PDGF-B链基因的TFO对大鼠脑胶质瘤细胞增殖和凋亡影响  

The effect of triplex forming oligonucleotide of PDGF-B chain on cell proliferation and cell apoptosis 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306医院神经外科 [2]解放军总医院神经外科 [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

出　　处：《肿瘤》2005年第1期19-23,共5页Tumor

基　　金："十五"军队科研基金项目(编号:01MB066)

摘　　要：目的　观察原位注射血小板源生长因子(platelet derivedgrowthfactor,PDGF)B链基因的三链形成寡核苷酸(Triplexformingoligonucleotide,TFO)对荷瘤大鼠胶质瘤细胞增殖和凋亡影响。方法　所有大鼠均在立体定向导引下右尾状核区微量灌注1×106C6胶质瘤细胞,其中实验Ⅰ和Ⅱ组在细胞接种后的第8天开始分别在肿瘤部位注射TFO 1. 5mg/20μL和3.0mg/20μL的生理盐水,以后每隔72h原位注射相同剂量的TFO,共注射3次。对照组仅在相同时间原位注射20μL生理盐水。实验至3周时处死所有的大鼠,观察肿瘤的生长情况,定性和定量观察肿瘤PDGF B的表达、PCNA表达及细胞凋亡。结果　实验Ⅰ组的成瘤抑制率为66.1%,实验Ⅱ组为91.8%。TFO对C6胶质瘤细胞PDGF B、PCNA表达有明显的抑制作用;TFO能使C6胶质瘤细胞凋亡增加,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,使胶质瘤细胞增殖降低、细胞凋亡增加,TFO能取得很好的抗胶质瘤生长的治疗效果。Objective To observe the effect of triplex forming oligonucleotide (TFO)of PDGF-B chain.on cell proliferation and cell apoptosis of rats glioma. Methods 1×10 6 C 6 glioma cells with high-flow microinfusion were seeded into right caudate putamen of all rats with stereotactic technique. TFO was used in situ with stereotactic technique at day 8 after glioma cell inoculation. The treated groupsⅠandⅡwere treated with 1.5mg/20μl and 3.0mg/20μl TFO 3 times at days 8,11 and 14 after cell inoculation,respectively. The control groupⅠwas just treated with 20μl saline 3 times at the same time. Three weeks after cell inoculation,all rats were killed.Samples were detected with macroscopic and microscopic histology as well as immunofluorescence flow cytometric analysis. Results The inhibition rates of tumor growth were 66.1% in the treated groupⅠ, 91.8% in the treated group Ⅱ. PDGF-B and PCNA expressions of qualitative analyses of C 6 glioma cells with immunohistochemistry were weak in treated groupⅠand in the treated group Ⅱ, strong in the control group. PDGF-B and PCNA expressions of quantitative analyses were detected by immunofluorescence flow cytometric technique of C 6 glioma cells. From the flow cytometric analysis,it was demonstrated that TFO could specifically block these expressions of C 6 glioma cells in concentration-dependent fashion. TFO could significantly induce glioma cell apoptosis in concentration-dependent fashion. Conclusion PDGF-B is required for the maintenance and the development of tumor growth of glioma. TFO used in situ could apparently block PDGF-B expression,inhibit cell proliferation and induce cell apoptosis in order to inhibit tumor growth of glioma well.

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

分 类 号：R730.264[医药卫生—肿瘤]