缺氧微环境对人肺癌高转移细胞株95D失巢凋亡的研究

The Effect of Hypoxia on Anoikis in High-metastatic Human Lung Cancer Cell Line 95D

目的研究缺氧微环境对人巨细胞肺癌高转移细胞系95D失巢凋亡的影响。方法对95D悬浮培养,建立人肺癌高转移细胞株95D失巢凋亡抵抗细胞模型,实验组进行缺氧培养,对照组正常有氧培养,MTT检测缺氧微环境对于失巢凋亡抵抗95D细胞系生长的影响,流式细胞仪检测2组细胞的凋亡情况,Transwell实验检测2组细胞侵袭能力,Western blot检测缺氧对于失巢凋亡抵抗95D细胞中HIF-1α、VEGF和MMP-2蛋白表达的影响。结果缺氧微环境下失巢凋亡抵抗细胞组生长率为52.9%;凋亡率(40.4%)明显高于非缺氧培养组(21.7%),差异有统计学意义(P<0.05);侵袭能力降低,穿过膜细胞数(40.1±6.7)明显低于对照组(12.5±7.9),差异有统计学意义(P<0.05);缺氧微环境下失巢凋亡抵抗95D细胞中HIF-1α表达上调,侵袭转移相关蛋白VEGF和MMP-2表达下调。结论在人肺癌高转移细胞株95D失巢凋亡过程中,缺氧微环境能从一定程度上抑制失巢凋亡抵抗细胞的生长,缺氧微环境能降低失巢凋亡抵抗细胞的生存能力和转移能力,但这可能是缺氧后早期表现。

Objective To investigate the effect of hypoxia micro-environment on anoikis in human high-metastatic cell line 95D of giant cell carci-noma of lung. Methods Suspension technology was used to culture 95D to establish the model of anoikis-resistant 95D cells. Hypoxic culture was conducted in the experimental group,and regular culture was conducted in the control group. The effect of hypoxia on proliferation of anoikis-resis-tant 95D was investigated by MTT and the apoptosis in the two groups were detected by flow cytometer. The invasive ability of the cells was assessed by Transwell test. The effect of hypoxia on the expression of HIF-1α,VEGF and MMP-2 in anoikis-resistant 95D was detected by Western blot. Re-sults The growth rate of the anoikis-resistant 95D cells treated with hypoxia was 52.9%,and the apoptosis rate of these cells was higher than that in the non-hypoxic group（40.4%vs 21.7%,P〈0.05）. The treatment of hypoxia down-regulated the invasive ability,the number of migration cells un-der hypoxia was higher than that in the control group,with statistical significance（40.1±6.7 vs 12.5±7.9,P〈0.05）. The up-regulation of HIF-1α, the down-regulation of VEGF and MMP-2 were observed in the group of hypoxia. Conclusion During anoikis of human high-metastatic lung can-cer cell line 95D,hypoxia inhibited the survival ability and the metastasis ability of anoikis-resistant cells,which,however,might be the early mani-festation of hypoxia.