【摘要】 背景：研究发现脂肪酸诱导的铁死亡抵抗是肿瘤淋巴结转移的重要机制之一。目的：探讨小核仁RNA宿主基因16（SNHG16）对食管癌细胞铁死亡的影响及其可能的作用机制。方法：纳入2024年9月—2025年3月东南大学附属中大医院收治的63例食管癌患者，采用RT-qPCR检测SNHG16 mRNA表达水平，分析其与食管癌患者临床病理特征间的关系。培养人食管癌细胞株Eca109、KYSE410，并构建SNHG16过表达和敲减细胞株，分别设置过表达SNHG16组和过表达阴性对照组、敲减SNHG16组和敲减阴性对照组。采用流式细胞术检测细胞内脂质活性氧（ROS）水平，比色法检测细胞内Fe²⁺水平，气相色谱-质谱联用技术检测非靶向脂肪酸水平，蛋白质印迹法检测硬脂酰辅酶A脱饱和酶1（SCD1）蛋白表达水平。结果：食管癌组织中SNHG16 mRNA表达水平显著高于癌旁正常组织（P<0.000 1），其表达水平与食管癌淋巴脉管浸润显著相关（P=0.002），与患者的性别、年龄、肿瘤分化程度、部位、长径、TNM分期无关（P均>0.05）。过表达SNHG16后，Eca109细胞内脂质ROS和Fe²⁺水平显著低于阴性对照组（P均<0.05）。敲减SNHG16表达后，KYSE410细胞内脂质ROS和Fe²⁺水平显著高于阴性对照组（P均<0.05）。敲减SNHG16表达后，KYSE410细胞内中、长链不饱和脂肪酸浓度较阴性对照组降低，主要为油酸（C18:1n9c）、棕榈酸（C16:0）、亚油酸（C18:2n6c）等（P均<0.05）。过表达SNHG16后，Eca109细胞中SCD1蛋白表达水平显著高于阴性对照组（P=0.002 8）；敲减SNHG16表达后，KYSE410细胞中SCD1蛋白表达水平显著低于阴性对照组（P=0.001 6）。结论：SNHG16可能通过上调SCD1表达调控脂肪酸代谢，抑制食管癌细胞铁死亡，从而在食管癌淋巴结转移中发挥重要作用。更多还原

【Abstract】 Background: Studies have revealed that fatty acid-induced ferroptosis resistance is a crucial mechanism underlying tumor lymph node metastasis. Aims: To investigate the effect of small nucleolar RNA host gene 16(SNHG16) on ferroptosis in esophageal cancer cells and its potential mechanism. Methods: A total of 63 esophageal cancer patients admitted to Zhongda Hospital, Southeast University from September 2024 to March 2025 were enrolled. The expression level of SNHG16 mRNA was detected by RT-qPCR, and its correlation with patients′ clinicopathological characteristics was analyzed. Human esophageal cancer cell lines Eca109 and KYSE410 were cultured in vitro to construct SNHG16 overexpression or knockdown cell lines. Cell lines transfected with negative control lentivirus for gene overexpression or for gene knockdown were also constructed. Intracellular lipid reactive oxygen species(ROS) level was measured by flow cytometry, intracellular Fe2+ level by colorimetry, untargeted fatty acid levels by gas chromatography-mass spectrometry, and protein expression level of stearoyl-CoA desaturase 1(SCD1) by Western blotting. Results: The expression level of SNHG16 mRNA in esophageal cancer tissues was significantly higher than that in adjacent normal tissues(P<0.000 1), and its expression level was significantly correlated with lymphovascular invasion(P=0.002), but not with gender, age, tumor differentiation, location, longest diameter, or TNM stage(all P>0.05). Overexpression of SNHG16 in Eca109 cells resulted in significantly lower levels of lipid ROS and Fe2+ compared to negative control group(all P<0.05). Conversely, knockdown of SNHG16 in KYSE410 cells led to significantly higher levels of lipid ROS and Fe2+ compared to negative control group(all P<0.05). Knockdown of SNHG16 reduced the concentration of medium-and long-chain unsaturated fatty acids in KYSE410 cells compared to negative control group, mainly including oleic acid(C18:1n9c), palmitic acid(C16:0), linoleic acid(C18:2n6c), etc.(all P<0.05). Overexpression of SNHG16 significantly upregulated SCD1 protein expression in Eca109 cells(P=0.002 8), whereas knockdown of SNHG16 significantly downregulated SCD1 protein expression in KYSE410 cells(P=0.001 6). Conclusions: SNHG16 may regulate fatty acids metabolism by upregulating SCD1 expression, thereby inhibiting ferroptosis in esophageal cancer cells. This mechanism plays an important role in lymph node metastasis of esophageal cancer.更多还原