Gene expression alterations in chronic hypoxic MCF7 breast cancer cell line

• Breast cancer cells (MCF7) were exposed to 8-hour hypoxic episodes three times a week for a total of 38 episodes.
• HNF4A has been up-regulated by 2 folds after 19 hypoxic shots and further up-regulated by 6.43 folds after 38 hypoxic shots.
• The IC50 of doxorubicin in MCF7 cells has increased in a trend proportional to the number of hypoxic episodes.
• The resistance toward doxorubicin totally rebounded after incubation under normoxia for 3 weeks.

Hypoxia plays a significant role in tumor progression and aggressiveness and implicated in resistance to radiotherapy and chemotherapy. This study aims to characterize the changes in gene expression associated with chronic hypoxia in MCF7 breast cancer cell line and identify a possible biomarker for hypoxia in breast cancer. Breast cancer cells (MCF7) were exposed to 8-hour hypoxic episodes (< 1% oxygen) three times a week for a total of 38 episodes. Gene expression changes were profiled using RT- PCR array after 19 and 38 episodes of hypoxia and compared to normoxic cells. Chemoresistance of hypoxic cells toward doxorubicin was assessed using MTT cell proliferation assay. Marked gene expression changes were indentified after 19 and 38 episodes of hypoxia. Only few changes were common in both stages with most genes rebounding at the level of 38 episodes. A notable gene (HNF4A) has been up-regulated by 2 folds after 19 hypoxic shots and further up-regulated by 6.43 folds after 38 hypoxic shots. The half maximal inhibitory concentration (IC50) of doxorubicin in MCF7 cells has increased in a trend proportional to the number of hypoxic episodes then totally rebounded after incubation under normoxia for 3 weeks. This study provides evidence that exposing cells to prolonged periods of hypoxia (weeks) results in different expression changes than those induced by short-term hypoxia (less than 72 h).