LncRNA ENST00000607314在胆囊癌化疗耐药中的作用及机制研究

Chemoresistance or chemotherapy resistance always limited the effects of comprehensive treatment on gallbladder carcinoma. The role of long non-coding RNA (LncRNA) in chemoresistance has been extensively explored, however, the studies that related to the chemoresistance of gallbladder carcinoma are rare. To identify transcripts that potentially drive the chemoresistance of gallbladder carcinoma, we determined the LncRNA expression profiles by microarray analysis and found that LncRNA ENST00000607314 is abundantly expressed in the chemoresistant gallbladder carcinoma cells. Along with other preliminary experiments results, we propose a hypothesis that ENST00000607314 may promote the chemoresistance of gallbladder carcinoma cells through recruiting FOXD1 to the promoter of ABCC2 and p27, and epigenetically regulating their transcript. In order to further explore the exact mechanism, this research will utilize a series of experimental methods to clarify the role of ENST00000607314 in the chemotherapy resistance of gallbladder carcinoma. Through targeted intervention ENST00000607314/FOXD1/ABCC2 and/or p27 pathway, we expected to identify the potential targets to reverse the chemotherapy resistance of gallbladder carcinoma in clinical practice.

化疗耐药是影响胆囊癌综合治疗效果的关键，LncRNA在耐药中的作用机制被广泛探索并报道，胆囊癌中相关研究结论尚不多见。本课题利用基因芯片技术发现LncRNA ENST00000607314在胆囊癌耐药细胞中呈高丰度表达并与耐药相关。根据前期实验结果，我们发现胆囊癌耐药细胞中ENST00000607314可与转录因子FOXD1结合并影响耐药相关蛋白ABCC2、p27表达，从而介导胆囊癌化疗耐药。为进一步探索其中的确切机制，本课题拟从分子、细胞、动物模型及临床标本等多层次并运用一系列实验方法以期明确ENST00000607314在胆囊癌化疗耐药中的作用，通过靶向干预ENST00000607314/FOXD1/ABCC2和/或p27通路逆转胆囊癌化疗耐药，为临床治疗胆囊癌挖掘潜在的治疗靶点。

胆囊癌（Gallbladder cancer，GBC）近年来发病呈增长趋势，大多数病人发现即为晚期，失去手术机会，化疗成为主要的治疗方式，然而现有一线化疗的总体有效率仅有30%左右，逆转化疗耐药是胆囊癌治疗疗效的关键。本课题组致力于胆囊癌化疗耐药的相关研究，在本项目中构建了胆囊癌吉西他滨化疗耐药的细胞株，并通过lincRNA芯片发现ENST00000607314 在GBC耐药细胞中表达升高，并且抑制ENST00000607314 的表达可以逆转化疗耐药；通过RNA pull-down实验检测到ENST00000607314与FOXD1的结合；随后利用ChIP检测到FOXD1与ABCC2、p27基因启动子的结合。除此之外，我们也在胆囊癌中证实光热疗法可以有效逆转化疗药物（Dox）的耐药。本研究旨在为克服胆囊癌耐药的基础和临床实验提供理论和方法依据，并为接下来深入的胆囊癌耐药机制的研究提供方向。