壳寡糖对Hela细胞cyclin D1、bcl-xl和bcl-2 mRNA表达的影响

背景：研究表明壳寡糖具有抗肿瘤作用，但壳寡糖对cyclin D1、bcl-2和bcl-xl影响机制尚不清楚。 目的：观察壳寡糖抑制Hela细胞生长的作用，以及对细胞内cyclin D1、bcl-xl和 bcl-2 mRNA表达的影响。 方法：壳寡糖(0.1，0.5，1，2，5 g/L)刺激Hela细胞后，CCK8实验检测其对细胞生长的影响；运用实时荧光定量PCR方法在分子水平检测cyclin D1、bcl-xl和 bcl-2 mRNA在细胞中的表达情况。 结果与结论：壳寡糖可抑制Hela细胞的生长；随着壳寡糖的质量浓度由0.1 g/L增加至2 g/L，壳寡糖抑制cyclin D1，bcl-2，bcl-xl基因表达的作用也逐渐增强，在2 g/L时作用最强，但质量浓度过高时(5 g/L)抑制作用反而有所减弱。结果表明壳寡糖的抗肿瘤作用可能通过两方面进行：一方面壳寡糖可以抑制与细胞周期有关的cyclin D1 mRNA表达以抑制肿瘤细胞的增殖；另一方面壳寡糖可以抑制抗凋亡蛋白Bcl-2 和bcl-xl mRNA的表达以促进肿瘤细胞的凋亡；且对前者的作用强于后者。 关键词：荧光定量PCR；壳寡糖；Hela细胞；细胞周期蛋白 D1；bcl-xl；bcl-2 doi:10.3969/j.issn.1673-8225.2010.03.012

Effect of chitooligosaccharides on cyclin D1, bcl-xl and bcl-2 mRNA expression in Hela cells

BACKGROUND: Studies have shown that chitooligosaccharides have antitumor effect. However, the influence of chitooligosaccharides on cyclin D1, bcl-2 and bcl-xl remains unclear. OBJECTIVE: To observe the inhibition effect of chitooligosaccharides on the proliferation of Hela cells, and the influence on cyclin D1, bcl-xl and bcl-2 mRNA expression. METHODS: Hela cells were stimulated by different concentrations of chitooligosaccharides (0.1, 0.5, 1, 2, 5 g/L). The effects of chitooligosaccharides on Hela cells were detected by CCK8 kit. Using real-time PCR methods, the gene expression of cyclin D1, bcl-xl and bcl-2 mRNA were determined. RESULTS AND CONCLUSION: Chitooligosaccharides inhibited the proliferation of Hela cells. With the concentrations of chitooligosaccharides increased from 0.1 g/L to 2 g/L, the inhibition effects on the gene expression of cyclin D1, bcl-xl and bcl-2 were enhanced, peaked at 2 g/L, and decreased at high concentration (5 g/L). Antitumor activity of chitooligosaccharides may exert through two aspects: it inhibits cyclin D1 mRNA expression to suppress the proliferation of tumor cells; on the other hand, chitooligosaccharides inhibits the expression of bcl-xl and bcl-2 to promote the apoptosis of tumor cells. Moreover, the effects of the former are stronger than the latter.