Establishment and characterization of seven human renal cell carcinoma cell lines

Objective To establish human renal cell carcinoma (RCC) cell lines, and to investigate the cell phenotypes and molecular characteristics of human RCC cell lines and their corresponding tumour tissues. Materials and methods Seven human RCC cell lines from pathologically proven RCCs were established. The histopathology of the primary tumours, in vitro growth characteristics and status of tumour suppressor genes, mismatch rep air genes and microsatellite instability (MSI) were examined in cell lines and their corresponding tumour tissues. Five of the cell Lines were derived from clear cells (SNU-228, -267, -328, 349, and -1272), one from granular cells (SNU-482), and one from mixed clear and granular cell types (SNU-333) . The mutational status was compared for von Hippel-Lindau (VHL), p53, TGP- beta type II receptor (TGF-beta RII), hMSH2, and hMLH1 genes in the cell li nes and their corresponding tumour tissues. The MSI status of the cell line s was determined by screening for adenine repeat sequences, e.g. BAT-25, BA T-26, and BAT-40. Results All lines showed different doubling times and were confirmed by DNA fingerprinting analysis to be unique. Contamination by mycoplasma or bacte ria was excluded. In two cell lines (SNU-349 and -1.272) and their tumour t issues, mutations in the VHL gene were found. The SND-267 line had a frames hift mutation in the p53 gene. A missense mutation of the TGF-beta RII gene was detected in the SNU-1272 line and the corresponding tissue. Analysis o f the repeat sequences showed one cell line (SNU-349) to have MSI and the o ther six to have microsatellite stability, As MSI is a hallmark of the inac tivation of mismatch repair genes, the presence of hMSH2 and hMLH1 mutation s was investigated in all seven cell lines. An inactivating homozygous sing le base-pair deletion of the hMLH1 gene was found only in the SNU-349 cell line and corresponding tissue. Moreover, a frameshift mutation within an 8- bp polyadenine repeat present in the hMSH3 coding region was found only in the MSI cell line and tumour tissue. Conclusion These newly established RCC cell lines should provide a useful i n vitro model for studies related to human RCC, The SNU-349 cell line shoul d be especially useful for studies of MSI and mismatch repair-defective RCC s.