Effects Of NDRG1 On Proliferation And Invasiveness Of Human Colorectal Cancer Cell Line HT-29 In Vitro

Purpose:To investigate the effects of NDRG1 on proliferation and invasiveness of human colorectal cancer cell line HT-29 in vitro.Methods:Lipofectamine^TM2000 was used to stably transfect recombinant plasmids of pEGFP-NDRG1-N3 into human colorectal cancer cell line HT-29. After being sieved by G418,NDRG1 overexpression in the positive clones was determined by RT-PCR and Western blot analysis. Proliferation of HT-29 was measured by drawing Growth curve,counting growth rate of cell colony on the soft agar and fluorescence-activated cell sorting(FACS).The ability of migration(without Matrigel)and invasiveness(with Matrigel)of HT-29 was measured by counting the number of cells transferring through the polycarbonate membrane with 8μm pore in 24-transwell chambers.Results:Stable cell clones with marked overexpression of NDRG1 mRNA and protein were established.Proliferation,as measured by Growth curve and growth rate of cell colony on the soft agar,declined significantly in an HT-29 clone overexpressing NDRG1(P＜0.01),with an significant increase in cells at the G₀/G₁ phase and a significant decrease in cells at the S phase of cell cycle(P＜0.01).The number of cells transferring through the polycarbonate membrane with 8μm pore in 24-transwell chambers decreased significantly in an HT-29 clone overexpressing NDRG1(P＜0.01),in spite of with Matrigel(measuring the he ability of invasiveness)or without Matrigel(measuring the ability of migration).Conclusions:NDRG1 overexpression reduces HT-29 cells proliferation,migration and invasiveness potential in vitro.