Isolation and characterization of cancer stem-like cells from ACHN renal cancer cells / 中国医师杂志

ABSTRACT

Objective To isolate and identify stem-like cells (CSCs) from human ACHN renal cell carcinoma cell line.Methods ACHN cancer stem cells CSCs were enriched by serum-free culture condition.Methyl thiazolyl tetrazolium (MTT) method was performed to draw the growth curve.Flow cytometry was performed to evaluate the expressions of two CSCs markers,CD133 and CD44.Real-time polymerase chain reaction (RT-PCR) was performed to evaluate the expression of stem-related genes (Oct3/4,Bmi,Nanog,and 3-catenin).The tumorigenicity in vivo was also examined.Results ACHN CSCs were formed by sphere-forming culturing.The data demonstrated that there were stronger capacities of proliferation in ACHN tumor spheres than that of ACHN cells.And the expressions of CD133 and CD44 were also significandy enhanced in tumor spheres.The data showed that four genes (Oct3/4,Bmi,Nanog,and β-catenin)were upregulated in sphere-forming cells.Sphere-forming cells had a higher tumorigenic potential in vivo than incubated cells.Conclusions Human ACHN renal cell carcinoma cell line sphere-forming cells can be enriched by serum-free culture condition and exhibits several characteristic cancer stem cells properties.