PKLR基因调控FOXO4影响DNA复制促进肝癌细胞增殖的作用及机制研究

Pyruvate kinase has close relationship with hepatocellular carcinoma (HCC). Recently, researchers found the pyruvate kinase might act as a signal molecule to affect the biological behaviors of HCC with unclear mechanism. We firstly proposed that the pyruvate kinase L/R (PKLR) gene is liver specifically expressed potential target for HCC treatment for its activity on promoting cell proliferation. PKLR gene negatively regulated the FOXO4 transcription factor, significantly upregulated the expression of key genes (CDC6, PCNA and PAF15) in DNA replication; FOXO4 had potential binding sites with key genes on DNA replication. Therefore, we hypothesized that PKLR might promote the HCC proliferation by reducing the resistance of DNA replication via down-regulation of FOXO4 expression. The purpose of this study is to investigate the mechanism of PKLR gene on promoting the proliferation of HCC cells via regulation of FOXO4 transcription factor and upregulation of genes in DNA replication. The clinical value of PKLR will also be assessed in HCC patients. We will use plasmid transfection, confocal imaging and co-immunoprecipitation techniques etc to test the samples from hepatic cell line, xenografts of nude mice and clinical pathological tissues. And this project will provide new concepts for targeted HCC treatment.

丙酮酸激酶与肝癌的发生发展关系密切。近年来有研究发现丙酮酸激酶可作为信号分子影响肝癌生物学行为，但具体机制不详。我们的前期研究发现，丙酮酸激酶基因L/R(PKLR)在肝脏特异表达，促进细胞增殖，是肝癌治疗的潜在靶点。PKLR基因负向调控FOXO4转录因子,显著提高肝癌细胞DNA复制关键基因（CDC6、PCNA和PAF15）的表达水平；且FOXO4存在与DNA复制关键基因的潜在结合位点。据此我们提出假说：PKLR通过下调FOXO4的表达，减轻DNA复制阻力，促进肝癌细胞增殖。本项目拟利用肝癌细胞株、裸鼠原位移植瘤模型及临床组织标本，以质粒转染、共聚焦成像及免疫共沉淀等技术，研究PKLR基因调控FOXO4转录因子促进下游DNA复制和肝癌细胞增殖的作用及分子机制，并在肝癌人群中验证其临床价值，为肝癌靶向治疗提供新的思路。

丙酮酸激酶与肝细胞增殖和再生的发生发展关系密切。 近年来有研究发现丙酮酸激酶L/R (PKLR)可作为信号分子影响肝细胞及肝癌细胞的生物学行为，但具体机制不详。我们发现：供肝脂肪变性在中国肝移植病例中非常普遍，PKLR与供肝脂肪变性相关; PKLR降低了移植术后不良预后发生的风险; PKLR的下调也将带动引起更多的并发症，导致更差的预后; PKLR表达水平对移植后并发症的影响独立于受者术后的峰值胆红素和转氨酶水平。我们通过对脂肪变供肝的代谢组学分析后发现：甘油磷脂通路上的磷脂酰胆碱（PC）和磷脂酰乙醇胺（PE）同时影响了供肝大泡型脂肪变性和移植肝衰竭的发生。PC和PE的下调在显著增加大泡性脂肪肝发生的同时也促进了移植肝衰竭的发生。通过对转染肝细胞的RNA测序和靶向代谢组学分析，构建PKLR相关的转录-代谢调控网络后发现：PKLR的表达也与供肝磷脂酰胆碱的表达水平显著相关。本项目通过临床组织样本及体内/体外模型，从临床流行病学，系统生物学，分子生物学水平证实PKLR基因在肝细胞再生，脂质从头合成及肝癌细胞增殖等方面的重要意义，为后续转化研究提供了重要的理论依据。

内容获取失败，请点击重试