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目的探讨hTERT基因的两端部分硫代修饰反义寡核苷酸(ASPS-ODN)对前列腺癌细胞PC3端粒酶活性的抑制作用。方法采用端粒重复序列扩增法及TRAP-PCR-ELISA法检测前列腺癌细胞的端粒酶活性;采用RT-PCR方法检测hTERT基因mRNA的表达水平;以免疫组化通过流式细胞仪检测hTERT基因蛋白水平的变化。结果hTERT基因的两端部分硫代修饰ASPS-ODN作用于PC3细胞48h,其端粒酶活性下降,作用72h,其端粒酶活性受到抑制。结论通过hTERT基因的两端部分硫代修饰ASPS-ODN特异性抑制hTERT基因mRNA的表达,可降低前列腺癌细胞PC3端粒酶活性。
Objective To investigate the inhibitory effect of ASPS-ODN on the telomerase activity of prostate cancer cell line PC3 by hTERT gene-containing antisense oligonucleotide. Methods The telomerase activity of prostate cancer cells was detected by telomeric repeat amplification and TRAP-PCR-ELISA. The expression of hTERT mRNA was detected by RT-PCR. The expression of hTERT mRNA was detected by flow cytometry Changes in gene protein levels. Results The telomerase activity of PC3 cells treated with ASPS-ODN at both ends of hTERT gene for 48 h decreased. The telomerase activity was inhibited after 72 h treatment. CONCLUSIONS: ASPS-ODN is a specific inhibitor of hTERT gene mRNA expression at both ends of hTERT gene, which can reduce the telomerase activity of prostate cancer PC3 cells.