具核梭杆菌对食管鳞状细胞癌铁死亡相关蛋白表达及预后的影响

Effect of Fusobacterium nucleatum regulating ferroptosis related protein expression on prognosis of esophageal squamous cell carcinoma

目的:探讨具核梭杆菌(Fn)对食管鳞状细胞癌(ESCC)铁死亡相关蛋白缺氧诱导因子-1α(HIF-1α)及谷胱甘肽过氧化物酶4(GPX4)表达及预后的影响。方法:将ESCC细胞KYSE30及KYSE150各分为4组:对照组(不感染Fn),Fn感染12、24、48 h组。采用试剂盒检测细胞中活性氧(ROS)与丙二醛(MDA)含量,CCK8法检测顺铂(CDDP)对KYSE30和KYSE150细胞的半数抑制浓度(IC 50),Western blot法检测HIF-1α与GPX4蛋白表达水平。采用RNAscope技术检测222例ESCC及其癌旁正常食管组织中Fn感染情况,采用免疫组化法检测HIF-1α与GPX4蛋白的表达情况。采用Kaplan-Meier法绘制Fn感染阳性和阴性ESCC患者术后生存曲线。结果:Fn感染可诱导KYSE30及KYSE150细胞HIF-1α和GPX4蛋白高表达,提高细胞的氧化应激水平,并降低细胞对CDDP的应答效力,具有时间依赖性(P<0.05)。ESCC组织中Fn感染,HIF-1α及GPX4蛋白表达的阳性率均高于癌旁正常食管组织(P<0.05)。ESCC患者中男性、有吸烟史、有饮酒史、肿瘤低分化、肿瘤浸润深度侵及纤维膜、有淋巴结转移、TNM分期Ⅲ/Ⅳ期者Fn感染阳性率增加(P<0.05)。Fn感染阳性的ESCC患者术后生存状况较Fn感染阴性者差(P<0.001)。结论:Fn感染可抑制ESCC铁死亡,可能与ESCC患者预后不良有关。

Aim:To investigate the effect of Fusobacterium nucleatum(Fn)on the expressions of ferroptosis related protein hypoxia-inducible factor-1α(HIF-1α)and glutathione peroxidase 4(GPX4)in esophageal squamous cell carcinoma(ESCC)and prognosis of ESCC patients.Methods:ESCC KYSE30 and KYSE150 cells were allocated into 4 groups:control group,Fn infection group for 12 hours group,Fn infection group for 24 hours group and Fn infection group for 48 hours group.ROS and MDA kits were used to detect the content of oxidative stress indexes in cells.IC 50 of cisplatin(CDDP)on KYSE30 and KYSE150 cells was detected by CCK8 kit,and the expression levels of HIF-1αand GPX4 were detected by Western blot.Fn infection in 222 cases of ESCC and adjacent normal esophageal tissues were detected by RNAscope technique,and the expression of HIF-1αand GPX4 was detected by immunohistochemistry.Kaplan-Meier method was used to draw survival curves of ESCC patients with positive or negative Fn infection.Results:Fn infection induced the high expressions of HIF-1αand GPX4 protein in KYSE30 and KYSE150 cells,enhanced the level of oxidative stress,and decreased the response to CDDP in a time-dependent manner(P<0.05).The positive rates of Fn infection,HIF-1αand GPX4 protein expression in ESCC tissue were higher than those in adjacent normal esophageal tissue(P<0.05).In ESCC patients,the positive rate of Fn infection was increased in males,and patients with smoking history,drinking history,low tumor differentiation,tumor invasion depth in fibrous membrane,lymph node metastasis,TNM stageⅢ/Ⅳ(P<0.05).The survival of ESCC patients with Fn infection was worse than that without Fn infection after operation(P<0.001).Conclusions:Fn infection can induce ferroptosis of ESCC,which may be associated with poor prognosis of ESCC patients.