目的观察缺氧微环境中缺氧诱导因子1α（HIF-1α）沉默的人绒毛膜癌细胞系JEG-3的侵袭、迁移能力。方法取对数生长期JEG-3细胞分为A、B、C、D组,其中A、B、C组置于体外缺氧微环境培养,D组不处理。当细胞融合到60%~70%时A、C组分别用HIF-1αsiRNA和无关序列转染,B、D组不做处理。采用Transwell细胞侵袭试验及划痕试验检测各组细胞侵袭能力和迁移能力。采用实时荧光定量PCR法和蛋白印迹法检测各组细胞HIF-1α、CXCR4的mRNA和蛋白表达。结果 A组细胞的侵袭和迁移能力明显低于其余3组（P均〈0.05）,B、C组细胞侵袭和迁移能力显著高于D组（P均〈0.05）。A组细胞中HIF-1α、CXCR4的mRNA和蛋白表达均低于其余3组（P均〈0.05）,B、C组细胞HIF-1α蛋白、CXCR4 mRNA和蛋白表达均高于D组（P均〈0.05）。结果缺氧微环境中HIF-1α沉默的JEG-3细胞侵袭、迁移能力下降,其机制可能与HIF-1α下调细胞CXCR4表达有关。
Objective To investigate the effects of hypoxia inducible factor 1α（ HIF-1α） on the invasion and migration of human choriocarcinoma JEG-3 cells under hypoxia. Methods JEG-3 cells in the logarithmic phase were divided into groups A,B,C,and D; and cells in the groups A,B,and C were cultured under hypoxia in vitro,but the group D was not treated. When JEG-3 cells were fused to 60% to 70%,cells in the groups A and C were transfected with HIF-1αsiRNA and NC siRNA,respectively,and we did not do the treatment on the cells in the groups B and D. Real-time PCR and Western blotting were used to detect the mRNA and protein expression levels of HIF-1α and CXCR4 in each group,respectively. The invasion and migration abilities of JEG-3 cells were separately analyzed by Transwell invasion assay and Scratch test. Results The invasion and migration abilities of JEG-3 cells in the group A were significantly lower than those in the other 3 groups（ all P〈0. 05）,the invasion and migration abilities of JEG-3 cells in the groups B and C were higher than those in the group D（ all P〈0. 05）. The mRNA and protein expression levels of HIF-1α and CXCR4 in the group A were significantly lower than those in the other 3 groups（ all P〈0. 05）,while the mRNA and protein expression levels of HIF-1α and CXCR4 in the groups B and C were significantly higher than those in the group D（ all P〈0. 05）.Conclusion Silencing HIF-1α can inhibit the invasion and migration abilities of of JEG-3 cells under hypoxia,and its molecular mechanism may be related to the down-regulation of CXCR4.
Shandong Medical Journal
hypoxia-inducible factor 1α
chemokine receptor 4