Transformation of C3H/10T 1/2 mouse embryo cells to focus formation and anchorage indepence by insoluble lead chromate but not soluble calcium chromate: Relationship to mutagenesis and internalization of lead chromate particles

The genotoxicity of soluble and insoluble hexavalent chromium compounds was studied in mammalian cell assays which detect base substitution, deletion, addition, and frameshift mutations [6-thioguanine resistance in Chinese hamster ovary cells], primarily base substitution mutations [ouabain resistance in Chinese hamster ovary and C3H/10T 1/2 Cl 8 mouse embryo fibroblasts (10T 1/2 )] and morphological transformation [focus formation] in 10T 1/2 cells. Soluble hexavalent CaCrO4, administered in either acute (5-h) or subacute (24-h) dosing regimens, induced dose-dependent cytotoxicity and mutation to 6-thioguanine resistance in Chinese hamster ovary cells but no mutation to ouabain resistace or focus formation in transformation assays, although the acute treatment induced a high frequency of conversion of 10T 1/2 cells to adipocytes. Cell lines established from cloned adipocytic cells were not morphologically transformed and did not grow in soft agarose. PbCrO4 did not induce mutationto either 6-thioguanine or ouabain resistance but did induce a reproducible dose-dependent, low frequency of focus formation in 10T 1/2 cells. Cell lines established from PbCrO4-induced foci stably formed foci when coseeded with 10T 1/2 cells, had 3-5-fold increased saturation densities releative to nontransformed 10T 1/2 cells, and formed colonies in soft agarose, indicating their likelihood to be neoplastic. Long term exposure of 10T 1/2 cells to either CaCrO4 or PbCl2, even at 85% cytotoxic concentrations, or pretreatment of cells with either CaCrO4 or PbCl2 followed by treatment with the alternate compound, did not induce morphological transformation. Treatment of cells with insoluble hexavalent PbCrO4 resulted in progressive and extensive vacuolization of cells in contact with the particles. Progressive cytoplasmic engulfment of PbCrO4 particles was observed using scanning electron microscopy, although PbCrO4 particles were not observed inside vacuoles. These results indicated that the soluble clastogens K2Cr2O7 and CaCrO4 were probably mutagenic by a non-base substitution mechanism but could not transform 10T 1/2 cells. In contrast, PbCrO4 was not detectably mutagenic but induced transformation, which could not be explained solely by acute or chronic exposure to dissolution products of either lead or chromate alone. Since PbCrO4 particles were found to be intracytoplasmic in extensively vacuolated cells, we suggest that the unique physicochemical properties of PbCrO4 particles, leading the their internalization and the resultant associated cellular stress response, may be related to the transformation induced by this compound.

Duke Scholars

Author Steven Patierno Medicine, Medical Oncology