Plk1基因为靶的反义寡核苷酸对HepG2肝癌细胞的体内外抗肿瘤作用  

Antitumor Activity of Antisense Oligonucleotides Targeted at Plk1 in HepG2 Cell In Vivo and In Vitro

作　　者：林莉[1] 王升启[2] 管伟[2] 杨秉呼[2] 胡小电[1] 

机构地区：[1]军事医学科学院附属医院肿瘤一科,北京100039 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《癌症》2001年第11期1233-1236,共4页Chinese Journal of Cancer

基　　金：国家重点基础研究发展规划项目(G1998051118);国家863项目

摘　　要：目的:以Plk1基因为靶筛选抗肿瘤药物。方法:根据Plk1mRNA的二级结构设计8条反义寡核苷酸药物,以脂质体介导进行转染,MTT染色计算细胞生长抑制率,从中筛选出一条序列并对其进行优化。最后以最佳序列9号进行体外作用持续时间及裸鼠移植瘤细胞生长抑制率分析。结果:针对5'编码区的5号序列在0.2μmol/L、0.4μmol/L和0.8μmol/L时抑制率分别是59.1%、75.5%和90.7%。它抑制细胞增殖作用最强。5号序列的5'端向内移动3个碱基形成的9号序列的抑制率高于其它3条序列,9号的IC50是0.081μmol/L。终浓度为0.2μmol/L的9号药物经脂质体介导转染给药1次后,抑制活性逐渐增加,第4天抑制率达到93.0%的峰值,然后缓慢下降,到第6天抑制率为78.3%。荷瘤裸鼠每日腹腔注射9号药物(25mg/kg)用药28天,处死时对照组的瘤重为(1.536±0.196)g,用药组瘤重为(0.485±0.378)g,方差分析表明用药组与对照组相比有显著性差异(t=5.59P<0.01),用药组抑瘤率达68.42%。结论:靶向Plk1的反义治疗能够抑制肿瘤的生长,对正常细胞无明显毒性。Plk1可作为肿瘤治疗的新靶点。Objective: To screen antitumor drugs targeted at Plk1 gene. Methods: Eight different 20 mer ASODN were designed according to the mRNA secondary structure of Plk1 gene and transfected into tumor cells by lipofection. Cell growth activities were estimated by MTT assay. One sequence was screened and optimized. The best sequence(No.9) with antitumor activity in vitro and vivo was analyzed. Results: Inhibition rates of No.5 targeted at sequences in the human Plk1 5′ un translateded mRNA on HepG2 cells were 59.1%, 75.5%, and 90.7% at the concentrations of 0.2 μmol/L,0.4 μmol/L, and 0.8 μmol/L, respectively. Its inhibiton rate was strongest. Inhibition rate of No.9 which removed 3 bases from 5′ un translated mRNA region of No.5 was strongest in four ASODNs. After No.9 transfected at the concentration of 0.2 μmol/L by lipofection, inhibition rate was increasing gradually, and attained its peak at the 4th day, than descended. It attained 78.3% at the 6th day. Nudes burdened HepG2 cells were adminstered No.9 at 25 mg/kg/day for 28 days. When they were killed, tumor weights of treated group were 0.485±0.378g. Control group were 1.536±0.196g. Student t test showed there was obvious difference between two groups. The inhibition rate of drug treated group was 68.42%. Conclusion: The results suggest that anti sense therapy specific to Plk1 mRNA is a practical approach to selective suppression of tumor growth, with little effect on normal cells.Plk1 could be used as an appropriate target for tumor therapay.

关 键 词：PLK1 反义寡核苷酸 HEPG2 肝癌 抗肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]