Intronic miR-944联合Host gene p63在肺鳞癌中的作用机制及其诊断价值研究

Intronic miRNAs, which are generally resided within introns of the “host” genes, exhibit complex functional associations with the host genes in regulating several cancer-associated pathways. Exploring the putative interactions between them may not only illustrate the effects of intronic miRNAs in cancers, but also influence the interpretation of the action of host genes, which provides a novel strategy for revealing the etiology and pathogenesis of human cancers. Our previous data found that the expression level of miR-944 was significantly higher in lung squamous cell carcinomas (SCC) compared to adenocarcinomas (AC) by deep sequencing. Furthermore, we found that miR-944 was located in the intron of p63 host gene coincidently. However, the interaction between intronic miR-944 and p63 expression, the functional relationship of miR-944 and p63 in mediating tumorigenesis of lung cancer, as well as the possible value of miR-944 in distinguishing SCC from AC are largely unknown. Therefore, in the present study, we proposed to determine the effects between miR-944 and p63 at transcription and expression level by bio-software, in vitro test and clinical data; and to explore the effects and the possible mechanism of miR-944 cooperating with p63 in regulating the tumorigenesis and progression of lung cancer through in vitro, in vivo and clinical studies. Considering the critical role of host gene p63 in distinguishing SCC from AC, we next determined to clarify the value of miR-944 in lung cancer classification both in resected lung carcinomas and small biopsies/aspirates tumors by comparation of miRNA-based method and the standard pathologic methods of classification (i.e., microscopy and immunohistochemistry). Moreover, the universal significance of miR-944 in identifying SCC of different histological origin would be evaluated further. Our results may throw new lights on the role of miRNA-host gene cooperation in lung tumorigenesis, and identify specific miRNA biomarker of SCC to accurate subclassification of Non-small-cell lung cancer.

近期研究表明，Intronic miRNA可协同其定位基因（Host gene）参与肿瘤的发生发展过程，探究两者间相关性为揭示肿瘤发病机制提供了新方向。我们前期研究发现，Intronic miR-944与Host gene p63一样在肺鳞癌的表达显著高于腺癌。但目前有关miR-944与p63表达的相关性、单独或联合在肺鳞癌中的作用尚不清楚。本研究拟在前期工作基础上，探讨miR-944与p63在转录及表达间的调控作用；从癌变及进展过程揭示miR-944联合p63在肺鳞癌中的作用机制。此外，鉴于p63在肺癌鉴别诊断中的意义，拟进一步利用手术切除及穿刺组织明确miR-944在肺鳞癌与腺癌诊断中的价值；评价miR-944在不同来源鳞癌诊断中的作用。研究旨在揭示Intronic miR-944—p63相互作用在肺鳞癌中的作用机制，发现与肺鳞癌特异相关miRNA，为肺癌的准确诊断及有效治疗奠定基础。

肺癌是世界范围内最高发的恶性肿瘤之一，深入探讨其发病机制、准确鉴别肺鳞癌与腺癌，对于非小细胞肺癌患者的诊断和治疗意义重大。MicroRNA以其高度的稳定性突显了其作为生物标记分子的重要意义。我们前期研究发现，Intronic miR-944与Host gene p63一样在肺鳞癌的表达显著高于腺癌。但是，有关miR-944在肺鳞癌与腺癌鉴别诊断中的价值、miR-944与其Host gene p63表达的相关性等问题尚不明确。因此，本研究主要从：miRNAs（miR-944、miR-205-5p）在肺鳞癌与肺腺癌鉴别诊断中的意义；miR-944与其Host gene p63基因间相互的调控作用；miR-944与其p63表达间反馈调节机制的探讨；肺鳞癌与腺癌组织中差异基因表达的筛选四方面进行了研究。研究结果如下：1）建立了基于miRNAs（miR-944与miR-205-5p）的肺鳞癌与肺腺癌鉴别诊断模型，尤其在支气管灌洗液中miRNAs检测对肺癌诊断及病理分型鉴别诊断的研究结果，为肺癌患者个体化治疗方案的制定提供高灵敏度和特异性的生物标记，具有重要的临床转化潜能。2）p63为目前临床工作中肺鳞癌特异性标记分子，而miR-944为p63基因内含子区miRNA，两者以正反馈方式相互调节表达，体现了Intronic miRNA与Host gene基因间复杂的调控作用，也更加证实了miR-944作为肺鳞癌标记分子的意义。3）YAP1/TAZ可抑制细胞中p63基因表达，而miR-944过表达可抑制YAP1或TAZ表达，解除YAP1/TAZ对p63基因的抑制作用，是miR-944促进其Host gene p63表达的机制之一。本项目的实施为后续研究开启了新的研究方向，对于揭示非小细胞肺癌发生发展的病因及机制研究奠定了重要基础。