MTDH 基因下调抑制人乳腺癌 MDA-MB-453细胞增殖同黏附和迁移的研究

Down-regulation in MTDH Inhibition of Proliferation,Adhesion and Migration of Human Breast Cancer MDA-MB-453 Cells

目的:探讨MTDH基因对乳腺癌MDA-MB-453细胞增殖、黏附和迁移能力的影响。方法:采用RNA干扰技术,下调乳腺癌MDA-MB-453细胞MTDH基因的表达.RT-PCR和免疫细胞化学技术检测MTDH下调效果.通过细胞增殖实验、黏附实验和迁移实验分别检测MTDH基因表达下调后细胞增殖、黏附和迁移能力的变化。结果:MTDH-siRNA的转染效率达到90%,转染48 11后实验组细胞MTDH的mRNA和蛋白质表达较对照组分别降低了45.8<%和47.5%MTDH表达下调后,细胞增殖受到明显抑制,48 h和72 h抑制率分别为41.5%和49.0%;细胞黏附力下降,30min和60 imn黏附率较对照组分别降低42.0%和49.7%;细胞迁移能力显著降低,迁移率下降333%。结论:下调MTDH基因表达可明显抑制乳腺癌MDA-MB-453细胞的增殖、黏附和迁移能力,提示其在乳腺癌细胞的恶性生物学行为中发挥重要作用。

Objective： The current work aims to investigate the effects of metadherin （ MTDH ） down-regulation on cell proliferation, adhesion, and migration of human breast cancer MDA-MB-453 cells. Methods： RNA interference was employed to reduce MTDH expression in human breast cancer MDA-MB-453 cells. Reverse transcription polymerase chain reaction （ RT-PCR ） and immunocytochemistry were applied to ascertain the down-regulation of MTDH. 3-（ 4, 5-Dimethylthiazol-2-yl ） -2, 5-diphenyltetrazolium bromide assay, cell adhesion, and migration assay were performed to respectively assess the changes in cell proliferation, adhesion, and migration. Results： The transfection efficiency of MTDH-siRNA reached 90 %. Forty-eight hours after transfection, the expression of MTDH mRNA and protein was reduced by 45.8 % and 47.5 %, respectively, compared with the control group. The knockdown of MTDH inhibited cell proliferation, and the inhibition ratio was 41.5 % and 49.0 % at 24 h and 48 h, respectively. Cell adhesion was also inhibited. The adhesion rate at 30 min and 60 min was decreased by 42.0 % and 49.7 %, respectively, compared with the control. Moreover, MTDH down-regulation resulted in a decreased migration rate of 33.3 %. Conclusion： The reduced MTDH expression in MDA-MB-453 cells can inhibit proliferation, adhesion, and migration, which suggests that MTDH plays an important role in the malignant biological behaviors of breast cancer cells.