LINC01060调控RhoA-ROCK信号通路对胰腺癌侵袭转移的影响及机制研究

Making the mechanism of invasion and metastasis of pancreatic cancer certain is very important to improve its treatment status. LncRNAs play a vital role in the invasion and metastasis process of a wide variety of tumors, but the relationship between LncRNAs and pancreatic cancer is not deeply studied. According to early lncRNA microarray screening and expanded validation experiments, we found that LINC01060 overexpressed in pancreatic cancer tissues compared with adjacent no-tumor tissues, and the level of LINC01060 was related with the lymph node and distant metastasis status. Silencing LINC01060 in the experiments inhibited invasion and metastasis of pancreatic cancer in vivo and in vitro, and down-regulated RhoA, ROCK1 and ROCK2 expression. In our earlier study we proved that RhoA-ROCK signaling pathways play an important role in the process of invasion and metastasis of pancreatic cancer. As a result we hypothesis that LINC01060 regulates RhoA-ROCK signaling pathways to enhance the invasion and metastasis of pancreatic cancer. We will test the LINC01060 level in an expanded sample size and analysis its correlation with the invasion and metastasis of pancreatic cancer. And we will clarify its influence on the invasion and metastasis of pancreatic cancer in vivo and in vitro. We will illustrate the mechanism underlying which LINC01060 regulates RhoA - ROCK signaling pathways through RIP, CLIP experiments. This study is expected to further reveal the mechanism of invasion and metastasis of pancreatic cancer and provide a new therapeutic target of pancreatic cancer.

明确胰腺癌侵袭转移的机制对改善其治疗现状非常必要，LncRNA在多种肿瘤侵袭转移过程中发挥关键作用，但lncRNA与胰腺癌侵袭转移的关系还没有深入的研究。我们前期芯片筛选并扩大样本量检测发现LINC01060在胰腺癌中表达量与淋巴结和远处转移正相关；沉默LINC01060在体内外实验中均抑制胰腺癌的侵袭转移，并下调RhoA、ROCK1和ROCK2表达。我们既往研究表明RhoA-ROCK信号通路在胰腺癌侵袭转移过程中发挥关键作用。所以提出LINC01060调控RhoA-ROCK信号通路增强胰腺癌侵袭转移的假设。拟扩大样本量检测LINC01060并分析与胰腺癌转移的相关性；体内外实验研究其对胰腺癌侵袭转移的影响；通过RIP、CLIP等实验阐明LINC01060调控RhoA-ROCK信号通路的机制。本研究有望进一步揭示胰腺侵袭转移的机制，为胰腺癌治疗提供新的靶点。

胰腺癌侵袭转移的机制仍然是当前的研究热点和难点，在很多肿瘤中lncRNA都在侵袭转移过程中发挥关键作用，但lncRNA与胰腺癌侵袭转移的关系还没有进行深入的研究。我们旨在鉴定在胰腺癌进展中具有重要作用的lncRNA。首先我们筛选了在胰腺癌组织中差异表达的lncRNA。在349个差异表达的lncRNA中，与正常胰腺组织相比，Linc01060在胰腺癌组织中的表达最低。胰腺癌组织中Linc01060的低表达与不良预后显着相关。Linc01060在体外和体内抑制胰腺癌的增殖和侵袭。Vinculin的过表达抑制了Linc01060KD介导的FAK和paxillin磷酸化的增加，而Vinculin敲低逆转了Linc01060介导的FAK的抑制和粘着斑转换的失活。 Vinculin敲低还可以通过上调ERK活性来促进胰腺癌细胞的增殖。在生物学功能分析中，蛋白Vinculin的过度表达消除了Linc01060介导的胰腺癌细胞增殖和侵袭的抑制，而蛋白Vinculin抵消了Linc01060介导的PC细胞增殖和侵袭的抑制。这些数据表明，Linc01060在通过调节Vinculin表达来抑制胰腺癌进展中起关键作用。这些发现表明，Linc01060-vinculin-焦点黏附轴是胰腺癌治疗的治疗靶标。