miR214对肺腺癌干细胞自我更新的调控作用及机制

Self-renewal is the most characteristic of lung adenocarcinoma cancer stem cells (LASCs) and miRNAs are involved in self-renewal of LASCs. In our primary experiments, we found exciting results as follows: miR214 is unregulated in LASCs compared to cancer cells and knock-down miR214 could reduce sphere formatin of LASCs; there is negative colleration between miR214 and MITF protein expression. Based on the previous results, we propose that miR214 is involved in LASCs regulation by inbibiting MITF expression. We would use lentivirus-mediated gene trasfer to overexpress or knock down miR214 to determine the role of miR214 in LASCs self-renewal in vitro and in vivo. Furthermore, we would elucidate the underlying mechanisms of miR-214-regulating LASCs self-renewal by co-transfecting experiments. Also, we would validate the results by samples collected from clinic patients with lung adenocarcinoma by analyzing the correlation of LASCs percentage and miR214/MITF expression. The present project will help to elucidate the underlying mechanisms of LASCs self-renewal, but also provide a new avenue for LASCs targeting therapy.

自我更新是肺腺癌干细胞（LASCs）最重要的生物学特征；miRNAs在肿瘤干细胞的自我更新中发挥重要作用。本项目组前期研究发现：1.LASCs高表达miR214，抑制miR214能降低LASCs体外成球能力；2.miR214与MITF蛋白表达呈负相关。我们提出"miR214通过抑制MITF表达调控LASCs自我更新"的学术假说。拟在前期实验基础上，采用慢病毒介导的miR214/miR214 inhibitor转染LASCs并移植裸鼠体内，明确miR214在LASCs自我更新中的作用；采用共转染等技术验证miR214抑制MITF，阐明miR214对LASCs自我更新影响的机制；收集临床标本，通过检测miR214和MITF表达，同时进行LASCs比例分析，在临床肺腺癌的患者中进一步验证学术假说。通过本项目的研究不但能完善LASCs自我更新的分子机制，而且也为肺腺癌干细胞靶向治疗提供科学依据。

自我更新是肺腺癌干细胞(LASCs) 最重要的生物学特征;miRNAs在肿瘤干细胞的自我更新中发挥重要作用.本项目组研究发现:1.A549细胞株与原代组织来源的LASCs高表达miR214,抑制miR214能降低LASCs体外成球能力;2.应用在线靶基因预测软件Targetscan(http://www.targetscan.org),Pictar(http://pictar.mdc-berlin.de)和microRNA.org (http://www.microrna.org)对miR-214的靶基因进行预测，先找出三个软件预测到的miR214的靶基因交集，并通过GO（Gene Ontology）注释对靶基因的生物学功能进行分析，找出与自我更新相关的靶基因。3.miR-214 靶基因的验证。