替莫唑胺联合鸦胆子油乳对胶质瘤SKMG-4细胞抑制作用研究

The inhibitory effect of temozolomide combined with Brucea javanica oil on glioma SKMG-4 cell

目的研究替莫唑胺联合鸦胆子油乳对胶质瘤SKMG-4细胞的抑制作用。方法实验分为空白对照组、单独应用替莫唑胺组、单独应用鸦胆子油乳组、替莫唑胺联合鸦胆子油乳组,利用四甲基偶氮唑盐(Methyl thiazolyltetrazolium salt,MTT)比色法检测各组对胶质瘤SKMG-4细胞的增殖抑制影响;利用流式细胞仪(Flowcytometry,FCM)分析各组细胞周期的影响。实时荧光定量PCR分析各组对细胞内B细胞淋巴瘤/白血病(B-cell lymphoma/leukemia-2gene,BCL-2)基因抑制作用。结果 MTT比色法显示联合用药组对SKMG-4细胞具有明显增殖抑制作用,与对照组相比,P<0.05。流式细胞仪分析显示联合用药组中G1期为(74.1±0.2)%,S期为(10.2±0.1)%,与对照组相比,P<0.05,说明联合组可明显将胶质瘤细胞分裂抑制在G1期。实时荧光定量聚合酶链反应(Real-time polymerase chain reaction,RT-PCR)分析联合用药组中BCL-2m RNA表达为(0.026±0.003),低于其他各组,与对照组相比,P<0.05,说明更能抑制BCL-2m RNA表达,从而加速肿瘤细胞凋亡。结论替莫唑胺联合鸦胆子油乳能提高其对胶质瘤的抑制作用。

Objective To explore the inhibitory effect of temozolomide combined with Brucea javanica oil on the gliomaSKMG-4 cell. Methods The subjects were divided into blank control group,temozolomide group,Brucea javanica oil group,and combined treatment group,MTT assay was used to measure the proliferation inhibition of golima SKMG-4 cells in the fourgroups. Flow cytometry was used to measure the change of cell cycle. Real-time PCR method was used to analyze the inhibitionof BCL-2 expression in each group. Results The results of MTT method showed that the proliferation of SKMG-4 cells incombined treatment group was significantly inhibited,compared with the control group（P〈0.05）. Flow cytometry analysisshowed that G1 phase and S phase in combined treatment group were（74.1±0.2）% and（10.2±0.1）% respectively,statisticallydifferent from the control group（P〈0.05）,revealing that the combined group could significantly inhibit the glioma cell divisionin the G1 phase. Real- time PCR analysis showed that expression level of BCL- 2m RNA in combined treatment group was（0.026±0.003）,lower than other groups,compared to the control group（P〈0.05）. These revealed that combined treatmentgroup could better suppress the expression of BCL- 2m RNA,thereby accelerated the apoptosis of tumor cell. Conclusion Temozolomide in combination with Brucea javanica oil emulsion can improve the inhibitory effect on glioma.