SPINK4调控结肠癌细胞铁死亡的作用机制研究

Ferroptosis is a new form of cell death found in recent years, and also occur in the colon cancer cells, but the mechanism of colon cancer cells ferroptosis remains unclear. Evidence has confirmed that the change of cytokine is important to the development of colon cancer cells. Our previous study found that SPINK4 was specifically expressed in intestinal tissues, which was closely related to the clinicopathological stage and prognosis of colon cancer. SPINK4 also participates in the process of ferroptosis induced by Erastin in colon cancer cells and has a binding relationship with VIL1. However, the role of SPINK4 in the ferroptosis of colon cancer cells has not been reported. Combined with the results of previous studies, we propose the hypothesis that SPINK4 regulates the process of ferroptosis in colon cancer cells by VIL1. In order to verify this hypothesis, we will study the tissue, cellular, and animal model levels using siRNA silencing, co-immunoprecipitation and other techniques to study: (1) the role of SPINK4 in colon cancer cell phenotype; (2) SPINK4 induced Erastin (3) To clarify the interaction between SPINK4 and VIL1 in colon cancer cells, and to reveal the molecular mechanism of SPINK4 regulating VIL1 inhibition of ferroptosis in colon cancer cells. This project will uncover the mechanism which SPINK4 regulates ferroptosis in colon cancer cells, and provide new targets for the treatment.

铁死亡是一种新发现的细胞死亡方式，结肠癌细胞也存在铁死亡，但其机制尚未完全清楚。研究表明细胞因子在结肠癌发病过程中起重要作用，我们前期研究发现SPINK4特异性表达于肠道组织，与结肠癌的临床病理分期和预后密切相关；SPINK4参与了Erastin诱导结肠癌细胞铁死亡过程，且与VIL1存在结合关系，但迄今尚未有SPINK4在结肠癌细胞铁死亡中作用的报道。结合前期研究结果，我们提出假说，即SPINK4调控VIL1影响结肠癌细胞铁死亡。为了验证该假说，我们将从组织、细胞、动物水平进行研究，采用siRNA沉默、免疫共沉积等技术，阐明：（1）SPINK4对结肠癌细胞表型的作用；（2）SPINK4在Erastin诱导结肠癌细胞铁死亡中的作用；（3）结肠癌细胞中SPINK4与VIL1相互作用，进而影响结肠癌细胞铁死亡的分子机制。本项目将揭示SPINK4调控结肠癌细胞铁死亡新的机制，并为治疗提供新靶点。

本项目开展的SPINK4调控结肠癌细胞铁死亡的作用机制研究是以项目组前期通过临床大样本数据分析研究中发现的结直肠癌关键候选分子之一 SPINK4为切入点，结合目前细胞铁死亡在肿瘤发病和治疗的价值这一研究热点，探讨结直肠癌细胞铁死亡过程中SPINK4在其中的作用及其调控铁死亡的分子机制，为治疗晚期结肠癌提出新的治疗思路和筛选出有效的靶标分子。本项目按照研究计划开展了相关研究，本项目通过血清学实验、细胞实验、动物实验和细胞分子实验，检测结直肠癌患者和正常人组织中SPINK4和VIL1的表达水平。同时，构建小鼠结直肠癌模型。检测小鼠结直肠癌经Erastin治疗后SPINK4和VIL1的表达水平及铁死亡相关细胞因子GSH，GPX4的表达水平和铁含量；同时检测NF-κB和LCN2的表达水平，完成了预期的研究内容。本项目结果显示：1. SPINK4在结直肠癌组织中下降，促进大肠癌细胞增殖和转移；SPINK4经MYC-TP53途径抑制结直肠癌细胞铁死亡。2. VIL1促进结直肠癌的发生，VIL1经调控NF-κB /LCN2抑制抑制结直肠癌细胞铁死亡；3. 血清SPINK4水平对结直肠癌患者有较高的诊断价值，但无明显预后价值。项目完成时，共发表论文15篇，其中SCI论文13篇，获得广西医药卫生适宜技术推广奖二等奖1项，实用新型专利1项。培养硕士研究生2名。