外周血GP5、APC、GSTP1基因甲基化联合检测与乳腺癌相关性的研究

Correlation of GP5,APC and GSTP1 gene methylation with breast cancer

目的探索外周血GP5、APC、GSTP1基因甲基化联合检测与乳腺癌相关性。方法收集了2020年5月—2020年11月同济大学附属东方医院就诊的172例女性患者,其中有35例乳腺癌患者,90例乳腺良性疾病患者和47例其他组织来源的恶性肿瘤患者。实时荧光定量PCR法测定外周血中GP5、APC、GSTP1的基因甲基化表达水平,并与癌胚抗原(carcinoembryonic antigen,CEA)、糖类抗原153(carbohydrate antigen 153,CA153)、糖类抗原125(carbohydrate antigen 125,CA125)及乳腺钼靶进行比较。通过一致性检验和受试者工作特征(receiver operating characteristic,ROC)曲线分析并比较不同检测方法对乳腺癌的评价指标。分析三种基因联合检测的甲基化阳性率与乳腺癌临床病理特征相关性。结果35例乳腺癌患者和90例乳腺良性疾病患者外周血GP5、APC、GSTP1基因甲基化联合检测与组织病理结果的Kappa一致性为0.770。ROC曲线分析外周血基因甲基化联合检测的曲线下面积(area under the curve,AUC)为0.913(95%CI:0.8490.956),其中灵敏度为88.57%,特异度为92.22%,高于乳腺钼靶(AUC=0.765),高于血清CEA、CA125、CA153联合检测(AUC=0.761)。外周血基因甲基化与3种血清肿瘤指标联合检测的AUC最高为0.949(95%CI:0.8940.980)。35例乳腺癌患者外周血GP5、APC、GSTP1基因甲基化联合检测表达阳性率(88.6%)高于3种血清肿瘤标志物联合检测阳性率(28.6%);术前外周血基因甲基化表达的阳性率与年龄、淋巴结转移、肿瘤分期、肿瘤类型及脉管侵犯均无关(P>0.05)。47例其他组织来源恶性肿瘤与35例乳腺癌患者比较分析结果显示,外周血GP5、APC、GSTP1基因甲基化检测与病理的Kappa一致性为0.849。ROC曲线分析外周血基因甲基化检测的AUC为0.936(95%CI:0.8590.978),灵敏度为88.57%,特异度高达95.74%。结论外周血GP5、APC、GSTP1基因甲基化联合检测具有较高的灵敏度和特异度。外周血GP5、APC、GSTP1基因甲基化有望成为诊断乳腺癌的非侵入性生物标志物。

Objective To explore the correlation of methylation of GP5,APC and GSTP1 genes in peripheral blood with breast cancer.Methods A total of 172 female patients with breast disorders in Shanghai East Hospital Affiliated to Tongji University from May 2020 to November 2020 were enrolled,including 35 patients with breast cancer,90 patients with benign breast diseases,and 47 patients with malignant tumors derived from other tissues.The gene methylation levels of GP5,APC and GSTP1 in peripheral blood were determined by real-time PCR;the serum CEA,CA153 and CA125 levels were measured,and mammography was performed.The diagnostic value of different detection methods for breast cancer was analyzed by consistency test and receiver operating characteristic(ROC)curve.The correlation between the methylation of GP5,APC,GSTP1 genes and the clinicopathological characteristics of breast cancer was analyzed.Results The Kappa consistency between the GP5,APC,and GSTP1 gene methylation and the histopathological results was 0.770.ROC curve analysis showed that the area under the curve(AUC)of gene methylation was 0.913(95%CI:0.8490.956),with sensitivity of 88.57%and specificity of 92.22%.The AUC of mammography was 0.765 and that of combined detection of serum CEA,CA125 and CA153 was 0.761.The AUC of combination of gene methylation and serum tumor markers was the highest(0.949,95%CI:0.8940.980)(P<0.05).The positive rate of GP5,APC,GSTP1 gene methylation was higher than that of combined detection of serum tumor markers(88.6%vs.28.6%).The positive gene methylation in breast cancer was not correlated with age,lymph node metastasis,tumor stage,tumor type and vascular invasion(P>0.05).In 47 cases of other tissue-derived malignant tumors and 35 cases of breast cancer,the Kappa consistency between GP5,APC,GSTP1 gene methylation and pathological diagnosis was 0.849;ROC aurve analysis showed that the AUC of peripheral blood gene methylation detection was 0.936(95%Cl 0.8590.978),the sensitivity was 88.57%and the specificity was 95.74%.Conclusion The methylation detection of GP5,APC and GSTP1 genes in peripheral blood has high sensitivity and specificity in diagnosis of breast cancer,indicating that it would be a non-invasive biomarker for diagnosing breast cancer and deserves further exploration.