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目的:研究黄连素对神经胶质瘤U251细胞增殖、细胞周期的影响,并探讨其抗肿瘤的作用机制。方法:MTT法检测不同浓度黄连素作用U251细胞24 h对其的增殖的影响;流式细胞术检测黄连素对U251的细胞周期阻滞情况;RT-PCR检测黄连素对p21WAF1/CIP1、Cyclin D1的影响。Western blot检测黄连素对p21WAF1/CIP1、Cyclin D1、组蛋白H3/H4的影响。结果:MTT表明黄连素对U251细胞生长的抑制作用呈现剂量依赖性,在1.172、2.344、4.688、9.375、18.750μg/mL的浓度下,抑制率分别为9.211%、19.737%、28.947%、51.974%、84.211%;流式细胞术结果显示,黄连素阻滞U251细胞周期于G2/M期;RT-PCR检测表明其能显著上调U251细胞的p21WAF1/CIP1的表达。Western blot结果表明,黄连素能上调p21WAF1/CIP1表达,而下调Cyclin D1、组蛋白H3/H4表达。结论:黄连素可抑制U251细胞的增殖,并阻滞细胞周期于G2/M期。其抗肿瘤的机制可能与上调p21WAF1/CIP1、下调Cyclin D1表达相关。
OBJECTIVE: To study the effect of berberine on the proliferation and cell cycle of glioma U251 cells and to explore its anti-tumor mechanism. Methods: The proliferation of U251 cells treated with different concentrations of berberine for 24 h was detected by MTT assay. The cell cycle arrest of U251 by berberine was detected by flow cytometry. The expressions of p21WAF1 / CIP1, Cyclin D1 Impact. The effects of berberine on p21WAF1 / CIP1, Cyclin D1 and histone H3 / H4 were detected by Western blot. Results: MTT showed that berberine could inhibit the growth of U251 cells in a dose-dependent manner. The inhibitory rates of berberine were 9.21%, 19.737%, 28.947% and 51.974% at the concentrations of 1.172, 2.344, 4.688, , 84.211% respectively. The results of flow cytometry showed that berberine blocks U251 cell cycle in G2 / M phase. RT-PCR results show that berberine can significantly up-regulate the expression of p21WAF1 / CIP1 in U251 cells. Western blot results showed that berberine can up-regulate the expression of p21WAF1 / CIP1 and down-regulate the expression of Cyclin D1 and histone H3 / H4. Conclusion: Berberine can inhibit the proliferation of U251 cells and arrest the cell cycle at G2 / M phase. The anti-tumor mechanism may be related to up-regulation of p21WAF1 / CIP1 and down-regulation of Cyclin D1 expression.