声带白斑组织SMAD4、IL-35的表达及其与喉癌的关系研究

Expression of SMAD4 and IL-35 in vocal cord leukoplakia and their relationship with prognosis

目的分析声带白斑组织果蝇母亲DPP同源物4(SMAD4)、IL-35表达情况,并探讨其与喉癌的关系。方法选取2019年1月至2021年1月嘉兴市第一医院耳鼻咽喉科收治的行手术治疗的声带白斑患者30例为声带白斑组,声带息肉患者30例为声带息肉组,喉癌患者30例为喉癌组,采用RT-PCR法、免疫荧光染色法检测声带息肉组织、白斑组织及喉癌组织SMAD4、IL-35(包括2个亚型:IL-12p35、Ebi3)mRNA和蛋白的表达情况;分析声带组织SMAD4 mRNA表达水平、蛋白阳性表达率与上皮异常增生程度的相关性;分析声带白斑患者术后发展为喉癌的危险因素;分析影响喉癌患者预后的危险因素。结果声带息肉组织SMAD4 mRNA表达水平、蛋白阳性表达率>声带白斑组织>喉癌组织(均P<0.05),声带息肉组织IL-12p35与Ebi3 mRNA表达水平、蛋白阳性表达率<声带白斑组织<喉癌组织(均P<0.05)。声带组织SMAD4 mRNA表达水平、蛋白阳性表达率与声带组织上皮异常增生程度均呈负相关(r_(s)=-0.637、-0.817,均P<0.05);IL-12p35 mRNA表达水平、蛋白阳性表达率与声带组织上皮异常增生程度均呈正相关(r_(s)=0.712、0.673,均P<0.05);Ebi3 mRNA表达水平、蛋白阳性表达率与声带组织上皮异常增生程度均呈正相关(r_(s)=0.681、0.776,均P<0.05)。Cox多因素分析显示,声带白斑形态分型差、上皮异常增生程度高、SMAD4低表达及Ebi3高表达是声带白斑患者术后发展为喉癌的独立危险因素(均P<0.05);淋巴结转移及IL-35高表达是影响喉癌患者预后的独立危险因素(均P<0.05)。结论SMAD4与IL-35的表达变化可能是喉癌发生、发展的早期事件,SMAD4低表达及IL-35高表达可能通过影响信号通路或免疫抑制影响声带白斑的转归,两者或可作为评估声带白斑患者转归及喉癌患者预后的指标。

Objective To investigate the expression of SMAD4 and IL-35 in vocal cord leukoplakia and their relation with carcinogenesis.Methods From January 2019 to January 2021,30 patients with vocal cord leukoplakia,30 patients with vocal cord polyps and 30 patients with laryngeal cancer who underwent surgical treatment in the Department of Otorhinolaryngology,Jiaxing First Hospital were enrolled in the study.Tissue samples were collected and the mRNA and protein expressions of SMAD4 and IL 35(including IL 12p35 and Ebi3 subtypes)were detected by RT-PCR and immunofluo-rescence staining respectively.The correlation expression of SMAD4 and IL 35 subtypes with the degree of epithelial dysplasia was analyzed.The risk factors of laryngeal cancer in patients with vocal cord leukoplakia after operation were analyzed.Results The SMAD4 mRNA expression level and protein expression rate in vocal cord polyps were higher than those in vocal cord leukoplakia,while values in the later were higher than those in laryngeal carcinoma(all P<0.05).The IL 12p35 and ebi3 mRNA expression level and protein positive expression rate in vocal cord polyps were lower than those in vocal cord leukoplakia,while the values in the later were lower than those in laryngeal carcinoma(all P<0.05)..SMAD4 mRNA expression level and protein positive expression rate were negatively correlated with the degree of abnormal hyperplasia of vocal cord epithelium(r_(s)=-0.637,-0.817,both P<0.05);IL-12p35 mRNA expression level and protein positive expression rate were positively correlated with the degree of vocal cord epithelial dysplasia(r_(s)=0.712,0.673,both P<0.05);Ebi3 mRNA expression level and protein positive expression rate were positively correlated with the degree of vocal cord epithelial dysplasia(r_(s)=0.681,0.776,both P<0.05).Multivariate Cox analysis showed that poor histological classification of vocal cord leukoplakia,high degree of epithelial dysplasia,low expression of SMAD4 and high expression of Ebi3 were independent risk factors for laryngeal cancer in patients with vocal cord leukoplakia after operation(all P<0.05).Lymph node metastasis and high expression of IL 35 were independent risk factors affecting the prognosis of laryngeal cancer patients(both P<0.05).Conclusion SMAD4 and IL 35 may be associated with the occurrence and development of laryngeal carcinoma;low expression of SMAD4 and high expression of IL-35 may affect the prognosis of vocal cord leukoplakia through signal pathway or immunosuppression.SMAD4 and IL-35 may be used as target indicators to evaluate the prognosis of vocal cord leukoplakia and laryngeal carcinoma.