骨桥蛋白反义基因对胃癌浸润与转移影响的研究

Effect of anti-sense osteopontin on metastasis and infiltration of gastric carcinoma

目的:探讨骨桥蛋白反义基因对人类胃癌细胞增殖和浸润转移的影响。方法:1)PCR扩增获得人OPN基因,pcD-NA3.1(+)载体连接,酶切、测序。2)脂质体介导将pcDNA3.1-ANOPN基因转入ECA109,G418筛选,获得稳定表达AN-OPN基因的转染细胞SGC-7901-ANOPN、表达空载体的SGC-7901-vect细胞和空白对照SGC-7901。RT-PCR检测细胞OPNmRNA表达,体外观察对比各组间倍增时间、黏附、侵袭及迁移能力的差异。结果:成功构建了pcDNA3.1-ANOPN质粒,测序结果与GenBank中公布的序列同源性为100%。与SGC-7901-vect、SGC-7901相比,SGC-7901-ANOPN细胞OPNmRNA表达明显降低,倍增时间增加,黏附、侵袭及迁移能力明显降低。结论:ANOPN基因的稳定表达明显抑制胃癌细胞的恶性表型。

OBJECTIVE：To investigate the effect of anti-sense osteopontin（ANOPN）gene on the proliferation,infiltration and metastasis of gastric carcinoma cells.METHODS：An OPN gene recombinant expression vector plasmid was constructed by RT-PCR from human umbilical vein endothelial cell gene and cloned into a mammalian expression vector pcDNA3.1（＋）.PcDNA3.1-ANOPN was introduced by Lipofectin to ECA109.Positive cell clones SGC-7901-ANOPN,vector transfected cells SGC-7901-vect and blank cell SGC-7901 were used as three groups.RT-PCR was used to investigate the expressions of OPN mRNA.The metastasis characteristics of the cells were studied by the Transwell method.RESULTS：The vector was constructed successfully,and the sequencing result was 100% identical with that reported in GenBank.Compared with vector-transfected cells SGC-7901-vect cells and SGC-7901 cells,the growth rate of SGC-7901-ANOPN cells was significantly slowed,the expression rate was decreased,their doubling time was increased,and the number of SGC-7901-ANOPN cells in mucosa was decreased.CONCLUSION：The stable expression of ANOPN gene can significantly suppress the malignant phenotype of SGC-7901 cells.