VDUP1对乳腺癌细胞MCF-7增殖及迁移的影响

Effects of VDUP1 on proliferation and migration of human breast cancer MCF-7 cells

目的:探讨过表达/沉默维生素D3上调蛋白1(vitamin D3 up-regulated protein 1,VDUP1)基因对人乳腺癌细胞系MCF-7增殖和迁移能力的影响及相关作用机制。方法:通过基因过表达/干扰技术上调/下调乳腺癌细胞系MCF-7中VDUP1基因的表达;实时荧光定量PCR检测细胞中VDUP1的mRNA表达水平;CCK-8法、Brd U实验和Transwell细胞迁移实验分别用于检测细胞增殖和迁移能力;Western blot检测细胞中Akt、p-Akt、GSK3β和p-GSK3β的蛋白水平。结果:基因过表达/干扰技术可上调/下调乳腺癌细胞系MCF-7中VDUP1基因的表达;过表达VDUP1基因后,MCF-7的细胞活力、DNA合成和细胞迁移能力显著降低(P<0.05),而沉默VDUP1基因后,MCF-7的细胞活力、DNA合成和细胞迁移能力则显著升高(P<0.05)。此外,过表达VDUP1基因可下调p-Akt和p-GSK3β的蛋白水平(P<0.05),而沉默VDUP1基因的结果则相反。结论:改变VDUP1基因表达水平可影响MCF-7细胞的增殖和迁移能力,其作用机制可能与Akt/GSK3β信号通路有关。

AIM ： To investigate the effect of vitamin D3 up-regulated protein 1 （ VDUP1） gene over-expres- sion/knockdown on the proliferation and migration of human breast cancer MCF-7 cells and its related mechanisms. METH-ODS ：Gene over-expression/interference techniques were used to up-regulate/down-regulate the expression of VDUP1 in the MCF-7 cells. The mRNA expression of VDUP1 was detected by qPCR. CCK-8 , BrdU and Transwell assays were used to measure the cell viability, proliferation and migration, respectively. The protein levels of Akt, p-Akt, GSK3p and p- GSK3p were determined by Western blot. RESULTS： The mRNA expression of VDUP1 was up-regulated after transfection with VDUP1 over-expression plasmid （ P 〈 0. 05 ） , and down-regulated after transfection with VDUP1 siRNA （P 〈 0. 05 ） . Over-expression of VDUP1 significantly inhibited MCF-7 cell proliferation and migration （ P 〈0. 05） , while knockdown of VDUP1 enhanced cell proliferation and migration （ P 〈 0. 05 ）. Furthermore, over-expression of VDUP1 up-regulated the protein levels of p-Akt and p-GSK3p （P 〈 0 .0 5 ） . Inverse results were obtained after knockdown of VDUP1. CONCLU-SION ：The viability and migration ability of MCF-7 cells are inhibited by over-expression of VDUP1 but enhanced by VDUP1 knockdown, which may be related with Akt/GSK3p pathway.