MS2噬菌体病毒样颗粒包裹非编码RNA用于肝细胞癌诊断和治疗的研究

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide with poor prognosis and high lethality. Recently, studies have highlighted that dysregulated long non-coding RNAs (lncRNAs) in the tissue of HCC might play critical roles in HCC progression, which suggested that lncRNAs are emerging as novel biomarkers and potential targets in the treatment. As a kind of RNA, lncRNAs are easy to be degraded by RNase A. The concentration and quality of lncRNAs can be confounded by multiple technical factors in the analytical procedures. Therefore, normalization is an important aspect to measure lncRNAs, which is expected to reflect the expression level and responses to therapy of lncRNAs exactly. In this study, we decide to produce recombinant bacteriophage MS2 virus-like particles (MS2 VLPs) packaging lncRNAs as quality control materials in lncRNAs detection. Meanwhile, we produce MS2 VLPs packaging lncRNA-LET and shHULC with HCC specific cell-penetrating peptide on the surface by a yeast expression system. The therapy application of MS2 VLPs along with the changes of expression level of lncRNAs will also be analyzed. In this study, we will first obtain a new type of quality control materials for lncRNAs detection and establish a system in lncRNAs and siRNAs delivery.

肝细胞癌是世界范围内常见的恶性肿瘤，预后差、死亡率高。研究提示lncRNAs在肝细胞癌发生发展过程中发挥重要作用，并有望成为肝细胞癌的诊断标志物和治疗靶点。作为裸露的RNA分子，lncRNAs很容易被环境中的核糖核酸酶降解且其检测过程操作复杂易造成结果的偏差，因此临床研究中急需lncRNAs的质控品来监控检测过程，确保结果准确，以更好的评估临床标本中lncRNAs的表达情况及治疗前后的水平变化。本研究拟采用酿酒酵母真核表达技术制备包裹lncRNAs的重组病毒样颗粒作为监控lncRNAs检测过程的质控品；同时制备展示特异性短肽、包裹lncRNA-LET和siHULC前体的重组病毒样颗粒，探讨其介导的lncRNA-LET和lncRNA-HULC表达水平变化对肝细胞癌的治疗作用。本研究中我们将首次获得lncRNAs检测质控品并为lncRNAs和siRNAs同时特异性转运提供全新的通用载体。