Src heterodimerically activates Lyn or Fyn to serve as targets for the diagnosis and treatment of esophageal squamous cell carcinoma
Src heterodimerically activates Lyn or Fyn to serve as targets for the diagnosis and treatment of esophageal squamous cell carcinoma
复制标题
DOI：
10.1007/s11427-022-2216-x
复制
发表时间：
2023-02
期刊：
影响因子：
--
通讯作者：
Jing Zhang;Di Zhao;Lingyuan Zhang;Yuanfan Xiao;Qingnan Wu;Yan Wang;Jie Chen;Q. Zhan
中科院分区：
文献类型：
--
作者：
Jing Zhang;Di Zhao;Lingyuan Zhang;Yuanfan Xiao;Qingnan Wu;Yan Wang;Jie Chen;Q. Zhan
尽管Src是最古老和研究最多的癌蛋白之一，但其在恶性肿瘤中的功能仍有待进一步确定。在这项研究中，我们证明了ponatinib对Src活性的抑制可以有效抑制体外和体内几种食管鳞状细胞癌（ESCC）的恶性表型，而对正常食管上皮细胞（NEECs）没有生长抑制作用。重要的是，我们结合了磷蛋白组学和几种细胞和分子生物学策略，以确定Src与Src家族激酶（SFKs）成员（如Fyn或Lyn）相互作用，形成异源二聚体。Src与Fyn和Lyn的相互作用使SH2 （Fyn tyr185或Lyn Tyr183）和激酶结构域（Fyn tyr420或Lyn Tyr397）中的酪氨酸位点磷酸化，这对ESCC的发展至关重要。而Src在NEECs中不能与Fyn或Lyn形成异源二聚体。我们使用RNA测序全面证明Src活性的抑制有效阻断了几种关键的肿瘤促进途径，如JAK/STAT、mTOR、干细胞相关和代谢相关途径。实时聚合酶链反应（RT-PCR）结果证实，Lyn和Fyn是src介导的肿瘤生长或转移相关分子表达的关键效应因子。此外，临床ESCC样本结果显示pSrc Tyr419、Fyn tyr185或Tyr420、Lyn tyr183或tyr397的高激活可能是ESCC预后的生物标志物。本研究表明Src/Fyn和Src/Lyn异二聚体可作为ESCC治疗的靶点。
Although Src is one of the oldest and most investigated oncoproteins, its function in tumor malignancy remains to be defined further. In this study, we demonstrated that the inhibition of Src activity by ponatinib effectively suppressed several malignant phenotypes of esophageal squamous cell carcinoma (ESCC) bothin vitroandin vivo, whereas it did not produce growth-inhibitory effects on normal esophageal epithelial cells (NEECs). Importantly, we combined phosphoproteomics and several cellular and molecular biologic strategies to identify that Src interacted with the members of Src-family kinases (SFKs), such as Fyn or Lyn, to form heterodimers. Src interactions with Fyn and Lyn phosphorylated the tyrosine sites in SH2 (Fyn Tyr185or Lyn Tyr183) and kinase domains (Fyn Tyr420or Lyn Tyr397), which critically contributed to ESCC development. By contrast, Src could not form heterodimers with Fyn or Lyn in NEECs. We used RNA sequencing to comprehensively demonstrate that the inhibition of Src activity effectively blocked several critical tumor-promoting pathways, such as JAK/STAT, mTOR, stemness-related, and metabolism-related pathways. Results of the real-time polymerase chain reaction (RT-PCR) assay confirmed that Lyn and Fyn were critical effectors for the Src-mediated expression of tumor growth or metastasis-related molecules. Furthermore, results of the clinical ESCC samples showed that the hyperactivation of pSrc Tyr419, Fyn Tyr185or Tyr420, and Lyn Tyr183or Tyr397could be biomarkers of ESCC prognosis. This study illustrates that Src/Fyn and Src/Lyn heterodimers serve as targets for the treatment of ESCC.