lncRNA-ATB对TGF-β诱导的胃癌上皮间质转化的调控作用及消痰散结方干预机制研究

The epithelial-mesenchymal transition(EMT) play a critical role in invasion-metastasis cascade of tumor, long noncoding RNA(lncRNA) has been reported to mediate the EMT in tumor progression. Recent study found that the lncRNA activated by TGF-β(lncRNA-ATB) induced EMT and invasion by competitively binding the miR-200 family and IL-11 mRNA in hepatocellular carcinoma and breast cancer, but its mechanism in gastric cancer(GC) is not fully understood. According to our previous research, Xiaotan Sanjie Formula(XTSJF) could prolong the survival time in GC patients, decrease the concentration of TGF-β and IL-8, inhibit fibroblast activation protein(FAP) expression in experiment study. Accordingly, we presume that XTSJF might inhibit EMT through mediate the lncRNA-ATB in GC metastasis. In present study, we intend to explore the mechanism of the lncRNA-ATB mediate EMT in GC lines, and XTSJF whether or not interfere lncRNA-ATB exprssion to inhabit EMT in GC in vitro and in vivo used western blot、quantitative reverse transcriptase polymerase chain reaction(qRT-PCR) methods. This will provide the experiment basis for the early recognition EMT, and give new insight toward therapeutic intervention against of GC, and provide scientific basis for the TCM and Chinese herbal in gastric cancer treatment.

上皮间质转化(EMT)是肿瘤侵袭转移级联过程的关键步骤，长链非编码RNA（lncRNA）参与了EMT进程。最近研究表明，转化生长因子激活的RNA(lncRNA-ATB)通过竞争性结合miR-200s和IL-11参与EMT形成，但是其在TGF-B诱导的胃癌EMT中的作用尚不完全清楚。前期研究表明消痰散结方能延长胃癌患者生存期，减少原位移植胃癌裸鼠血清IL-8、TGF-β炎症相关因子的浓度、减少FAP蛋白表达的作用，可能是其抑制胃癌侵袭转移的机制之一，因此我们推测消痰散结方可能通过调控lncRNA-ATB抑制胃癌EMT，以达到抑制侵袭转移之目的。本项目通过构建lncRNA-ATB过表达/干扰载体，采用western blot、qRT-PCR等方法，研究lncRNA-ATB参与胃癌EMT过程及调控机制，以及消痰散结方对其干预机制，为抑制胃癌转移提供理论支持，为中西医结合胃癌治疗提供科学依据。

晚期胃癌转移的5年生存率低于20%，上皮间质转化上皮间质转化(EMT)是肿瘤侵袭转移级联过程的关键步骤，转化生长因子激活的RNA(lncRNA-ATB)通过竞争性结合miR-200s和IL-11参与EMT形成。基于从“痰论治理论”的消痰散结方在体内体外都具有明显抑制胃癌侵袭转移。但消痰散结方是否通过调控lncRNA-ATB从而抑制TGF-β诱导的胃癌EMT的机制尚不明确。本项目通过对多种胃癌细胞lncRNA-ATB表达水平的检测、lncRNA-ATB过表达和干扰、TGF-β诱导胃癌EMT和干细胞化等细胞分子学生物手段证明lncRNA-ATB在不同的胃癌细胞中的作用及受TGF-β的诱导存在差异。消痰散结方对不同胃癌细胞的抑制效果也不同，但对高表达lncRNA-ATB的HGC-27和MNK-45细胞具有更好的抑制效果，抑制lncRNA-ATB、EMT和细胞周期等相关蛋白的表达，同时miR-200的表达上调导致ZEB等转录因子下调。小鼠体内实验发现：消痰散结方能够显著抑制胃癌原位移植瘤生长和TGF-β、IL-18等转移相关细胞因子的表达。这些结果进一步证明了中医“从痰论治理”论指导下的消痰散结方临床疗效的真实性和潜在的治疗靶向及分子标志物，本项目为消痰散结方的后期开发和临床应用提供重要的研究基础。