首页> 中文期刊> 《中国组织工程研究》 >亚砷酸钠对人永生化角质形成细胞株恶性转化的影响

亚砷酸钠对人永生化角质形成细胞株恶性转化的影响

         

摘要

BACKGROUND:Studies have found that sodium arsenite can cause the malignant transformation and tumorigenicity of HaCaT cels, but whether low concentrations of sodium arsenite can cause the malignant transformation is rarely reported. OBJECTIVE:To study the effect of sodium arsenite on the malignant transformation of human immortalized keratinocyte cel lines. METHODS:HaCaT cels were treated with different concentrations of sodium arsenite. MTT assay was used to determine the effect of sodium arsenite on HaCaT cel morphology and proliferation, flow cytometry used to detect the effect of sodium arsenite on HaCaT cel cycle, and soft agar colony formation experiments assay used to determine the effect of sodium arsenite on HaCaT cel colony formation capacity. RESULTS AND CONCLUSION: HaCaT cels grew wel when the concentration of sodium arsenite was 5 mol/L, but the cel growth was inhibited under intervention with 10 and 50 mol/L sodium arsenite. HaCaT cels treated with 0.1 mol/L sodium arsenite were passaged to the 20th generation, and cel morphology had no notable changes; cels at passage 25 exhibited enlarged size and multiple nucleoli, which had a continued proliferation trend. Compared with the primarily cultured cels, 0.1 mol/L sodium arsenite-treated HaCaT cels at passages 15 and 25 had an increased proportion at S phase and G2/M phase, with strengthened proliferation ability and increased colony-forming efficiency, and moreover, the proliferation ability and colony-forming efficiency of passage 25 cels were higher than those of passage 15 cels. These experimental data show that high concentrations of sodium arsenite reduce HaCaT cel viability, and low concentrations of sodium sulfite have a certain influence on the morphology, cel cycle, proliferation ability and colony-forming efficiency of HaCaT cels, and moreover, the proliferation ability and colony-forming efficiency of human immortalized keratinocytes wil be strengthened with the increase of passage.%背景:有研究发现亚砷酸钠可以引起HaCaT细胞的恶性转化,并具有致瘤性,但关于低浓度亚砷酸钠是否引起HaCaT细胞发生恶性转化的研究不多。  目的:分析亚砷酸钠对HaCaT细胞恶性转化的影响。  方法:将HaCaT细胞加入不同浓度的亚砷酸钠进行培养,MTT测定亚硫酸钠对HaCaT细胞形态和增殖能力的影响,流式细胞术测定亚硫酸钠对HaCaT细胞周期的影响,软琼脂集落形成实验测定亚硫酸钠对HaCaT细胞克隆形成能力的影响。  结果与结论:①5 mol/L亚砷酸钠对HaCaT细胞生长没有影响,但10和50 mol/L亚砷酸钠抑制HaCaT细胞生长。②HaCaT细胞经0.1 mol/L亚砷酸钠处理后,传代培养至第20代时,HaCaT细胞的形态没有明显改变,第25代时,细胞体积增大,有多个核仁,有不断增殖趋势。③与原代细胞相比,经0.1 mol/L亚砷酸钠处理后传至第15及25代HaCaT细胞S期细胞和G 2/M期细胞所占比例增加,增殖能力增强,克隆形成率增加,且第25代HaCaT细胞的增殖能力和克隆形成率高于第15代。④实验结果说明,高浓度亚砷酸钠降低HaCaT细胞的活力,低浓度亚硫酸钠对HaCaT人永生化皮肤角质形成细胞的细胞形态、细胞周期、增殖能力以及克隆形成能力均有一定的影响,随着传代的增加,人永生角质形成细胞增殖能力和克隆形成能力不断增强。

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