恶性肿瘤中糖酵解增强，为其无限制增殖提供能量、生物大分子前体及抗氧化物质。丙酮酸激酶(PKM2)是肿瘤糖酵解途径中的限速酶之一，受多种翻译后修饰作用通过改变其活性调控代谢重编程。一氧化氮合酶(NOS)催化一氧化氮(NO)合成，通过亚硝化(S-nitrosylate)修饰靶蛋白促进肿瘤恶性进展。本课题组前期实验证明nNOS在卵巢癌中高表达，可结合并亚硝化PKM2。因此，我们推测nNOS通过亚硝化PKM2促进肿瘤糖酵解和细胞增殖。为此，本研究旨在1）利用质谱技术鉴定出PKM2亚硝化位点；2）构建PKM2亚硝化突变的卵巢癌细胞株，3）体内、外水平研究PKM2亚硝化对糖代谢流和细胞增殖的调控作用及机制；4）临床分析PKM2亚硝化与卵巢癌患者进展预后的关系并筛选亚硝化差异的代谢酶谱。本课题将探讨nNOS诱导的PKM2亚硝化对卵巢癌糖酵解和细胞增殖的调控作用和机制，为寻找卵巢癌诊断及治疗提供理论依据。

One of the fundamental properties of cancer cells is that they underwent metabolic reprogramming from oxidative phosphorylation to aerobic glycolysis during carcinogenesis. Pyruvate kinase M2 (PKM2), a rate-limiting enzyme in the last step of glycolysis, exerts an important role in glycolysis and tumor malignancy. Our previous work showed that neural nitric oxide synthases (nNOS) is upregulated in human ovarian cancer and induces PKM2 S-nitrosylation, a reaction that coupling of a mitroso moiety to a reactive thiol group in specific cysteine residues leading to the formation of an S-nitrosothiol (SNO). However, the functional and clinical significance of PKM2 S-nitrosylation has not been studied. Here we aim to explore the role of nNOS induced S-nitrosylation of PKM2 on glycolysis and ovarian cancer progression. The specific aims are: 1) identify the S-nitrosylating site of PKM2 through mass spectrum method. 2) establish PKM2-knockout ovary cancer cell lines overexpressing S-nitrosylation mimetic- or deficient PKM2 cDNA plasmid by using CRISPR-Cas9 and site-directed mutagenesis technologies. 3) analyze the role of PKM2 S-nitrosylation on glycolysis flow in SNO-mutant ovary cancer cells with GS-MS methods. 4) study the role of PKM2 S-nitrosylation on PKM2 function in vitro and in vivo. 5) explore prognostic and diagnostic roles of PKM2 S-nitrosylation in ovarian cancer patients. The proposed research will not only lead to a better understanding of NOS-induced PKM2 S-nitrosylation on tumor progression, but also provide potential prognosticator for ovarian cancer.