长链非编码RNA LINC00326在5-Aza介导的非小细胞肺癌转移抑制中的机制研究

The role and mechanism of methylated transferase inhibitors in lung cancer metastasis are not yet clear. Our previous studies have found that 5 -Aza can restrain high metastatic non-small cell lung cancer cell migration and invasion and regulate EMT/MET. Furthermore, we screened and identified lncRNA LINC00326 that was differently expressed in lung cancer under the treatment with 5-Aza. We think that lncRNA LINC00326 is involved in regulation of lung cancer cell EMT /MET balance and inhibition of lung cancer metastasis by 5-Aza, and hypothesize that, by means of lncRNA LINC00326 overexpression, 5-Aza can inhibit lung cancer metastasis by regulating PDCD1-PI3K/AKT signal pathway mediated by miR-657. This study aims to explore the mechanism of 5-Aza in the inhibition of lung cancer metastasis by studying lncRNA LIN00326 expression inhibited by 5-Aza, and regulation of PDCD10-PI3K/AKT signal pathway mediated by miR-657. It will not only provide new targets for the treatment of non-small cell lung cancer, but also provide new ideas and directions for epigenetic combined immunotherapy in non small cell lung cancer.

甲基化转移酶抑制剂在肺癌转移方面的作用和机制尚未明确。我们前期研究发现5-Aza能抑制高转移性非小细胞肺癌细胞的迁移和侵袭能力，并调控EMT/MET平衡，在此基础上筛选出5-Aza处理前后差异表达的lncRNA LINC00326，推测其参与了5-Aza调控肺癌细胞EMT/MET平衡，提出“5-Aza 通过上调lncRNA LINC00326，调节miR-657介导的PDCD1-PI3K/AKT信号通路抑制非小细胞肺癌转移”。本课题通过检测5-Aza上调lncRNA LINC00326表达，调节上述信号通路以论证该假说，探究5-Aza抑制肺癌转移具体机制，为非小细胞肺癌治疗提供新的靶点，也为表观遗传联合免疫治疗非小细胞肺癌提供新的思路和方向。

非小细胞肺癌（non-small cell lung cancer, NSCLC）是一种常见又具备高度侵袭性的恶性肿瘤，尽管它的发生机制和治疗方法已有大量的研究，有效的干预手段仍然很少。项目组前期研究发现表观遗传治疗药物氮杂胞苷（5-Aza）干预可抑制NSCLC细胞迁移和侵袭。在5-Aza干预后差异表达的lncRNAs中，我们筛选到lncRNA LINC00326，生信预测LINC00326靶向调控miR-657。我们推测5-Aza上调LINC00326表达，打破上皮间质转化平衡，抑制NSCLC迁移侵袭。该课题主要目标是，确认5-Aza调控的LINC00326靶向miR-657参与NSCLC生长和转移的生物学行为。在课题执行中，我们发现在细胞和动物水平，5-Aza均可抑制肺癌生长。临床样本和细胞中均测试到NSCLC的miR-657高表达，LINC00326低表达，dickkopf Wnt信号通路抑制剂2（dickkopf Wnt signaling pathway inhibitor 2, DKK2）低表达，SRC激酶信号抑制剂1（SRC kinase signaling inhibitor 1, SRCN1）低表达。通过一系列实验验证LINC00326靶向调控miR-657，miR-657靶向调控DKK2和SRCIN1。确定LINC00326调控miR-657靶向DKK2参与调控Wnt/β-catenin信号通路，同时miR-657靶向SRCIN1参与NSCLC上皮间质转化。基于此，假说基本得到验证。本研究从细胞-分子-动物-临床四个层面，系统性的阐述了LINC00326介导的miR-657调控Wnt/β-catenin信号通路和肿瘤细胞上皮间质转化的作用机制，为5-Aza在NSCLC转移抑制方面提供了新的证据，也为NSCLC的治疗新靶点研究提供了理论性实验基础。