Yegorov Y E, Kazimirchuk E V, Terekhov S M, Karachentsev D N, Shirokova E A, Khandazhinskaya A L, Meshcheryakova J A, Corey D R, Zelenin A V
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov str. Moscow, 119991, Russia.
Cell Biol Int. 2002;26(12):1019-27. doi: 10.1006/cbir.2002.0961.
In previous work we demonstrated that various types of cultured cells with a limited life span could not reactivate DNA synthesis in the nuclei of mouse peritoneal macrophages in heterokaryons. We now investigate the role of telomerase in the process of the macrophage nucleus reactivation in heterokaryons with immortal telomerase-positive 3T3 Swiss mouse fibroblasts and human fibroblasts with introduced hTERT gene. We report that introduction of the hTERT gene into human diploid fibroblasts results in emergence of telomerase activity in these cells and the ability to induce the reactivation of DNA synthesis in the macrophage nuclei in heterokaryons. Inhibition of telomerase activity in heterokaryons by reverse transcriptase inhibitors (azidothymidine and guanosine polyphosphonate analogues) and by a 2'-O-methyl-RNA oligonucleotide anti-sense to the template region of telomerase RNA, block reactivation of DNA synthesis in macrophage nuclei without inhibiting DNA synthesis in the nuclei of fibroblasts. Our results suggest alterations (shortening or damage) in the macrophage telomere structure. As far as we know, heterokaryons with macrophages are the first cellular model for rapid investigation of the effects of telomerase inhibitors.
在之前的工作中，我们证明了多种寿命有限的培养细胞无法在异核体中的小鼠腹膜巨噬细胞核内重新激活DNA合成。我们现在利用永生化的端粒酶阳性3T3瑞士小鼠成纤维细胞和导入hTERT基因的人成纤维细胞，研究端粒酶在异核体中巨噬细胞核重新激活过程中的作用。我们报告称，将hTERT基因导入人二倍体成纤维细胞会导致这些细胞出现端粒酶活性，并具备在异核体中诱导巨噬细胞核内DNA合成重新激活的能力。用逆转录酶抑制剂（叠氮胸苷和鸟苷多磷酸酯类似物）以及针对端粒酶RNA模板区域的2'-O-甲基RNA反义寡核苷酸抑制异核体中的端粒酶活性，会阻断巨噬细胞核内DNA合成的重新激活，而不会抑制成纤维细胞核内的DNA合成。我们的结果表明巨噬细胞端粒结构发生了改变（缩短或损伤）。据我们所知，含有巨噬细胞的异核体是快速研究端粒酶抑制剂作用的首个细胞模型。
