丙戊酸钠增强大鼠胶质瘤C6细胞放射敏感性的体外实验  

Valproate acid sodium enhancing radiation sensitivity of rat C6 glioma cells in vitro

作　　者：牛俊婕[1] 王晗[1,2] 徐英[1] 周勇[1] 姜玉华[1] 

机构地区：[1]山东大学第二医院肿瘤防治中心,济南250033 [2]山东省济宁市第一人民医院,山东济宁272111

出　　处：《山东大学学报（医学版）》2013年第6期15-19,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(26010105201012)

摘　　要：目的观察丙戊酸钠(VPA)对C6胶质瘤细胞系的体外放射增敏性,为治疗胶质瘤提供实验依据。方法体外常规培养C6胶质瘤细胞,采用不同浓度(0、0.25、0.5、1、2、4 mmol/L)的VPA,作用于C6胶质瘤细胞不同时间(24、48、72 h)后,MTT法检测C6胶质瘤细胞增殖抑制率,选择合适的作用浓度和作用时间;0.5 mmol/L VPA作用C6胶质瘤细胞后,分别给予不同剂量(0、2、4、6、8 Gy)的X射线,克隆形成实验观察其对C6胶质瘤细胞的放射增敏作用;流式细胞术Annexin V-FITC/PI双染法检测不同浓度VPA对C6胶质瘤细胞凋亡率的影响;实验分为对照组、药物组、照射组及联合组,实时定量PCR法检测各组凋亡基因Bcl-2及Bax mRNA的表达。结果VPA对大鼠胶质瘤C6胶质瘤细胞具有增殖抑制作用,且呈浓度、时间依赖性,不同浓度、时间之间比较,差异有统计学意义(P<0.05);VPA在0.5 mmol/L浓度时,对C6胶质瘤细胞毒性较低,且可增加X射线对C6胶质瘤细胞的杀伤作用,放射增敏比为1.30;VPA能够诱导C6胶质瘤细胞凋亡,并且凋亡率随浓度的增高而升高,各浓度之间比较差异有统计学意义(P<0.05);VPA对C6胶质瘤细胞的凋亡作用,在转录水平可下调Bcl-2表达,同时上调Bax的表达。结论 VPA对大鼠胶质瘤C6胶质瘤细胞具有增殖抑制作用,且呈浓度、时间依赖性。低浓度的VPA对C6胶质瘤细胞毒性较低,但可增加X射线对C6胶质瘤细胞的杀伤作用,其放射增敏机制与直接抑制细胞增殖、诱导细胞凋亡、调节凋亡相关基因的表达有关。Objective To observe the radiosensitivity of VPA to C6 glioma cells in vitro, so as to provide experimental evidences to the therapy of cerebral gliomas. Methods Rat C6 glioma cells were cultured commonly in vitro. MTT assays was used to measure the proliferation inhibition rate of C6 glioma cells with VPA in different concentrations （0,0.25,0.5,1,2,4mmol/L） and different durations （24,48,72h） to choose the appropriate concentration and time. C6 cells incubated with 0.5mmol/L VPA were exposed to different doses of X-ray irradiation （IR） （0,2,4,6,8Gy）. Cloning experiments were used to determine the radiosensitizing effect of VPA. Flow cytometry double staining with Annexin V-FITC and propidium iodide （PI） was used to detect the effects of various concentrations of VPA on the apoptosis rate of C6 glioma cells. Trials were divided into the control group, the medication group, the irradiated group and the joint group. Real time PCR was used to detect the expression of Bcl-2 and Bax. Results VPA inhibited the proliferation of rat C6 glioma cells in vitro significantly in a time and concentration dependent manner （P〈0.05）. VPA at the concentration of 0.5mmol/L had a lower toxicity on C6 glioma cells and enhanced the killing effect of X-ray on C6 cells. The sensitization enhancement ratio （SER） was 1.30. VPA induced apoptosis in C6 cells, the apoptosis rate was increasing along with the concentration of VPA.The difference of the apoptosis rate among different concentrations was statistically significant（P〈0.05）. On the transcriptional level translation, after the treatment with VPA, the expression of Bcl-2 decreased, whereas the expression of Bax increased. Conclusion VPA inhibits the growth of rat C6 glioma cells in a dose and time dependent manner. Low concentration of VPA is low-toxic and can enhance the radiosensitivity of C6 glioma cells, and the mechanisms for this action might include directly inhibiting cellular proliferation, inducing apoptosis and regulating the expr

关 键 词：组蛋白去乙酰化酶抑制剂 丙戊酸钠 胶质瘤 C6细胞 放射增敏 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]