LncRNA-KMU15在膀胱癌发生发展的作用及分子机制研究

Bladder cancer is the most common malignant tumor of urinary system, threatening to human health and social economic development. The main problems for the treatment of the cancer include， that early diagnosis are lack of specificity and sensitivity， and that a high rate of recurrence and malignant transformation occurs after the transitional operation treatment. It’s ignored that long noncoding RNA abnormal expression was closely correlated with tumor before a variety of lncRNAs were found associated with tumors, especially for some cancers the lncRNAs were unique. However, no lncRNAs are found playing roles in the occurrence and development of bladder cancer, let alone the specific lncRNAs for the tumor. In this study, lncRNA-KMU15 was first found and further demonstrated to be positively related to bladder cancer by lncRNA chip and RT-PCR etc. Although without any report on lncRNA-KMU15 internationally and domestically, our preliminary data prove lncRNA-KMU15 does be in close contact with bladder cancer. The further study will focus on exploring the mechanism of lncRNA-KMU15 in the bladder cancer pathogeneses, providing a new target and direction for the tumor intervention; and focus on exploring the use of lncRNA-KMU15 as indexes for early bladder cancer screening and prognosis monitoring.

膀胱癌是最常见的泌尿系统恶性肿瘤，其早期诊断技术缺乏特异性及灵敏性，而膀胱肿瘤部分切除术等治疗后复发率高、恶性转化程度强等是目前该领域存在的主要问题。研究发现，长链非编码RNA异常表达与肿瘤密切相关，现已发现与癌症相关的多种lncRNAs，并且许多癌症都有自己独特的lncRNA。然而，lncRNA对膀胱癌发生发展的影响知之甚少，是否存在膀胱癌特异的lncRNA也未见报道。本项目通过lncRNA表达芯片首次发现并进一步验证lncRNA-KMU15的表达量与膀胱癌呈正相关性，尽管目前国际国内尚没有任何关于lncRNA-KMU15的报道，我们初步的研究结果提示lncRNA-KMU15确实与膀胱癌有密切相关性。本研究将集中于探索lncRNA-KMU15参与膀胱癌的发病机制，为膀胱癌的干预治疗提供新的靶点和方向，探究以lncRNA-KMU15为指标实现膀胱癌早期筛查与预后监测的可能性。

背景：膀胱癌是最常见的泌尿系恶性肿瘤，其早期诊断技术缺乏特异性及灵敏性，我们前期研究发现lncRNA-KMU15与膀胱癌密切相关，因此将进一步探索lncRNA-KMU15参与膀胱癌的发病机制，为膀胱癌的治疗提供新的理论基础。.方向：主要研究在体外人类细胞模型中确定lncRNA-KMU15和癌症发生的关系，鉴定lncRNA-KMU15的蛋白作用网络及关键调节因子，探明lncRNA-KMU15调控该关键因子介导膀胱癌的分子机制，利用膀胱癌小鼠原位模型，研究lncRNA-KMU15治疗膀胱癌的可能性。.主要内容：通过LncRNA表达芯片检测发现与膀胱癌发生发展密切相关的lncRNA，lncRNA-KMU15对膀胱癌的发生、发展、转移及复发的影响及机制，并利用CO-IP和western blot深入探索lncRNA-KMU15调控膀胱癌进展的分子机制，构建lncRNA-KMU15稳定低表达的EJ细胞株，通过CCK-8、划痕实验、transwell侵袭实验以及流式细胞技术检测lncRNA-KMU15低表达对膀胱癌EJ细胞株生物学特性的影响；通过RT-qPCR检测发现lncRNA-KMU15的表达与膀胱癌的分级、分期的相关性；探讨lncRNA-KMU15通过与某miRNA竞争性结合调控某靶点对膀胱癌细胞化疗敏感性产生影响。.重要结果和关键数据：lncRNA-KMU15表达量与膀胱癌的发生呈正相关。lncRNA-KMU15敲低后，膀胱癌细胞的粘附、迁移及克隆形成等生物学功能均受到明显抑制，lncRNA-KMU15敲低后的荷瘤小鼠对DDP的化疗敏感性显著增强，其在DDP治疗下的生存率得到显著提升。RT-qPCR检测发现lncRNA-KMU15的表达与膀胱癌的分级、分期均呈显著正相关，并且lncRNA-KMU15高表达者总生存率明显低于lncRNA-KMU15低表达者。lncRNA-KMU15在膀胱癌ADM耐药株细胞Biu87-R中表达升高，并阐明了新型lncRNA-KMU15能够通过竞争性抑制miR-505-5P而促进c-Myc表达，进一步降低膀胱癌耐药株细胞对于ADM的化疗敏感性。.科学意义：通过上述实验发现，lncRNA-KMU15除和膀胱癌的发生发展相关外，还参与调控膀胱癌细胞的化疗耐药性的产生，这一发现为膀胱癌的治疗提供新的理论基础，为预防及治疗膀胱癌提供了全新的视角。