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目的探讨hTERT基因的两端各三个碱基硫代修饰反义寡核苷酸(AS PS-ODN)对氟他胺耐受性前列腺癌细胞LNCaP端粒酶活性的影响机制。方法采用TRAP-PCR-ELISA法检测氟他胺耐受性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基因的两端各三个碱基硫代修饰反义寡核苷酸(AS PS-ODN)作用于LNCaP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基因的两端各三个碱基硫代修饰反义寡核苷酸(AS PS-ODN)特异性抑制hTERT基因mRNA的表达降低氟他胺耐受性前列腺癌细胞LNCaP端粒酶活性,为氟他胺耐受性前列腺肿瘤的基因治疗提供新思路。
OBJECTIVE: To investigate the effects of three basic thio-modified antisense oligonucleotides (hpsT) on the telomerase activity of flutamide-tolerant prostate cancer cells (LNCaP). Methods The telomerase activity of flutamide tolerant prostate cancer cells was detected by TRAP-PCR-ELISA. The expression of hTERT gene mRNA was detected by RT-PCR. The expression of hTERT gene protein was detected by flow cytometry Horizontal changes. Results The telomerase activity of LNCaP cells was down-regulated by three PS-ODN antisense oligodeoxynucleotides (AS PS-ODNs) at each end of hTERT gene for 48 h. The telomerase activity was inhibited after 72 h treatment. Conclusions The specific inhibition of hTERT gene mRNA expression by three PS-ODN antisense oligonucleotides at both ends of hTERT gene reduces the telomerase activity of LNCaP in flutamide tolerant prostate cancer cells , Which provides new ideas for the gene therapy of flutamide tolerant prostate tumor.