. Matrix metalloproteinases were detected using the zymogram technique. Cells were implanted subcutaneously into male BALB/c nu/nu mice. Tumor volumes were then calculated and compared. DLL4 staining in the implanted tumor was performed using immunohistochemistry technique.
RESULTS
Growth curves over a six-day time course showed significantly promoted cell proliferation of SGC7901 cells with up-regulated DLL4. DLL4 up-regulation in SGC7901 cells promoted the migration (205.4 ± 15.2 vs 22.3 ± 12.1, P < 0.05) and invasion (68.8 ± 5.3 vs 18.2 ± 6.0, P < 0.05) in vitro and tumorigenicity in vivo (2640.5 ± 923.6 mm(3) vs 1115.1 ± 223.8 mm(3), P < 0.05). Furthermore, significantly increased mRNA level and increased secretion of matrix metalloproteinase-2 (MMP-2) proenzyme were observed in SGC7901 cells with up-regulated DLL4. However, increased MMP-9 mRNA level but decreased extracellular MMP-9 proenzyme level was observed.
CONCLUSION
Our observations indicated a mechanism by which activation of DLL4-mediated Notch signaling promotes the expression and secretion of MMP-2 proenzyme and influences the progress of gastric cancer.
目的
探讨 Delta 样配体 4（DLL4）在胃癌细胞生物学行为中的潜在作用及其分子机制。
方法
构建了含有人 DLL4 基因的重组真核表达载体，并转染人胃癌细胞系 SGC7901。选择并扩增上调 DLL4 的克隆。通过细胞生长试验评估 DLL4 上调对胃癌细胞生长的影响。通过 Transwell 迁移试验和 Matrigel 侵袭试验评估迁移和侵袭。使用酶谱技术检测基质金属蛋白酶。将细胞皮下植入雄性 BALB/c nu/nu 小鼠。然后计算并比较肿瘤体积。使用免疫组织化学技术检测植入肿瘤中的 DLL4 染色。
结果
六天生长曲线显示，上调 DLL4 的 SGC7901 细胞增殖明显加快。SGC7901 细胞中 DLL4 的上调促进了体外迁移（205.4 ± 15.2 比 22.3 ± 12.1，P < 0.05）和侵袭（68.8 ± 5.3 比 18.2 ± 6.0，P < 0.05），以及体内致瘤性（2640.5 ± 923.6 mm3 比 1115.1 ± 223.8 mm3，P < 0.05）。此外，上调 DLL4 的 SGC7901 细胞中 MMP-2 前酶的 mRNA 水平和分泌量显著增加，而 MMP-9 的 mRNA 水平增加，细胞外 MMP-9 前酶水平降低。
结论
我们的观察结果表明，DLL4 介导的 Notch 信号激活促进了 MMP-2 前酶的表达和分泌，并影响了胃癌的进展。
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