PTEN基因甲基化在喉癌中的临床价值及对喉癌细胞生物学行为的影响  

Clinical value of PTEN gene methylation in laryngeal carcinoma and its effect on the biological behavior of laryngeal carcinoma cells

作　　者：王爱华 凡丹丹 吴俊叶 石亮亮 李鹏程[2] WANG Aihua;FAN Dandan;WU Junye(Department of Otolaryngology,Wuhan Third Hospital,Hubei,Wuhan 430074,China)

机构地区：[1]湖北省武汉市第三医院耳鼻喉科,430074 [2]华中科技大学同济医学院附属协和医院肿瘤中心

出　　处：《河北医药》2025年第3期363-367,374,共6页Hebei Medical Journal

基　　金：湖北省自然科学基金(编号:2020HBA216)。

摘　　要：目的研究磷酸酯酶与张力蛋白同源物(PTEN)基因甲基化在喉癌中的临床价值及对喉癌细胞生物学行为的影响。方法选择2016年1月至2017年12月诊治的喉癌患者(LGC组)和喉良性疾病患者为研究对象。分别将寡核苷酸序列(MON、UMON和CON)并转染至Hep-2喉癌细胞。采用甲基特异性PCR检测PTEN基因甲基化状态。分析PTEN基因甲基化与临床病理因素的关系。ROC曲线分析PTEN基因甲基化预测喉癌死亡效能。单因素及多因素Logistics回归分析喉癌患者死亡的危险因素。Kaplan-Meier分析PTEN基因甲基化与生存期的关系。CCK-8检测喉癌细胞增殖。细胞划痕试验检测细胞迁移。Transwell小室检测细胞侵袭数。结果LGC组患者肿瘤组织PTEN基因甲基化率显著高于癌旁组织和对照组(分别为72.92%、8.33%、8.00%),差异有统计学意义(χ^(2)=108.215,P<0.01)。TU138、TU212喉癌细胞中PTEN基因为甲基化状态,而在Hep-2、NHLEC细胞中为未甲基化状态。低分化、肿瘤最大直径≥3 cm、有淋巴结转移及TNM分期Ⅲ+Ⅳ期患者PTEN基因甲基化率高于中-高分化、肿瘤最大直径<3 cm、无淋巴结转移及TNM分期Ⅰ+Ⅱ期患者(P<0.05)。PTEN基因甲基化预测喉癌患者死亡的敏感度、特异度、AUC均高于肿瘤分化程度、肿瘤最大直径、淋巴结转移、TNM分期(Z/P=3.267/0.016、4.022/0.007、3.728/0.011、3.452/0.013)。PTEN基因甲基化、肿瘤分化程度低分化、肿瘤最大直径≥3 cm、有淋巴结转移、TNM分期Ⅲ+Ⅳ期为喉癌死亡的独立危险因素(P<0.05)。转染MON后Hep-2喉癌细胞PTEN基因甲基化,转染UMON及CON后Hep-2喉癌细胞PTEN基因未甲基化。MON组Hep-2喉癌细胞增殖活力、细胞迁移数及侵袭数显著高于UMON组及CON组(P<0.05)。结论PTEN基因甲基化可促进喉癌细胞增殖、迁移及侵袭。PTEN基因高甲基化与喉癌病情及预后密切相关,可作为喉癌病情及预后评估的标志物。Objective To study the clinical value of phosphatase and tensin homolog(PTEN)gene methylation in laryngeal carcinoma and its effect on the biological behavior of laryngeal carcinoma cells.Methods Patients with laryngeal carcinoma(LGC group)and patients with benign laryngeal diseases diagnosed and treated from Jan 2016 to Dec 2017 were selected as the research subjects.The oligonucleotide sequences(MON,UMON and CON)were transfected into the laryngeal carcinoma cell line Hep-2.Methylation-specific PCR(MSP)was used to detect the methylation status of PTEN gene.The correlation between PTEN gene methylation and clinicopathological factors of laryngeal carcinoma was identified.Receiver operating characteristic(ROC)curves were depicted to analyze the potential of PTEN methylation in predicting the mortality of laryngeal carcinoma.Univariate and multivariate Logistic regression analyses were conducted to analyze risk factors for the mortality of laryngeal carcinoma.Kaplan-Meier survival analysis was performed to illustrate the relationship between PTEN gene methylation and survival.Cell proliferation,migration and invasion were examined by cell counting kit-8(CCK-8)assay,wound healing assay and Transwell assay,respectively.Results The methylation rate of PTEN gene in LGC tissues was significantly higher than that of the paracancerous tissues and controls(72.92%vs.8.33%vs.8.00%,χ^(2)=108.215,P<0.01).The PTEN gene was methylated in the laryngeal carcinoma cell lines TU138 and TU212,but unmethylated in Hep-2 and NHLEC cells.The PTEN gene methylation rate was significantly higher in LGC patients with poor differentiation,maximum tumor diameter≥3cm,lymph node metastasis,and TNM stagingⅢ+Ⅳthan those of counterparts(P<0.05).The sensitivity,specificity,and area under the curve(AUC)of PTEN gene methylation in predicting the mortality of laryngeal carcinoma were all significantly higher than those of the degree of tumor differentiation,maximum tumor diameter,lymph node metastasis,and tumor-node-metastasis(TNM)staging(Z/P=3.

关 键 词：喉癌 张力蛋白同源物 甲基化 临床价值 生物学行为 

分 类 号：R739.65[医药卫生—肿瘤]