缓释BCNU对GL261胶质瘤细胞凋亡作用机制的研究

Study on GL261 glioma cell apoptosis mechanisms induced by sustained release BCNU

目的研究BCNU-PLGA对GL261胶质瘤细胞凋亡的作用机制。方法利用溶剂挥发萃取法制备BCNU-PLGA缓释微球。将24只C57BL/6小鼠随机等分为治疗组与非治疗组,借助动物立体定向仪,将体外培养小鼠GL261胶质瘤细胞接种于小鼠颅内,行MR检查掌握颅内成瘤情况。治疗组于术后第2周立体定位植人BCNU-PLGA缓释片剂,观察小鼠生存期,MRI了解瘤体变化。获取标本行HE染色,并行GFAP、Bax、Bcl-2免疫组化检查检测其表达情况,相关数据采用统计学方法进行分析。结果治疗组与对照组生存期有明显差异(P<0.05),治疗组与对照组凋亡相关基因Bcl-2免疫组化检查提示明显差异性(P<0.05),Bax基本无改变,Bax/Bcl-2比值明显增加。GFAP表达为阳性。结论 BCNU-PLGA局部缓释制剂通过下调Bcl-2蛋白表达,上调Bax/Bcl-2的比值,从而使胶质瘤细胞发生凋亡。

Objective To investigate apoptosis mechanism of BCNU-PLGA on GL261 glioma cells. Methods Solvent volatilization extraction chemical strategies was used for preparation of BCNU-PLGA sustained-release microspheres. 24 mouse were randomly and equally divided into treatment group and non-treatment group. GL261 ceils was intracranially injected into C57BL / 6 mouse by using the stereotaxical technology and magnetic resonance imaging （MRI） was followed. BCNU-PLGA sustained-release wafers were implanted in the treatment group 2 weeks after inoculation, observed survival time, and followed-up by MRI. Specimens was gotten. HE staining and immunohistochemical method was used to check the expression of GFAP, Bax and Bcl-2. Statistical methods was used for analysis of relevant data. Results BCNU-PLGA sustained-release wafers were fabricated. Successful creation of C57BL / 6 mouse glioma GL261 model, the ratio was 100%, BCNU-PLGA suatained-release wafer was implanted successfully. The survival time between the treatment group and control groups had obvious difference （ P 〈 0.05 ）. Apoptosis related gene Bcl-2 expression between groups had obvious difference （ P 〈 0.05 ）, wheras no influence on Bax expression, with the ratio of Bax/Bcl-2 increased. The GFAP expression was positive. Conclusion BCNU-PLGA sustained-release microsphere induced glioma apoptosis by its downregulating of Bcl-2 protein expression and up-regulating the ratio of Bax/Bcl-2.