TBX5-AS1通过调节糖代谢抑制肺腺癌恶性进展的机制研究

Glucose metabolism reprogramming is a new hallmark of tumor cells. Through early tissue sequencing screening and bioinformatics analysis, we found that long non-coding RNA TBX5-AS1 is a tumor suppressor gene related to glucose metabolism in lung adenocarcinoma. Clinical samples verification demonstrated that TBX5-AS1 was significantly downregulated in lung adenocarcinoma tissues and correlated with the TNM staging and prognosis of patients. In vitro experiments showed that overexpression of TBX5-AS1 could inhibit the malignant phenotype and glucose metabolism reprogramming of lung adenocarcinoma. RIP experiment preliminarily proved that TBX5-AS1 could bind to the transcription factor RUNX2, while the RNA-seq results showed that SIRT6 varied most after the over-expression of TBX5-AS1. Meanwhile, the website predicted that RUNX2 could bind to the promoter region of SIRT6. Based on these results, we proposed a scientific hypothesis: in lung adenocarcinoma, TBX5-AS1 binds to RUNX2 and promotes the transcription of its downstream target gene SIRT6, regulates the tumor glucose metabolism, and thereby inhibits the malignant progression of lung adenocarcinoma. In this project, the mechanism of TBX5-AS1 inhibiting the malignant progression of lung adenocarcinoma through the regulation of glucose metabolism was studied from three aspects including in vivo, in vitro and clinical samples. The successful implementation of this project is expected to provide a new direction for the diagnosis and treatment of lung adenocarcinoma.

糖代谢重塑是肿瘤细胞的一种新标志。通过前期的组织测序筛选以及生物信息学分析，我们发现长链非编码RNA TBX5-AS1是肺腺癌中糖代谢相关的抑癌基因。临床标本验证显示TBX5-AS1在肺腺癌组织中显著低表达并且与病人的分期及预后相关；体外实验表明过表达TBX5-AS1可以抑制肺腺癌的恶性表型及糖代谢重塑。RIP实验初步证明TBX5-AS1可以与转录因子RUNX2结合，而过表达TBX5-AS1之后的测序结果显示去乙酰化酶SIRT6的上调最明显，网站预测提示RUNX2可以与SIRT6的启动子区结合。基于此我们提出科研假说：肺腺癌中，TBX5-AS1通过结合RUNX2，促进其对下游靶基因SIRT6的转录，从而抑制肺腺癌的糖代谢重塑以及恶性进展。本项目将从体内、体外、临床样本三个层面深入研究TBX5-AS1通过调节糖代谢抑制肺腺癌恶性进展的机制。本项目的成功实施有望为肺腺癌的诊治提供新的方向。

肺腺癌是肺癌最常见的类型，也是癌症相关死亡最常见的原因，预后较差。糖代谢重塑是肿瘤细胞的一种新标志，通过关键糖代谢相关酶的表达调节，使得肿瘤细胞更适应相对饥饿的环境。通过前期的组织测序筛选以及生物信息学分析，我们发现长链非编码RNA TBX5-AS1是肺腺癌中糖代谢相关的抑癌基因。临床标本验证显示TBX5-AS1在肺腺癌组织中显著低表达并且与病人的分期及预后相关；体内外实验表明过表达TBX5-AS1可以抑制肺腺癌的恶性表型及糖代谢重塑。RIP实及pulldown实验证明TBX5-AS1可以与转录因子RUNX2结合，而过表达TBX5-AS1之后的测序结果显示去乙酰化酶SIRT6的上调最明显，通过染色质免疫共沉淀实验证实，RUNX2可以与SIRT6的启动子区结合调节其转录，而TBX5-AS1能促进RUNX2对SIRT6的转录，并且使得受SIRT6抑制的关键糖酵解基因（GLUT1、PKM、HK1）下调。本项目的研究总结：肺腺癌中，TBX5-AS1通过结合转录因子RUNX2，促进其对下游靶基因SIRT6的转录，进而下调关键糖酵解基因（GLUT1、PKM、HK1）的表达，从而抑制肺腺癌的糖代谢重塑以及恶性进展。本项目从糖代谢重塑入手研究肿瘤细胞的恶性进展，具有较大的突破性，也进一步加深了人们对肺腺癌发生发展机制的认识。TBX5-AS1作为一个研究较少的lncRNA，有望作为临床诊断及预后的标记物，而RUNX2/SIRT6以及代谢酶（GLUT1、PKM、HK1）信号轴的研究，为临床的治疗提供了潜在靶点。