Nickel-induced transformation of human cells causes loss of the phosphorylation of the retinoblastoma protein.
The retinoblastoma (Rb) protein (pRb) has been studied in various crystalline NiS-transformed cell clones derived from the human osteoblast cell line, HOS TE-85. The parental HOS cells were not able to proliferate in soft agar medium, but they acquired this property following treatment with crystalline NiS. The pRb was found only in the hypophosphorylated form in 8 of 9 nickel-transformed clones examined, whereas in the parental cells the pRb appeared in both phosphorylated and unphosphorylated forms. Neither Rb gene expression nor its phosphorylation was affected by acute nickel treatments of HOS cells. The nickel-transformed HOS clones expressed the major regulators of Rb phosphorylation, cyclin E and cdk-2, at levels similar to those of the parental cells. In coimmunoprecipitation assays with cell lysates from the transformed clones that exhibited the hypophosphorylated form of pRb, the Rb protein failed to form a complex with simian virus 40 large T-antigen, indicating a lack of functional activity. When a plasmid containing the normal Rb gene was transfected into these nickel-transformed cells, it restored the Rb phosphorylation pattern observed in parental cells and the cells acquired a normal phenotype (i.e., they were no longer able to grow in soft agar). This suggested that a mutation was induced in nickel-transformed cells that affected the ability of the Rb protein to be phosphorylated and function normally, and this mutation allowed the human nickel-transformed cells to acquire anchorage-independent growth.
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Nelson Institute of Environmental Medicine, New York University Medical Center, New York 10016.
Dowjat
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Costa
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D000937:Antigen-Antibody Reactions; D000957:Antigens, Viral, Tumor / Q000378:metabolism; D002461:Cell Line, Transformed; D006801:Humans; D009532:Nickel / Q000633:toxicity*; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D016160:Retinoblastoma Protein / Q000187:drug effects* / Q000378:metabolism; D013539:Simian virus 40 / Q000276:immunology
D000957:Antigens, Viral, Tumor; D016160:Retinoblastoma Protein; D009532:Nickel