Objective To evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro. Methods DNA strand breakage was determined by sin...Objective To evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro. Methods DNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay. Results Exposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC. Conclusions GMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.展开更多
基金This work was supported by a grant from the National Natural Science Foundation of China (grant no. 39840017).
文摘Objective To evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro. Methods DNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay. Results Exposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC. Conclusions GMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.
文摘目的:分析甲基丙烯酸环氧丙酯(glycidyl methacrylate,GMA)致人支气管上皮16HBE细胞恶性转化过程中不同时点甲基化转移酶及甲基CpG结合蛋白家族基因的表达差异,初步探讨GMA诱导16HBE细胞发生恶性转化的表观遗传机制。方法:采用人类全基因组表达谱芯片筛选GMA诱导16HBE细胞恶性转化过程中甲基化转移酶及甲基CpG结合蛋白家族的差异表达基因,并采用实时定量PCR(Real time PCR)技术检测筛选所得差异基因DNMT1及MBD1 mRNA的表达情况。结果:芯片结果显示在转化第10代(前期)细胞中DNMT1及MBD1基因较同代龄对照细胞表达上调,实时定量PCR进一步确证DNMT1及MBD1基因表达分别上调80.60%、79.10%,与芯片结果一致。结论:甲基化可能是GMA致16HBE细胞发生恶性转化的一种机制,DNMT1及MBD1基因可能是多个甲基化相关基因转录表达下调的关键调控点,其在GMA致16HBE细胞恶性转化过程中起着重要作用。
