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目的 有效抑制肿瘤细胞端粒酶活性 ,为肿瘤的基因治疗提供依据。方法 设计合成了针对端粒酶逆转录酶 m RNA5′端区的锤头状核酶 (5′- end of human telomerase reverse trascriptase m RNA,h TERT- 5′RZ)基因 ,并将该基因重组入 pc DNA3.1(+) ,经体外转录得到小片段 h TERT- 5′RZ。用脂质体介导法 ,将 h TERT- 5′RZ导入 He L a细胞以检测其对 He L a细胞端粒酶活性的抑制效力。同时比较了同法合成的针对端粒酶 RNA模板的核酶 (telomerase m RNA,telo RZ)和两种核酶混合物对 He L a细胞端粒酶活性的抑制效力。结果 小片段 h TERT- 5′RZ能有效抑制 He L a细胞端粒酶活性 ,且其抑制效力较单纯telo RZ为高 ,两种核酶混合物的端粒酶抑制效力与 h TERT- 5′RZ相当 ,也高于单纯 telo RZ。结论 小片段核酶 h TERT- 5′RZ可望成为优于 telo RZ的端粒酶抑制剂 ,在肿瘤基因治疗中发挥作用。
Objective Effective inhibition of telomerase activity in tumor cells provides the basis for gene therapy of tumors. Methods The 5’-end of human telomerase reverse trascriptase m RNA (h TERT-5’RZ) gene was designed and synthesized, and the gene was recombined into pcDNA3.1 (+), was obtained by in vitro transcription of small fragments h TERT- 5’RZ. H TERT-5’RZ was introduced into HeLa cells by liposome-mediated method to examine its inhibitory effect on the telomerase activity of HeLa cells. At the same time, telomerase RNA (telo RZ) and the mixture of two ribozymes synthesized by the same method were compared to inhibit the telomerase activity of HeLa cells. Results h TERT-5’RZ could effectively inhibit the telomerase activity of HeLa cells and its inhibitory potency was higher than that of telo RZ alone. The telomerase inhibitory potency of the two ribozyme mixtures was similar to that of hTERT-5’RZ Pretty much, but also higher than pure telo RZ. Conclusion A small fragment of ribozyme h TERT-5’RZ is expected to be a telomerase inhibitor superior to telo RZ and plays a role in tumor gene therapy.