外泌体lncRNA ANCR在骨肉瘤阿霉素耐药中的作用及分子机制研究

The acquired multidrug resistance during neoadjuvant chemotherapy severely inhibits the development of osteosarcoma treatment. It is important to fully clarify the mechanisms of drug resistance. Exosomes can play an important role in tumor microenvironment, and transmit chemoresistant information among tumor cells. It has been verified that in some tumors, exosome-mediated long noncoding RNAs（lncRNAs）are close related to the chemoresistance. Our preliminary data demonstrated that lncRNA antidifferentiation noncoding RNA（ANCR）was positively related to the cell proliferation of human osteosarcoma cells, and the high expression of lncRNA ANCR was detected both in the chemoresistant human osteosarcoma cells and in their exosomes. However, the molecular mechanism of exosomal lncRNA ANCR in chemoresistance of osteosarcoma has yet to be defined. In this study, we will reveal the important role of exosome in drug resistance of osteosarcoma, and the molecular mechanism of lncRNA ANCR in osteosarcoma-derived exosomes contributing to drug resistance, by gene silencing, RNA immunoprecipitation (RIP), RNA pull down, luciferase reporter gene assay and so on. Additionally，we will analyze the feasibility of using lncRNA ANCR in osteosarcoma-derived exosomes as a detective biomarker and therapeutic target for chemoresistance of osteosarcoma. These data will enrich theoretical foundations of anti-cancer drug resistance and broaden our horizons of anti-chemoresistance therapy in osteosarcoma.

新辅助化疗继发多药耐药已成为阻碍骨肉瘤疗效提升的关键。阐明化疗耐药机制具有重要意义。肿瘤细胞分泌的外泌体可在肿瘤细胞间传递耐药信息，在肿瘤微环境中扮演重要角色。部分肿瘤中，外泌体lncRNA已被证实与化疗耐药相关。我们前期研究发现lncRNA ANCR与人骨肉瘤细胞增殖密切相关，在耐药人骨肉瘤细胞及其外泌体中lncRNA ANCR呈现高表达。但外泌体lncRNA ANCR在骨肉瘤耐药中的作用及分子机制尚不明确。本课题将深入探究外泌体在骨肉瘤耐药中的作用，以外泌体lncRNA ANCR为研究对象，整合基因沉默、RIP、RNA pull down和荧光素酶报告基因等研究手段，以其调节的RNA结合蛋白、mRNA及miRNA为研究重心，探讨外泌体lncRNA ANCR在骨肉瘤耐药中的作用及分子机制，分析其作为耐药检测靶标及干预靶点的可行性。研究结果将丰富现有的耐药理论，拓展抗骨肉瘤耐药的治疗思维。

新辅助化疗继发多药耐药已成为阻碍骨肉瘤疗效提升的关键。阐明化疗耐药机制具有重要意义。肿瘤细胞分泌的外泌体可在肿瘤细胞间传递耐药信息，在肿瘤微环境中扮演重要角色。部分肿瘤中，外泌体lncRNA已被证实与化疗耐药相关。本课题已完成标书全部研究目标，明确外泌体在体外及体内参与骨肉瘤阿霉素耐药性调控，明确外泌体lncRNA ANCR调控体内及体外骨肉瘤阿霉素耐药性，发现人骨肉瘤细胞外泌体lncRNA ANCR与患者生存及阿霉素耐药性具有相关性，明确lncRNA ANCR可以通过ceRNA机制结合miR-33b-5p抑制靶基因CALML4表达水平。通过本课题资助，已发表SCI论文10篇，在投SCI论文1篇，培养硕、博士研究生2人。