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目的:研究血管抑制素基因体外对结肠癌细胞增殖的抑制及体内对肿瘤血管生成的抑制,探讨血管抑制素基因对结肠癌的抑制作用。方法:构建重组质粒pcDNA3.1(+)／angio,用脂质体转染法将重组质粒、空白载体导入结肠癌细胞Colo205,培养细胞观察细胞生长,绘制细胞生长曲线,计算增殖抑制率,然后将pcDNA3.1(+)／angio、pcDNA3.1(+)／Colo205、Colo205分别接种至裸鼠右侧颈部皮下,观察肿瘤的生长,测量肿瘤体积,计算抑瘤率,免疫组化检测肿瘤组织中微血管密度(MVD)、血管内皮生长因子(VEGF)的表达。结果:体外pcDNA3.1(+)／angio组细胞的生长速度略慢于pcDNA3.1(+)／Colo205、Colo205组,但差异无统计学意义(P>0.05);体内pcDNA3.1(+)／angio组细胞的致力显著降低,瘤体增长缓慢,抑瘤率较高(P<0.01),瘤体组织中MVD及VEGF的表达较低(P<0.05);pcDNA3.1(+)／Colo205、Colo205两组之间的差异无统计学意义(P>0.05)。结论:在体外血管抑制素不直接影响结肠癌细胞Colo205的生物学特性,但在体内可强烈抑制肿瘤的生长,其机制可能是通过降低血管生成因子而抑制肿瘤的新生血管生成,发挥抑制肿瘤作用。
OBJECTIVE: To study the inhibitory effect of angiostatin gene on the proliferation of colon cancer cells in vitro and the inhibition of tumor angiogenesis in vivo, and to investigate the inhibitory effect of angiostatin gene on colon cancer. Methods: The recombinant plasmid pcDNA3.1 (+) / angio was constructed. The recombinant plasmid and vector were transfected into Colo205 colon cancer cells by lipofection. Cell growth was observed and the cell growth curve was drawn. The growth of tumor was observed by inoculating pcDNA3.1 (+) / angio, pcDNA3.1 (+) / Colo205 and Colo205 subcutaneously into the right neck of nude mice respectively. The tumor volume was measured and the tumor inhibition rate was calculated. Immunohistochemical detection of tumor Tissue microvessel density (MVD), vascular endothelial growth factor (VEGF) expression. Results: The growth rate of pcDNA3.1 (+) / angio group in vitro was slightly slower than that of pcDNA3.1 (+) / Colo205 and Colo205 group, but the difference was not statistically significant (P> 0.05) (P <0.01). The expression of MVD and VEGF in tumor tissue was lower (P <0.05). The expression of pcDNA3.1 (+) / Colo205 , Colo205 difference between the two groups was not statistically significant (P> 0.05). Conclusion: In vitro angiostatin does not directly affect the biological characteristics of colon cancer cells Colo205, but in vivo can strongly inhibit tumor growth, the mechanism may be by reducing angiogenic factors and inhibit tumor angiogenesis, play a tumor suppressor role .