灵芝酸A对人胶质瘤细胞U251细胞增殖、凋亡和侵袭的影响

目的探讨灵芝酸A对人胶质瘤细胞U251细胞凋亡、侵袭及KDR表达的影响。方法制备灵芝酸A,以人胶质瘤细胞细胞U251细胞为研究对象,根据细胞培养液所含灵芝酸A的不同浓度,将实验分为空白对照组(等量细PBS)、灵芝酸A低浓度组(0.1 mmol/L)和高浓度组(0.5 mmol/L)。用RT-PCR和Western blot法分别从mRNA水平和蛋白水平检测KDR基因的表达,CCK-8法测定细胞体外增殖能力,流式细胞技术(flowcytometry,FCM)检测各组细胞周期和凋亡情况,TUNEL染色检测各组细胞的的凋亡,细胞侵袭小室法检测各组细胞的体外侵袭力。结果 RT-PCR和Western blot显示,灵芝酸A高浓度组和低浓度组较空白对照组的KDR mRNA和蛋白表达都明显下降;灵芝酸A高浓度组和低浓度组细胞的生长速度明显减慢,降低G1期细胞比例,S期和G2/M期比例增高;与空白对照组相比,高浓度组和低浓度组细胞的凋亡率明显升高(P0.01),增殖、侵袭能力显著下降(P0.05);高浓度组和低浓度组细胞相比促进凋亡和抑制KDR表达的作用更明显(P0.05)。结论灵芝酸A可诱导人胶质瘤细胞U251细胞凋亡,抑制其增殖和侵袭能力,并能抑制Kdr DR mRNA和蛋白的表达,提示这可能是其抗肿瘤的作用机制之一。

Effect of Ganoderma acid A to human glioma cells U251 cells on proliferation, apoptosis and invasion

Objective To investigate the effect of ganoderic acid A(GA-A) on apoptosis, invasion and KDR expression of human U251 cells. Methods Ganoderic acid A(GA-A) was prepared, human U251 cells were treated with 0.1, and 0.5 mmol/L GA-A, and the experiment was divided into blank control, low concentration and high concentration group. The expressions of KDR mRNA and KDR protein was assayed by RT-PCR and Western blot.The effect of GA-A on the proliferation and invasion capability of U251 cells was determined by CCK-8 and transwell assay in vitro, respectively. Flow cytometry was used to detect the influence of GA-A on the cell cycle and apoptosis of U251 cells, and TUNEL staining was detected the cell apoptosis too. Results Compared with the control group, KDR mRNA and protein expression of high concentration and low concentration group were significantly decreased(P < 0.05), GA-A can significantly reduce the cell growth rate, reduce the proportion of cells in G1 phase and increase the proportion of S phase and G2/M phase, cells apoptosis was significantly increased in the high concentration and low concentration group (P < 0.01), and cells proliferation and invasion was significantly decreased (P < 0.05). Compared with low concentration group, the high concentration group induce cell apoptosis and inhibit the expression of KDR more significant (P < 0.05). Conclusions Ganoderma acid A can induce apoptosis in U251 cells, inhibit proliferation and invasion, and can inhibit the expression of KDR mRNA and protein, which may be one of the mechanisms of anti-tumor.