c-myc反义寡核苷酸对人肝癌细胞SMMC-7721端粒酶活性的影响

Effect of Antisense c-myc Oligonucleotide on Telomerase Activity in Human SMMC-7721 Liver Cancer Cells

目的 :探讨c -myc反义基因治疗肿瘤的机制。方法:将正义 (sense)、反义 (antisense)、错配 (mismatch)的c -myc寡核苷酸作用于体外培养的人肝癌细胞SMMC -7721 ,分别应用免疫组化、RT -PCR、PCR -ELISA等方法 ,观察对c -myc蛋白表达、端粒酶亚单位及活性表达的影响。结果:10μmol/L浓度的反义c -myc寡核苷酸作用于SMMC -772124h、48h后 ,相应的c -myc蛋白、端粒酶催化亚单位hTERTmRNA及其端粒酶活性与空白对照组相比 ,差别有统计学意义(P<0.01);而正义组和错配组与空白对照组相比 ,差别无统计学意义(P>0.05)。结论:c -myc反义寡核苷酸可能通过抑制端粒酶活性而抑制肿瘤细胞增殖。

Objective:To study the anti-cancer mechanism of antisense c-myc oligonucleotide.Methods:The sense,antisense and mismatch oligomers for c-myc were individually delivered to cultured SMMC-7721.Using immunohistochemistry,RT-PCR and PCR-ELISA,the effects on c-myc protein expression,telomerase-related major subunits hTR,hTERT,TP1mRNA and telomerase activity were observed.Results:Antisense c-myc,at the concentration of10μmol/L,could successfully inhibit the expression of c-myc proˉtein,hTERT mRNA and telomerase activity in SMMC-7721(P<0.01),but the sense and mismatch did not have these effects(P>0.05).Conclusion:The anti-cancer mechanism of antisense c-myc oligonucleotide may by its inhibitory effect on telomerase activity.