Cancer stem cell isolation and characterization from lung cancer cells

Abstract

Lung cancer is one of the most common cancers in the world and a leading cause of death. Cancer stem cells (CSCs) are the cells responsible for tumor initiation. CSCs features include self-renewal capacity, differentiation, high invasion-migration, and resistance to chemotherapy. CSCs are characterized by specific surface markers. CD133 is widely used as a marker of CSCs in many cancers, including lung cancer. CD133-positive cells are an important marker of lung cancer stem cells because they show characteristics of lung cancer CSCs. CSCs are less sensitive and/or resistant to conventional treatments, remaining viable and regenerating tumors after treatment. Treatments that can target CSCs will be able to destroy CSCs more effectively, leading to the disappearance of tumors. Currently, attempts are being made to develop therapeutics that can effectively target CSCs. These studies concern the isolation and characterization of CSCs. In this study, CD133-positive CSCs and CD133-negative cells were isolated from H460 cells, a large cell lung cancer model, by MACS method. CSCs properties were investigated by tumorSphere formation assay (2% agar plate), hanging drop assay and qPCR method. CD133 positivity of H460 cells was determined as 0.3%. It has been observed that CD133-positive cells show CSCs-specific self-renewal properties. It was determined that the CD133 gene expression difference between CD133-positive CSCs and CD133-negative cells was approximately 25-fold. It has been observed that CD133-positive CSCs express more pluripotency genes (such as OCT-4, SOX-2 and KLF-4) compared to CD133-negative cells.