转染NDRG1基因对宫颈癌细胞COX-2、VEGF表达及增殖的影响

目的： 通过研究人类N-myc下游调节基因1(N-myc downstream regulated gene 1,NDRG1)对人宫颈癌SiHa细胞环氧合酶-2(cyclooxygenase-2，COX-2)、血管内皮生长因子(vascular endothelial growth factor，VEGF)表达及细胞增殖的影响，深入探讨NDRG1抑制宫颈癌侵袭转移的作用机制。方法： 经脂质体介导将含有NDRG1真核表达质粒转染人宫颈癌SiHa细胞株，采用G418筛选阳性细胞克隆及RT-PCR、蛋白质印迹鉴定；蛋白质印迹法检测阳性克隆细胞COX-2及VEGF表达的改变；噻唑盐（MTT）比色法检测阳性细胞克隆的增殖活性。结果： 成功建立高表达NDRG1的阳性SiHa细胞克隆（N-12），并证实其COX-2和VEGF蛋白表达及细胞增殖活性均显著降低。结论： NDRG1可能通过下调宫颈癌细胞COX-2、VEGF的表达及抑制细胞增殖而起到抑制宫颈癌进展的作用。

Effect of gene NDRG1 transfection on the expression of COX-2 and VEGF and proliferation in human cervical cancer cells

Objective: To investigate the effect of N-myc downstream regulated gene 1(NDRG1) on the proliferation in human cervical cancer cell line SiHa, with a focus on the expression of cyclooxygenase-2(COX-2) and vascular endothelial growth factor(VEGF).  Methods pcDNA3.0-NDRG1 and pcDNA3.0 were transfected into SiHa cells through lipofectamine and positive clones were screened by G418. RT-PCR and Western blot were employed to identify mRNA and protein expression of NDRG1 in SiHa cells, respectively. The expression of COX-2 and VEGF in positive clones was detected by Western blot. Cell viability was assessed by MTT method. Results Positive clone N-12 with NDRG1 over expression was successfully established. Compared with non-transfected control group, the expression of COX-2 and VEGF was decreased significantly, and the proliferation level of N-12 was also decreased significantly. Conclusion It is possible that NDRG1 could inhibit the development of cervical cancer by downregulating the expression of COX-2 and VEGF and the proliferation in SiHa cells.