欢迎访问《现代生物医学进展》编辑部！

目的：观察全反式维甲酸（ATRA）处理和ATRA 与十四烷酰佛波醇乙酸酯（TPA）序贯处理（ATRA/TPA）对人类神经母细胞瘤细胞系SH-SY5Y 细胞增殖抑制和形态分化的影响。方法：应用10μM ATRA 处理6 天和10μM ATRA 处理3 天继以80 nM TPA 处理3 天这两种方法使SH-SY5Y 细胞分化；用倒置光学显微镜动态观察SH-SY5Y 细胞形态学变化；并用MTT 比色法比较两种分化方法对SH-SY5Y 细胞的体外抗增殖作用。结果：ATRA 处理和ATRA 与TPA 序贯处理对SH-SY5Y 细胞都有抗增值和诱导细胞分化作用，细胞形态发生明显的变化，分化成神经元表型，前者主要表现为两端带有长突起的纺锤体样细胞形态，而后者主要是由细胞体延伸出多个突起的多边形的细胞。ATRA 分化6 天的细胞的存活率下降为78.7%±2.0%。当去除ATRA 后，继续培养1 天的细胞存活率上升为89%±0.2%，而继续培养2 天的细胞存活率为86.3%±1.4%；ATRA 与TPA 序贯分化6 天细胞存活率下降为75.9±0.4%。当去除TPA 后，继续培养一天的细胞存活率为75.5±0.7%，继续培养2 天的细胞存活率为74.9± 1.0%。结论：维甲酸（ATRA）处理和ATRA 与十四烷酰佛波醇乙酸酯（TPA）序贯处理（ATRA/TPA）均能明显诱导SH-SY5Y 细胞分化。这两种分化细胞为神经科学的研究提供了优良的体外培养模型细胞，尤其是ATRA 与TPA 序贯处理能获得分化完全而稳定的神经元样细胞。

英文摘要:

Objective: To investigate the effects of All trans-retinoic acid (ATRA) and ATRA followed by phorbol ester 12-o-tetradecanoylphorbol- 13-acetate (TPA) (ATRA/TPA) on growth and morphological differentiation of human neuroblastoma SH-SY5Y cell line. Methods: SH-SY5Y cells were differentiated with 10 μM ATRA for 6 days and 10 μM ATRA for 3 days followed by 80 nM TPA for another 3 days (RA/TPA). Morphological differentiation was detected by using the inverse microscope. And MTT assay was used to detect the anti-proliferative effect of ATRA and ATRA/TPA on SH-SY5Y cells. Resulls: Both ATRA and ATRA/TPA could induce morphological differentiation and inhibit growth of SH-SY5Y cells. Differentiated cells underwent striking morphological changes and resulted in neuronal phenotype. ATRA-differentiated SH-SY5Y cells exhibited a spindle-shaped morphology with elongated processes extending from opposing ends. Whereas ATRA/TPA-differentiated SH-SY5Y cells showed polygon-shaped morphology with numerous elongated processes extending from cell body. The cell viability decreased to 78.7%±2% when exposured to ATRA at the concentration of 10μmol/ L for 6 days. After the removal of ATRA, cell viability increased to 89%±0.2% and 86.3%±1.4%, respectively, when cultured for another one day and two days. The cell viability decreased to 75.9±0.4% when differentiated with 10 μM ATRA for 3 days followed by 80 nM TPA for another 3 days. The cell viability was 75.5±0.7% and 74.9±1%, respectively, when SH-SY5Y cells were stoped treating with ATRA/TPA and cultured for another one day and two days. Conclusion: ATRA and ATRA/TPA induced SH-SY5Y cell differentiation. The two kind of differentiated SH-SY5Y cell provided excellent in vitro cell model for neurological studies, especially RA/TPA differentiated SH-SY5Y cells.

查看全文查看/发表评论下载PDF阅读器

关闭