硫代修饰型反义寡核苷酸抑制肺癌细胞系端粒酶活性及增殖的研究

了解人肺癌细胞系的端粒酶活性情况。探讨硫代修饰型端粒酶ＲＮＡ反义寡核苷酸封阻端粒酶ＲＮＡ,对癌细胞代谢,生长的抑制作用。方法采用端粒酶ＰＣＲ ＥＬＩＳＡ方法检测８种体外培养从肺癌细胞系端粒酶活性。合成端粒酶ＲＮＡ的反义硫代寡核苷酸（６聚体、９聚体、１２聚体）和随机序列寡核苷酸,分别导入ＬＴＥＰ－ａ－２细胞系,作用７２ｈ。采用端粒酶ＰＣＲ ＥＬＩＳＡ方法检测端粒酶活性,四甲基偶氮唑盐（ＭＴＴ）光吸收

Inhibition of hTR activity for suppression of lung cancer cell proliferation using antisense phosphorothioate oligonucleotides

OBJECTIVE: Telomerase activity was measured in 8 lung tumor cell lines and transfected antisense phosphorothiate oligodeoxynucleotides (anti-PS-ODN) of hTR as therapeutic agents into target cell LTEP-a-2 to investigate the inhibitory effect on telomerase activity and tumor cell growth. METHODS: 1. Telomerase activity assay in 8 human lung tumor cell lines using telomerase PCR ELISA. 2. Synthesized anti-PS-ODNs of hTR and random PS-ODN transfected with Clonfectin into LTEP-a-2 cell lines for 72 hr. 3. Measure telomerase activity by telomerase PCR ELISA, SDH activity by MTT assay and cell growth. RESULTS: Eight cell lines showed positive expression of telomerase activity. Various anti-PS-ODNs could inhibit telomerase activity, SDH activity and cell growth. The inhibition became more marked with the increase of anti-PS-ODNs concentrations. Concentrations of 5 - 40 micromol/L anti-PS-ODNs of hTR specifically reduced the telomerase activity by 25.7% - 84.0%. PS-ODNs of anti-hTR were also able to reduce SDH activity by 19.4% - 74.7% at 5 - 40 micromol/L. The dose of 5 - 40 micromol/L PS-ODNs of anti-hTR had the ability to inhibit cell growth by 14.8% - 72.5%. The results indicate that random sequence of PS-ODN(9) failed to inhibit telomerase, SDH activity and cell growth. A statistically significant difference exists between random PS-ODN and three anti-PS-ODNs (P < 0.01). CONCLUSIONS: Human lung tumor cell lines express high telomerase activity. PS-ODNs of anti-hTR had the ability to inhibit telomerase and reduce LTEP-a-2 cell growth and metabolism.