宫颈癌组织FBN1 基因甲基化状态与临床病理特征及预后的相关性研究

目的 探究宫颈癌（cervical cancer,CC）组织原纤维蛋白-1（fibrillin-1,FBN1）基因甲基化状态与患者临床病理特征及预后的关系。方法 选取2014 年6 月~2017 年4 月邯郸市第一医院诊治的98 例CC 患者为研究对象,收集经手术切除的CC 组织、癌旁组织。甲基化特异性PCR 法测定CC 组织、癌旁组织FBN1 基因甲基化状态;蛋白印迹法检测CC 组织、癌旁组织FBN1 蛋白表达水平;对CC 患者进行为期5 年的随访,记录患者生存情况;比较CC 组织和癌旁组织FBN1 基因甲基化发生率以及FBN1 基因非甲基化组和FBN1 基因甲基化组CC 组织FBN1 蛋白表达水平;分析CC 组织FBN1 基因甲基化状态与患者临床病理特征的关系、FBN1 基因甲基化状态与患者预后的关系以及CC 患者预后的影响因素。结果 CC 组织FBN1 基因甲基化发生率（60.20%）高于癌旁组织（12.24%）,差异有统计学意义（χ2=48.785,P<0.05）;FBN1 基因甲基化组FBN1 蛋白表达水平（0.61±0.12）低于FBN1 基因非甲基化组（1.59±0.32）,差异有统计学意义（t=21.401,P<0.05）。CC组织FBN1基因甲基化状态与患者TNM分期、高危型人乳头瘤病毒DNA（highrisk-human papillomavirus DNA,HR-HPV DNA）、淋巴结转移相关（χ2=7.578,8.140,7.814,均P<0.05）;FBN1 基因甲基化组CC 患者5 年累积生存率为38.98%,低于FBN1 基因非甲基化组（76.92%）,差异有统计学意义（χ2=13.464,P<0.05）。HR-HPV DNA 阳性[OR（95%CI）: 2.534（1.577~4.072）],有淋巴结转移[OR（95%CI）: 2.426（1.546~3.808）],TNM 分期Ⅲ ~ Ⅳ期[OR（95%CI）: 2.702（1.633~4.471）] 和FBN1 基因甲基化[OR（95%CI）: 2.394（1.531~3.743）]是影响CC 患者预后的独立危险因素（P<0.05）。结论 CC 患者癌组织中FBN1 基因甲基化水平较高,其与TNM 分期、HR-HPV DNA 和淋巴结转移等临床病理特征及预后相关,为临床诊治CC 和评估CC 患者预后提供新方向。

Correlation between Methylation Status of FBN1 Gene in Cervical Cancer Tissue and Clinicopathological Features and Prognosis

Objective To explore the relationship between the methylation status of fibrillin-1 (FBN1) gene and the clinicopathological characteristics and prognosis of cervical cancer (CC) tissue. Methods From June 2014 to April 2017, 98 CC patients diagnosed and treated in the First Hospital of Handan were regarded as the research objects, and the surgically resected CC tissue and paracancerous tissue were collected. Methylation-specific PCR method was applied to determine the methylation status of FBN1 gene in CC tissue and paracancerous tissue. Western blot was applied to detection of FBN1 protein expression levels in CC tissue and paracancerous tissue. CC patients were followed up for 5 years and their survival conditions were recorded, the methylation rate of FBN1 gene in CC tissue and paracancerous tissue and the expression level of FBN1 protein in CC tissue of FBN1 gene non-methylation group and FBN1 gene methylation group were compared. The relationship between the methylation status of FBN1 gene in CC tissues and the clinicopathological characteristics of patients, the relationship between the methylation status of FBN1 gene and the prognosis of patients and the factors affecting the prognosis of CC patients were analyzed. Results The incidence of FBN1 gene methylation in CC tissues (60.20%) was higher than that in paracancerous tissues (12.24%) , and the difference was statistically significant (χ2=48.785, P<0.05). The expression level of FBN1 protein in FBN1 gene methylation group (0.61±0.12) was lower than that in FBN1 gene non-methylation group(1.59±0.32), and the difference was statistically significant(t=21.401, P<0.05). The methylation status of FBN1 gene in CC tissues was correlated with TNM staging, high risk-human papillomavirus DNA (HR-HPV DNA), and lymph node metastasis (χ2=7.578, 8.140, 7.814, all P<0.05). The 5-year cumulative survival rate of CC patients in the FBN1 gene methylation group (38.98%) was lower than that in the FBN1 gene non-methylation group (76.92%) , the difference was statistically significant (χ2=13.464, P<0.05).HR-HPV DNA positive [OR(95%CI): 2.534(1.577~4.072)], lymph node metastasis [OR(95%CI): 2.426(1.546~3.808)],TNM stageⅢ ~ Ⅳ [OR(95%CI): 2.702(1.633~4.471)] and FBN1 gene methylation [OR(95%CI): 2.394(1.531~3.743)] were independent risk factors affecting the prognosis of CC patients (P<0.05). Conclusion The level of FBN1 gene methylation in cancer tissues of CC patients was relatively high, which was related to clinicopathological characteristics such as TNM staging, HR-HPV DNA and lymph node metastasis, and prognosis, and it provides a new direction for clinical diagnosis and treatment of CC and evaluation of the prognosis of CC patients.