Abstract

Background

Carbonic anhydrase IX (CA IX) is a hypoxia-regulated transmembrane protein, implicated in cell adhesion, EMT, and tumour invasiveness, and is therefore strongly associated with cancer progression. It was previously found that shedding and release of CA IX ectodomain (ECD) into plasma of cancer patients can signify prognosis and is considered a marker for therapy success. In this work, we have investigated shedding of CA IX ECD in tumour cells undergoing apoptosis in response to cycloheximide and doxorubicin treatment.

Method

Using flow cytometry and cell sorting, we have correlated the extent of apoptosis with the presence of cell surface CA IX in CGL3 cell line characterized by both positive and negative CA IX subpopulations, and then determined level of shed ECD by ELISA.

Results

We have observed lower percentage of apoptotic cells in CA IX positive subpopulation after cycloheximide treatment. However, CA IX level of cycloheximide and doxorubicin-treated cells decreased during apoptotic death, suggesting an increased ECD shedding, that was not averted by inhibitors of apoptosis but decreased after metalloproteinase inhibition.

Conclusion

These findings supported the view that prevention of CA IX ECD shedding may contribute to better cancer cell survival.