不同时机协同给药诱导裸鼠脑胶质瘤细胞凋亡的对照研究  

Control study on U251MG glioma cell apoptosis in the brain of nude mouse with anti-proliferative and anti-infiltrative medicines at different times

作　　者：孙建军[1] 王振宇[1] 钟延丰[2] 杜娟[2] 陈英玉[3] 冯智勇 

机构地区：[1]北京大学第三医院神经外科 [2]北京大学基础医学院病理学系 [3]北京大学人类疾病基因研究中心 [4]解放军61786部队门诊部

出　　处：《中华临床医师杂志（电子版）》2012年第21期6694-6698,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：教育部新教师基金(BSD-09-6-11);北京大学第三医院青年骨干基金(74496-01)

摘　　要：目的对比观察不同时机给予持续低剂量抗肿瘤细胞增殖和抗血管增生药物协同治疗裸鼠U251MG脑胶质瘤抑制脑胶质瘤生长的情况。方法分为两实验组,Ⅰ组,将2.5μl细胞悬液接种至6周龄雄性裸鼠脑白质内,接种次日预防性给药,是口服吲哚美辛(4 mg/kg)和腹腔注射榄香烯(70 mg/kg)协同给药;Ⅱ组,将等量的细胞悬液接种至4周龄雄性裸鼠脑白质内,接种后第20天治疗性协同给药,每组2只裸鼠,共4只裸鼠。给药20 d和30 d时,断椎处死裸鼠,脑标本石蜡切片3μm厚,分别行HE、胶质纤维酸性蛋白(GFAP)、CD34、TUNEL和PDCD-5等染色。结果预防性给药20 d裸鼠脑内未见明显增殖瘤细胞,亦无大量凋亡的瘤细胞,30 d时大量瘤细胞发生凋亡,GFAP呈阴性表达;治疗性给药裸鼠脑内仍可见胶质瘤细胞,20 d部分细胞发生凋亡,30 d大量细胞发生凋亡。结论预防性持续低剂量抗肿瘤增殖和抗血管增生药物协同治疗裸鼠脑胶质瘤,可有效遏制肿瘤的生长;治疗性给药虽能起到遏制肿瘤生长的目的,但治疗效果欠佳。Objective To observe the changes in glioma growth characteristic and apoptosis of tumor cells after low-dose combination of anti-proliferative medicine and cyclooxygenase-2 inhibitor at different times.Methods The U251MG cells in exponential phase of growth were made into 107/ml cell suspension in free-serum 1640 and stored in 37 ℃ incubator.The survival rate of cells was above 95%.The U251MG cells were implanted into the right parietooccipital lobe of nude mouse with a 5 micro liter micro amount sample injector.The number of injected U251MG cells was 5×104 for a mouse.One day or twenty days after the model making,the nude mice were treated with combination of elemene and indometacin,twice a week.The mice were divided into two groups.Group Ⅰ was treated one day,and group Ⅱ twenty days after the model making.Two groups were used as controls,including tumor control and blank control.The mice of tumor control were implanted with U251MG cells,and the mice of blank control implanted with normal sodium.Two control groups treated with normal sodium.The animals were killed on the 20th and 30th days after treatments by breaking cervical vertebra.The fixed brain was made into 3 μm slices by paraffin section.The slices were carried out with HE staining and immunohistochemical staining of glial fibrillary acidic protein(GFAP),CD34,programmed cell death 5(PDCD5)and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL).Results The proliferation of glioma cells was predominant in the tumor control mouse brain.A few immature blood vessels were observed in the tumor 20 days after the implantation.The node of tumor was observed 30 days after the implantation.Several immature blood vessels and a great quantity glioma cells were observed in the mice brain implanted for 40 days.The white matter was infiltrated by bulk glioma cells along with capillary vessel clusters in the mouse brain implanted for 50 days.In groups with preventative combination treatment of the two drugs(treated one day after i

关 键 词：神经胶质瘤 脑胶质瘤 治疗 小鼠 裸 

分 类 号：R739.41[医药卫生—肿瘤]