Regulatory RNAs and the design of cancer therapy using RNA biotechnology

近年来，非编码rna（调控rna）在基因表达调控中发挥着重要作用。本研究旨在阐明microRNAs在造血分化和白血病发生中的作用，阐明端粒酶活性的调控机制，并在调控RNA生理功能的基础上，利用RNA生物技术开发癌症诊断和治疗的新方法。得到以下结果：i) sirna敲低白血病融合基因，导致自我更新活性下调，诱导分化，提示这些sirna可以作为白血病的治疗药物；ii) HL-60细胞向单核/巨噬细胞分化过程中MicroRNA表达动态变化；iii)获得了抗KRAS癌蛋白和其他癌症相关蛋白的RNA适配体，这将为癌症诊断和治疗的新方法奠定基础；端粒酶是一种特殊的逆转录酶，它利用固有的RNA模板合成端粒DNA。虽然已经确定了编码催化亚基和模板RNA的基因，但对体内端粒酶如何被募集到端粒知之甚少。在本研究中，我们试图在人类和裂变酵母中进行生化鉴定。我们发现人类端粒酶对提取有很强的抵抗力。然而，我们分离出了分裂酵母蛋白Poz1和Ccq1，它们负向和正向调节端粒酶。

It has recently been revealed that non-coding RNAs (regulatory RNAs) play important roles in the regulation of gene expression. The aims of this study were to manifest the role of microRNAs in hematopoietic differentiation and leukemogenesis, to clarify the regulation of telomerase activity, and to develop new methods for cancer diagnosis and therapy using RNA biotechnology on the basis of the physiological functions of regulatory RNAs. The following results were obtained: i) Knockdown of the leukemia fusion gene by siRNAs resulted in down-regulation of self-renewal activity and induction of differentiation, suggesting that these siRNAs can be used as therapeutics for leukemia; ii) MicroRNA expressions changed dynamically during differentiation to monocyte/macrophage in HL-60 cells; iii) RNA aptamers against the KRAS oncoprotein and other cancer-related proteins were obtained, which will become seeds for new methods of cancer diagnosis and therapy; iv) Telomerase is a specialized reverse transcriptase that synthesizes telomeric DNA using an intrinsic RNA template. Although the genes encoding the catalytic subunit and the template RNA have been identified, little is known about how telomerase is recruited to telomeres in vivo. In this study, we tried to biochemically identify such factors in human and fission yeast. We found that human telomerase is very resistant to extraction. However, we isolated fission yeast proteins Poz1 and Ccq1 that negatively and positively regulate telomerase.