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Alterations of human endometrial stromal cells produced by N-methyl-N'-nitro-N-nitrosoguanidine
Abstract:
Stromal cell cultures obtained from human endometrium were treated repetitively with N-methyl-N'-nitro-N-nitrosoguanidine in vitro at concentrations ranging from 0.5 to 4.0 micrograms/ml, and alterations in growth potential and morphology were analyzed. A single exposure to the carcinogen resulted in morphological evidence of toxicity and reductions in growth rates, plating efficiency, and saturation density as compared to solvent-treated control cells. Cytotoxicity was reduced after additional exposures to the carcinogen. Following repetitive treatments with N-methyl-N'-nitro-N-nitrosoguanidine, human endometrial stromal cells developed enhanced growth potential, the capacity to form macroscopic colonies in soft agar, and elevated gamma-glutamyltranspeptidase activity. Carcinogen-treated cells displayed atypical morphology characterized by irregularities in cell and nuclear size and shape, large bizarre nucleoli, increased nuclear:cytoplasmic ratios, and cellular crowding. Control cells did not display altered morphology or growth parameters even following multiple exposures to solvent and repetitive subculturing. These alterations in growth potential and morphology suggest that the cells are progressing towards preneoplastic and perhaps neoplastic transformation in vitro.
Insights
Human endometrial stromal cells exposed to a carcinogen showed changes indicating preneoplastic transformation. These cells developed enhanced growth and atypical morphology after repeated N-methyl-N'-nitro-N-nitrosoguanidine treatment.
Area of Science:
- Cell biology
- Carcinogenesis research
- Gynecologic oncology
Background:
- Human endometrial stromal cells (hESCs) are crucial for uterine function.
- Understanding cellular transformation is key to cancer research.
- N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a known mutagen and carcinogen.
Purpose of the Study:
- To investigate the in vitro effects of repetitive N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) exposure on human endometrial stromal cells (hESCs).
- To analyze alterations in growth potential and morphology indicative of cellular transformation.
Main Methods:
- Primary hESCs were cultured and treated with varying concentrations of MNNG.
- Morphological changes, growth rates, plating efficiency, and soft agar colony formation were assessed.
- Gamma-glutamyltranspeptidase activity was measured.
Main Results:
- Single MNNG exposure induced toxicity, reducing growth rates and plating efficiency.
- Repeated MNNG treatment led to enhanced growth potential, soft agar colony formation, and elevated gamma-glutamyltranspeptidase activity.
- Carcinogen-treated cells exhibited atypical morphology, including irregular cell/nuclear size, large nucleoli, and increased nuclear:cytoplasmic ratios.
Conclusions:
- Repetitive MNNG treatment induces significant alterations in hESCs, suggesting progression towards preneoplastic and potentially neoplastic transformation in vitro.
- These findings provide a model for studying endometrial carcinogenesis.
- The observed cellular changes highlight the potential of MNNG to induce transformation in endometrial stromal cells.