摘要
目的探讨肝细胞生长因子受体(c-Met)反义寡核苷酸(ASODN)增强胶质瘤细胞系 U251细胞对丝裂霉素C(MMC)敏感性的作用。方法用5 μmol／L c-Met ASODN封闭U251细胞 c-Met mRNA,将50μg/L的MMC与其共培养,采用逆转录-聚合酶链反应(RT-PCR)技术检测c- Met mRNA表达,噻唑蓝(MTT)试验检测U251细胞的生长情况,免疫组织化学法检测细胞PCNA 蛋白的表达,体外检测细胞黏附率。以无义寡核苷酸处理及未处理U251细胞为对照。结果经 c-Met ASODN处理的U251细胞 ,c-Met mRNA的表达(吸光度值为62±21)明显低于经无义寡核苷酸处理U251细胞(吸光度值为150±25,P<0．05);对MMC的敏感性,细胞生长抑制率、细胞黏附率及PCNA指数分别为(53．84±12．21)％、(14．61±3．82)％和(0．35±0．02)％,明显高于相应的无义[(9．86±3．42)％、(24．84±5．90)％和(0．55±0．04)％,P<0．05]。结论 c-Met ASODN能增强胶质瘤细胞系U251细胞对MMC的敏感性,其分子机制可能与c-Met反义寡核苷酸下调了c- Met基因和PCNA蛋白的表达有关。
Objective To investigate whether the c-Met antisense oligodeoxynudeotide （A- SODN） enhances the sensitivity of glioma cell U251 to mitomycin C. Methods The c-Met mRNA of 13251 cells was blocked by 5 μmol/L c-Met ASODN, the expression of c-Met mRNA was determined by RT-PCR, and its effect on cell growth was evaluated by methyl thiazolyl tetrazodium （MTT）, the expression of PCNA protein was detected by immunohistochemistry, and tumor cell adhesion in vitro measured. Untreated group and nonsense oligonucleotide group were regarded as control groups. Results The c-Met mRNA expression in c-Met antisense treated group were lower than in the nonsense treated group （P〈 0.05）. The sensitivity of 13251 to mitomycin C was increased in antisense treated group as compared with the nonsense treated group （ P 〈 0.05）. Conclusion The sensitivity of glioma cell 13251 to mitomycin C can be enhanced hy the blockage of the c-Met mRNA and PCNA protein expression.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第5期540-542,共3页
Chinese Journal of Experimental Surgery
基金
湖北省卫生厅科研基金资助项目(JX1B019)
