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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 the effect of ASODN on the sensitivity of glioma U251 cells to mitomycin C (MMC). Methods The c-Met mRNA of U251 cells was blocked with 5 μmol / L c-Met ASODN, 50 μg / L MMC was cocultured with it, and the expression of c-Met mRNA was detected by reverse transcription-polymerase chain reaction The growth of U251 cells was detected by MTT assay, the expression of PCNA protein was detected by immunohistochemistry and the cell adhesion rate was detected in vitro. Non-sense oligonucleotide-treated and untreated U251 cells served as controls. Results The expression of c-Met mRNA in U251 cells treated with c-Met ASODN (absorbance value of 62 ± 21) was significantly lower than that of U251 cells treated with non-sense oligonucleotide (absorbance value of 150 ± 25, P <0. (53.84 ± 12.21)%, (14.61 ± 3.82)% and (0.35 ± 0), respectively. The sensitivity to MMC, cell growth inhibition rate, cell adhesion rate and PCNA index were .02)%, significantly higher than the corresponding nonsense [(9.86 ± 3.42)%, (24.84 ± 5.90)% and (0.55 ± 0.04)%, P <0. 05]. Conclusion c-Met ASODN can enhance the sensitivity of glioma cell line U251 to MMC. The molecular mechanism may be related to the downregulation of c-Met gene and PCNA protein by c-Met antisense oligonucleotide.