目的 观察NDRG2基因对人前列腺癌细胞株PC-3M侵袭转移能力的影响.方法 以携带人NDRG2基因的腺病毒感染体外培养的PC-3M细胞株,采用Western blot及明胶酶谱实验检测NDRG2、MMP-2、MMP-9蛋白表达情况及相关酶活性的变化.平板克隆实验、细胞生长实验检测NDRG2对PC-3M增殖能力的影响.Transwell实验检测NDRG2对PC-3M细胞体外侵袭能力的影响.结果 Ad-NDRG2感染后,PC-3M细胞中NDRG2表达明显增加,而MMP-2和MMP-9表达水平及活性均降低.噻唑蓝(MTT)比色法(抑制率24 h为16.2%、48 h为24.4%、72 h为43.7%)及平板克隆实验(3组分别为56.3%、55.2%和36.7%)显示NDRG2对PC-3M细胞的生长有明显抑制作用.Transwell显示对照组及Ad-LacZ组穿入下室面的细胞数明显多于Ad-NDRG2组(3组分别为93.0、94.8和50.4).结论 NDRG2对人前列腺癌PC-3M细胞株侵袭能力有明显抑制作用,提示NDRG2可能在前列腺癌的侵袭及转移中发挥重要作用.
Objective To investigate the effect of NDRG2 gene expression on the cell migration and invasion of human prostate cancer cell line PC-3M.Methods Recombinant adenovirus vectors carrying human NDRG2 gene (Ad-NDRG2) were infected into prostate cancer cell line PC-3M.The protein expression and enzymatic activities of NDRG2,MMP-2 and MMP-9 were determined by Western blotting and gelatin zymography respectively.Methylthiazol tetrazolium (MTT) assay and plate colony formation were used to determine the effect of proliferation of PC-3M cells.Invasion of PC-3M cells was measured by Transwell chamber assay.Results After infection by Ad-NDRG2,it had been verified that the protein expression of NDRG2 in PC-3M cells was obviously increased,and the expression and enzymatic activities of MMP-2 and MMP-9 were reduced.The MTT assay (inhibition ratio: 24 h,16.2%,48 h,24.4%,and 72 h,43.7% respectively) and plate colony formation (56.3%,55.2% and 36.7% in control group,Ad-LacZ group and Ad-NDGR2 group respectively) revealed that NDRG2 could significantly inhibit the growth and proliferation of PC-3M cells.The number of PC-3M cells that invaded the lower chamber in the Ad-NDRG2 group was significantly decreased as compared with the control group and the Ad-LacZ group (93.0,94.8 and 50.4 respectively).Conclusion NDRG2 gene can significantly inhibit invasion of the PC-3M cells,which may paly an important role in metastasis of prostate cancer.