lncRNA-UCA1通过PKM2参与膀胱癌细胞Warburg效应的机制

Warburg effect is a hallmark of energy metalization in tumor cells. PKM2, highly expressed in many tumor tissues,is involved in glycolysis progress and promotes tumor Warburg effect. LncRNAs play a crucial role in tumor progress. In our previous studies, we screened the lncRNA-UCA1 interaction proteins, including Keratin 10 and PKM2. The interaction between UCA1 and K10 could active PI3K/AKT/mTOR signaling pathway. HIF-1α could up-regulate UCA1 expression in bladder cancer cells. Intriguingly, ectopic expression of UCA1 increased glycose consumption and lactatic acid production, suggesting that UCA1 could participate in tumor Warburg effect. Accordingly, in this study we will further confirm the interaction between UCA1 and PKM2 with RNA binding protein immunoprecipitation, charaterize the binding domain of PKM2, and explore the bio-function and mechanism of Warburg effect mediated by the interaction between UCA1 and PKM2. Moreover, we will research PKM2 expression regulated by the PI3K/AKT/mTOR signailing actived with UCA1 and K10 interation. Through this study, we will first illuminate the bio-funtion and mechanism of lncRNAs involved in Warburg efffect. These studies will be helpful to find the new specific tumor therapeutic target from tumor cell metalization and proliferation.

Warburg效应是肿瘤细胞能量代谢的重要特征；PKM2参与糖酵解过程，在许多肿瘤组织中表达增高，促进肿瘤细胞Warburg效应；lncRNAs在肿瘤发展中起着重要作用。我们前期研究发现lncRNA-UCA1能与Keratin10和PKM2结合；UCA1与K10结合后激活PI3K/AKT/mTOR信号通路；HIF-1α可促进膀胱癌细胞UCA1转录，异位表达UCA1使葡萄糖消耗和乳酸生成增加，表明UCA1参与调节肿瘤细胞Warburg效应。据此本项目拟采用RNA结合蛋白免疫沉淀法验证UCA1与PKM2相互作用，通过体内外实验和组织标本检测，探讨两者对Warburg效应的作用及原理；并深入研究UCA1与K10结合激活PI3K/AKT/mTOR信号通路对PKM2表达的影响。从而率先阐明lncRNAs对肿瘤细胞Warburg效应的作用及其分子机制，为寻找肿瘤细胞代谢和增殖特异性治疗新靶点奠定基础。

lncRNAs在肿瘤发展中起着重要作用。Warburg效应是肿瘤细胞能量代谢的重要特征。我们前期研究发现lncRNA-UCA1参与调节膀胱癌细胞Warburg效应。据此，本项目通过体内外实验发现UCA1通过mTOR/STAT3/HK2通路以及mTOR/miR143/HK2通路促进膀胱癌细胞Warburg效应，发现UCA1通过miR-145-ZEB1/2-FSCN1通路促进膀胱癌细胞上皮-间质转化从而推动癌细胞侵袭迁移；利用CRISPR/Cas9技术敲除UCA1，证实UCA1促进膀胱癌恶性增殖、迁移和侵袭；等重同位素多标签相对定量蛋白质组学（iTRAQ）技术检测UCA1敲除对蛋白表达水平的影响，共发现了139个差异表达的蛋白分子，其中上调1.5倍的有66个蛋白分子，下调1.5倍的有73个分子；利用CRISPR/Cas9技术联合敲除UCA1和PD1通过调节肿瘤免疫微环境、对膀胱癌具有潜在治疗价值。本课题研究了lncRNA UCA1对膀胱癌Warburg效应的作用及其分子机制，进一步探寻了相关的节点分子，并结合最新的基因编辑技术与免疫检查点调控策略、尝试建立靶向UCA1的膀胱癌治疗新策略，为膀胱癌进展的分子机制研究与临床治疗策略奠定了良好基础。