联合VEGF反义寡核苷酸和PDGF三链形成寡核苷酸抑制大鼠脑胶质瘤生长  

Triplex forming oligonucleotide of PDGF-B chain combined with antisense oligonucleotide of VEGF inhibits glioma growth in rats

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》2006年第2期160-164,共5页Academic Journal of Second Military Medical University

基　　金：全军医药科研"十五"规划课题(01MB066)~~

摘　　要：目的:观察血小板源生长因子(PDGF)B链基因(PDGF-B)的三链形成寡核苷酸(triplex forming oligonucleotide,TFO)PDGF-TFO和血管内皮生长因子(VEGF)反义寡核苷酸(antisense oligonucleotide,AON)VEGF-AON对大鼠脑胶质瘤生长的抑制作用。方法:36只雄性SD大鼠,分为4组,所有大鼠均在立体定向导引下行右尾状核区微量灌注含1×106C6胶质瘤细胞的生理盐水20μl。在细胞接种后第8天,实验Ⅰ组6只大鼠原位注射含PDGF-TFO 1.5 mg的20μl生理盐水,实验Ⅱ组12只和实验Ⅲ组12只大鼠则分别原位注射含PDGF-TFO 1.5 mg+VEGF-AON 0.125 mg和PDGF-TFO 1.5 mg+VEGF-AON 0.25 mg的20μl生理盐水。以后每隔72 h原位注射相同剂量的药物1次,共注射3次。对照组6只大鼠仅在相同时间原位注射20μl生理盐水。实验3周时处死所有大鼠,观察肿瘤的生长情况,定性和定量观察肿瘤PDGF-B、VEGF和肿瘤核增殖抗原(PCNA)表达。结果:实验Ⅰ组的成瘤抑制率为53.1%,实验Ⅱ组为81.4%,实验Ⅲ组为93.1%,3组比较有明显差异(P<0.01)。PDGF-TFO对C6胶质瘤细胞PDGF-B、VEGF、PCNA表达有明显的抑制作用;联合应用PDGF-TFO和VEGF-AON能更好地抑制PDGF-B、VEGF、PCNA表达。结论:联合应用PDGF-TFO和VEGF-AON比单用PFGF-TFO能更有效地抑制肿瘤生长。Objective：To observe the inhibitory effect of triplex forming oligonucleotede （TFO） of PDGF-B chain combined with antisense oligonucleotide （AON） of VEGF on glioma growth in rats. Methods： Thirty-six male SD rats received 1 × 10^6 C6 glioma cells/20μl normal saline into the right caudate putamen by stereotactic technique. Group Ⅰ （n = 6） was treated in situ with 1.5 mg/20μl TFO for 3 times on 8,11 and 14 d after glioma cell inoculation. Group Ⅱ （n = 12） and Ⅲ （n = 12） were respectively treated with TFO 1.5 mg＋AON 0. 125 mg/20μl and TFO 1.5 mg＋AON 0. 250 mg/20μl for 3 times on 8,11 and 14 d after glioma cell inoculation. Control group （n=6） was treated with 20 /11 saline for 3 times at the same time pionts. Three weeks after cell inoculation,all rats were sacrificed to observe tumor growth and to determine the expression of PDGF-B,VEGF and PCNA. Results： The inhibition rate of tumor growth was 53.1% in group Ⅰ , 81.4% in group Ⅱ and 93.1% in group Ⅲ , with significant difference found between the 3 groups （P〈0. 01）. PDGF-TFO had obvious inhibitory effect on the expression of PDGF-B,VEGF and PCNA in C6 glioma cells;and PDGF-TFO combined with VEGF-AON had more potent inhibitory effect on the expression of PDGF-B, VEGF and PCNA in Ce glioma cells. Conclusion： PDGF-TFO combined with VEGF-AON has more potent inhibitory effect on tumor growth than PDGF-TFO alone.

关 键 词：胶质瘤 血小板源生长因子 三链形成寡核苷酸 血管内皮生长因子 反义寡核苷酸 

分 类 号：R739.41[医药卫生—肿瘤]