PDGF-B链基因的三链形成寡核苷酸对大鼠脑胶质瘤生长及细胞周期的影响  

Effect of triplex forming oligonucleotide of PDGF-B chain on tumor growth and cell cycle of rat glioma

作　　者：李维方[1] 周定标[2] 余新光[2] 金由辛[3] 

机构地区：[1]解放军第306医院神经外科,北京100101 [2]解放军总医院神经外科,北京100853 [3]中国科学院上海生命科学研究院生物化学与细胞生物学研究所分子生物学国家重点实验室,上海200032

出　　处：《第二军医大学学报》2004年第9期966-969,共4页Academic Journal of Second Military Medical University

基　　金：全军医药科研"十五"规划课题 (0 1 MB0 66)

摘　　要：目的 :观察血小板衍生长因子 (platelet- derived growth factor,PDGF) B链基因的三链形成寡核苷酸 (triplex form ingoligonucleotide,TFO)对大鼠胶质瘤细胞生长及细胞周期的影响。方法 :所有 SD大鼠 (36只 ,体质量 2 2 0～ 2 5 0 g)均在右尾状核区微量灌注 1× 10 6个 C6 胶质瘤细胞 ,其中实验 组和 组 (n=12 )在细胞接种后的第 8天开始分别原位注射 TFO1.5mg/ 2 0 μl和 3.0 m g/ 2 0 μl的生理盐水 ,以后每隔 72 h注射相同剂量的 TFO,共注射 3次。对照组 (n=12 )在相同时间原位注射 2 0 μl生理盐水。实验 3周时处死所有的大鼠 ,观察肿瘤的生长情况 ,标本做常规 H- E染色。流式细胞仪检测肿瘤 PDGF- B、PCNA的表达及细胞周期。结果 :实验 组、 组的成瘤抑制率分别为 6 6 .1%、91.8% ,两者相比有显著差异 (P<0 .0 1)。TFO对胶质瘤细胞 PDGF- B、PCNA表达有明显的抑制作用 (P<0 .0 1) ;TFO能明显降低胶质瘤细胞 G2 - M期和 S期 (DNA合成期 )的百分率 ,阻止细胞由静止期 (G0 - G1 期 )进入 DNA合成期 (S期 ,P<0 .0 1)。以上作用均存在浓度依赖性。结论 :原位应用TFO能抑制 PDGF- B表达 ,阻碍细胞进入 S期 ,使胶质瘤细胞增殖降低 ,达到抗胶质瘤生长的作用。Objective:To observe the effect of triplex forming oligonucleotide(TFO) of PDGF-B chain on cell proliferation and cell cycle of rat glioma.Methods:1×10 6 C 6 glioma cells with high-flow microinfusion were seeded into right caudate putamen of rats.TFO was used in situ a week after glioma cell inoculation.The treatment groupⅠandⅡwere treated with 1.5 mg/20 μl and 3.0 mg/20 μl TFO respectively on 8,11 and 14 d after cell inoculation.The control group was treated with 20 μl normal saline with the same procedure.Three weeks after cell inoculation all rats were killed and samples were detected with macroscopic,microscopic histology and immunofluorescence flow cytometric analysis.Results:The inhibition rates of tumor growth were 66.1% in the treatment groupⅠand 91.8% in the treatment group Ⅱ.TFO specifically blocked expressions of PDGF-B and PCNA of glioma cells in a concentration-dependent fashion. TFO obviously inhibited the transit of cells from the G 0-G 1 to S phase in a concentration-dependent fashion.Conclusion:TFO can commendably block PDGF-B expression,inhibit cell proliferation and induce C 6 glioma cells arrest in the G 1 phase,and thus inhibit tumor growth of glioma.

关 键 词：胶质瘤 血小板源生长因子 三链形成寡核苷酸 细胞增殖 细胞周期 

分 类 号：R730.264[医药卫生—肿瘤]