miR-194/GRHL2通路调控肺癌细胞增殖和转移的机制及临床意义研究

Lung cancer is the one of common malignant tumor in china, ranking first morbidity. Our previous study demonstrated that GHRL2 is considered to play an important regulatory role in the pathogenic process of tumors, but it is still not known what the function and regulation mechanism is in lung cancer. Recently, pre-experiment showed that GRHL2 was up-regulated expressed as an indicator of poor prognosis in lung cancer, and GRHL2 had good effect on cell proliferation, migration and invasion of lung carcinoma, the tumor suppressor gene miR-194 acted directly on the 3'-UTR region of GRHL2 to decrease its expression, which may preliminarily explain why GRHL2 was up-regulated expressed in lung carcinoma. Those results suggested that the down-regulated expression of miR-194 leads to the up-regulated expression of GRHL2 and the promotion of proliferation and metastasis of tumors. For validating our scientific hypothesis, we intend to further by lentiviral transduction, cell proliferation assay, animal experiment, gene chip, ChIP, dual luciferase reporter assay system manifest miR-194 promote lung cancer cell proliferation and metastasis by deregulating grhl2, and elucidate grhl2 function in promoting the molecular mechanisms of cancer. This project can be carried out to provide fresh ideas as new therapeutic targets based on miR-194 / GRHL2 pathways in lung cancer.

肺癌是发病率最高的恶性肿瘤,揭示肺癌的发病机理尤为迫切。GRHL2被认为在肿瘤的致病过程中发挥着重要的调控作用，但在肺癌中的功能及调控机理仍不明确。我们的前期研究表明：①GRHL2在肺癌中表达上调；②过表达GRHL2发现其具有促进肺癌细胞增殖、迁移和侵袭的作用；③抑癌基因miR-194作用于GRHL2的3’-UTR区域进而可负调控GRHL2基因的表达。由此我们提出科学假设：miR-194表达下调导致GRHL2表达上调并促进肺癌细胞的增殖和转移。为了验证本科学假设，我们拟进一步通过慢病毒转导技术、细胞增殖实验、动物实验、基因芯片、ChIP、双荧光素酶报告实验等以获取miR-194负调控GRHL2促进肺癌细胞增殖和转移的可靠证据，同时将阐明GRHL2发挥促癌作用的分子机理。本项目的开展为确立miR-194/GRHL2通路作为肺癌新的治疗靶点提供新的策略。

转录因子Grainyhead-like 2 (GRHL2)参与多种生物过程中的伤口愈合、表皮完整性和上皮-间充质转化(EMT)；然而，GRHL2在非小细胞肺癌中的生物学功能尚不清楚。在本项目中，我们研究了GRHL2对非小细胞肺癌细胞系和临床组织中细胞生长和迁移的影响。临床非小细胞肺癌标本的免疫组织化学分析显示，GRHL2高表达的患者与GRHL2低表达的患者相比，预后较差。GRHL2过表达促进细胞生长和集落形成，同时抑制非小细胞肺癌细胞的细胞迁移。此外，GRHL2通过直接与RhoG启动子区结合而降低了RhoG的转录活性。这些发现证实GRHL2在调节非小细胞肺癌细胞增殖和迁移中起重要作用。