Immunophenotypic characterization of sphere-forming cells derived from the human renal cell carcinoma cell line 786-O.

Cancer stem cells (CSCs) have been found in many different types of malignant tumors. In our previous study, we found that sphere-forming cells (SFCs) from the renal cell carcinoma (RCC) cell line SK-RC-42 are rich in CSCs. However, our previous reports are based on only one human RCC cell line, which makes it difficult to determine whether the findings from this cell line represent the general mechanisms in human RCC. Therefore, in this study, we attempted to evaluate whether the sphere culture method could enrich for CSCs from another human RCC cell line, 786-O, and to characterize their immunological phenotype. We discovered that a small population of 786-O cells was capable of growing as tumor spheres. The SFCs had many properties similar to CSCs, including greater ability to self-renew in vitro and in vivo, higher mRNA expression levels of several 'stemness' genes, stronger tumorigenicity and resistance to chemotherapeutic agents than monolayer adherent cells (MACs). The SFCs expressed low levels of MHC-I, HLA-DR, CD80, CD86, CD152 and CD154. Additionally, the SFCs had lower expression levels of Her2 and hTERT, FasL, Fas, the transcription factor forkhead box protein 3 (FoxP3) and activating natural killer cell receptors than did the MACs. In addition, both 786-O SFCs and MACs were weakly positive for B7-H4 expression, while the expression level of B7-H1 in 786-O SFCs was lower than that in MACs. Furthermore, 786-O SFCs and MACs both expressed substantial and comparable levels of membrane complement regulatory proteins (mCRPs). Finally, we found that 786-O SFCs triggered T cell apoptosis. These findings suggested that tumor spheres from 786-O cells are rich in CSCs. The immunological phenotype of the SFCs described in our study suggests that CSCs might play an important role in tumor immune evasion.