长链非编码RNA-NCRNA00173参与小细胞肺癌耐药的机制研究

我们前期发现长链非编码RNA-NCRNA00173与小细胞肺癌耐药关系密切，并可能作为一种竞争性内源RNA，参与小细胞肺癌耐药的调节。为进一步明确NCRNA00173在小细胞肺癌耐药中的作用及机制，本课题拟利用已有耐药细胞模型开展以下研究：（1）体外和体内实验分析NCRNA00173对细胞药物敏感性的影响；（2）荧光素酶实验验证miR-495/miR-218靶向作用NCRNA00173和Etk/BMX；（3）下调或上调NCRNA00173表达，同时增加或降低miR-495/miR-218，分析对Etk/BMX表达的影响；（4）RIP实验明确NCRNA00173通过miR-495/miR-218调节Etk/BMX表达；（5）分析NCRNA00173在小细胞肺癌组织中表达的临床意义及与miR-495/miR-218及Etk/BMX的相关性。通过本项目研究，进一步丰富小细胞肺癌耐药的分子机制。

Our previous study has demonstrated that long non-coding RNA (lncRNA) NCRNA00173 is closely associated with the chemoresistance of small cell lung cancer (SCLC). NCRNA00173 may function as competing endogenous RNAs (ceRNAs) in modulating SCLC drug resistance. To further investigate the molecular mechanism of NCRNA00173 in regulating chemoresistance of SCLC, we intends to use two different cell models of SCLC resistant cells to carry out the following studies: (1) To clarify the effect of NCRNA00173 on cell sensitivity to chemotherapeutic drugs in vitro and in vivo; (2) Using luciferase experiment to verify the miR-495/miR-218 can directly target NCRNA00173 and Etk/Bmx; (3) To analyze the effect of both miR-495/miR-218 and NCRNA00173 on Etk/Bmx expression; (4) Using RIP technology to analyze the role of NCRNA00173 on Etk/Bmx expression through miR-495/miR-218; (5) To analyze the effect of miR-495 methylation status on LINC00173 expression; (5) To analyze the clinical significance of NCRNA00173 expression and its relationsip with miR-495/miR-218 and Etk/Bmx in SCLC tissues. The molecular mechanism of SCLC chemoresistance will be further enriched through this research project.