GOLPH3通过外泌体运输lncRNA-ATB促进膀胱癌化疗耐药的分子机制研究

Chemotherapy resistance is a major challenge for the treatment of bladder cancer. However, the potential mechanisms involved in chemotherapy resistance still remain unclear. Our previous study has demonstrated that exosome-mediated intercellular interaction was involved in the regulation of chemotherapy resistance. However, the generation of exosomes and molecular mechanism how exosomes regulate chemotherapy resistance are still unknown. Thereafter, we further found that high expression of GOLPH3 which is essential for exosomes generation was detected in chemotherapy resistant bladder cancer tissues. Significantly elevated lcnRNA-ATB was also found in exosomes secreted by chemotherapy resistant cancers. Therefore, we propose that GOLPH3 could promote the secretion of lncRNA-ATB encapsulated exosomes to surrounding cells to induce the chemotherapy resistance by enhancing the stemness of cancer cells. At first, we will validate the function of GOLPH3 in cancer tissues and cells. Furthermore, we plan to explore how GOLPH3 mediates the secretion of lncRNA-ATB encapsulated exosomes from resistant cells and transferring to the sensitive cells. Moreover, the mechanism how lncRNA-ATB mediates cancer cell stemness related genes will be illustrated. At last, we will analyze the clinical significance of the key molecules in chemotherapy resistant bladder cancer tissues. Taken together, this project will explore predictors and potential targets for the treatment of chemotherapy resistant bladder cancers.

膀胱癌化疗耐药是亟待解决的临床难题，但其具体机制不明。前期研究中我们发现外泌体介导的细胞间交流导致膀胱癌产生耐药，但外泌体的产生及其调控耐药机制并不清楚。后续发现调控外泌体的GOLPH3在耐药组织中高表达，且耐药癌细胞高表达的lncRNA-ATB在其外泌体内高度富集。所以本项目拟提出GOLPH3促进外泌体包装lncRNA-ATB并分泌进入相邻癌细胞，调控干性相关基因表达诱导耐药，从而扩大耐药细胞群体产生化疗耐药的理论设想。首先，我们拟在化疗病人样本和体内外耐药细胞模型中验证GOLPH3促进肿瘤细胞干性和化疗耐药；然后，研究GOLPH3如何诱导耐药膀胱癌分泌外泌体并运输lncRNA-ATB到相邻非耐药细胞；接着，阐述lncRNA-ATB调控肿瘤干性相关基因的分子机制；最后，分析关键分子的临床意义，并研究耐药膀胱癌的治疗。本项目有望筛选到化疗敏感性检测指标，并探索逆转膀胱癌化疗耐药的治疗策略。

化疗耐药的膀胱癌病人缺乏有效的临床治疗方法。本项目研究发现，miR34a/GOLPH3轴通过减少肿瘤干性来消除尿路上皮性膀胱癌的化疗耐药性。全转录组测序和功能研究，发现circSLC8A1作为miR-130b/miR-494的分子海绵，通过调控PTEN抑制膀胱癌进展。miR-30e-5p能通过靶向异粘蛋白MTDH抑制膀胱癌细胞的生长和侵袭。miR-582-FOXG1轴可能在细胞侵袭中起关键作用。应用蛋白质组学鉴定和分析膀胱癌组织和癌细胞的蛋白。我们鉴定到的很多癌细胞分泌蛋白已知和肿瘤转移和进展相关，并且筛选出的候选分泌蛋白可能和膀胱癌的进展相关。临床标本检测也显示，关键分子的高表达与临床预后不良相关。这些调控信号中的关键靶点可能作为膀胱癌化疗敏感性预测的标志物，并为临床耐药进展的膀胱癌治疗提供新的策略。