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.