Nie Ying-Ming, Dai Bi-Tao
Department of Hematology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2009 Jul;11(7):566-70.
To study the potential effect of hypoxia on invasion and metastasis of leukemia cell line K562.
K562 cells were cultured with the conventional method in vitro and treated with 1%, 3% and 5% oxygen for 24 hrs. The normoxic cultured K562 cells were used as the control group. Cell adhesion assay, cell migration assay and cell invasion assay were used to detect the adhesion, migration and invasion abilities of K562 cells. RT-PCR was used to measure the mRNA expression of HIF-1alpha, VEGF, MMP-2 and MMP-9. The protein level of HIF-1alpha was measured by Western blot.
Compared with the control group, the 3% and 5% oxygen treatment groups significantly increased the adhesion, migration and invasion abilities of K562 cells (p<0.05 or <0.01), and up-regulated the protein level of HIF-1alpha and the mRNA levels of HIF-1alpha,VEGF, MMP-9 and MMP-2 (p<0.05 or 0.01). However, there were no significant differences in the above indexes between the 1% oxygen treatment and the control groups.
Moderate hypoxia can enhance the abilities of invasion and metastasis of K562 cells, probably by an up-regulation of HIF-1alpha level and VEGF, MMP-2 and MMP-9 mRNA expression.
研究缺氧对白血病细胞系K562侵袭和转移的潜在影响。
采用常规方法体外培养K562细胞，分别用1%、3%和5%的氧气处理24小时。将常氧培养的K562细胞作为对照组。采用细胞黏附实验、细胞迁移实验和细胞侵袭实验检测K562细胞的黏附、迁移和侵袭能力。采用RT-PCR检测HIF-1α、VEGF、MMP-2和MMP-9的mRNA表达。采用蛋白质免疫印迹法检测HIF-1α的蛋白水平。
与对照组相比，3%和5%氧气处理组显著提高了K562细胞的黏附、迁移和侵袭能力（p<0.05或<0.01），并上调了HIF-1α的蛋白水平以及HIF-1α、VEGF、MMP-9和MMP-2的mRNA水平（p<0.05或0.01）。然而，1%氧气处理组与对照组上述指标无显著差异。
中度缺氧可能通过上调HIF-1α水平以及VEGF、MMP-2和MMP-9的mRNA表达来增强K562细胞的侵袭和转移能力。
