食管癌早期发现和高危人群预警液体活检技术分析

Analysis of liquid biopsy technique for early detection of esophageal cancer and early warning of high risk population

目的探讨液体活检技术对食管癌早期筛查和高危人群预警的意义。方法采用本实验室已经建立的间接酶联免疫吸附试验方法在早期食管癌患者和健康志愿者血清样本中检测3种肿瘤关抗原自身抗体细胞角蛋白19片段(CYFRA21-1)、增殖细胞核抗原(PCNA)和转化生长因子-β1(TGF-β1)的表达水平,分析这些指标联合检测对早期食管癌筛查的特异性和敏感性;采用最优的肿瘤相关抗原自身抗体组合进行小规模验证,在高发区无症状人群进行双盲液体活检和色素内镜活检病理,比较2种方法检测结果;采用该自身抗体技术在高发区无症状人群进行大规模食管癌筛查,了解检出情况。结果CYFRA21-1、PCNA和TGF-β1在早期食管癌中的阳性率分别为27.71%、31.67%、35.83%,均高于健康志愿者(2.92%、2.92%、5.00%),差异均有统计学意义(χ^2=113.752,P<0.001;χ^2=138.708,P<0.001;χ^2=140.425,P<0.001)。CYFRA21-1、PCNA和TGF-β1联合检测的敏感性67.08%,特异性76.04%,约登指数0.43。高发区无症状人群小规模验证结果证实,532例高发区无症状人群筛查发现阳性103例(19.36%),色素内镜检查结果示48例不典型增生患者、19例早期食管癌患者、9例中晚期食管癌患者、27例食管黏膜正常者;阴性429例(80.64%),色素内镜检查结果示11例不典型增生患者、2例早期食管癌患者、0例中晚期食管癌患者、416例食管黏膜正常者。高发区无症状人群大规模推广应用结果证实,10890例无症状人群经液体活检筛查发现,高危人群(阳性)1759例(16.51%),色素内镜检查结果示812例(12.22%)不典型增生患者、215例(46.16%)早期食管癌患者、23例(1.31%)中晚期食管癌患者、709例(40.31%)食管黏膜正常者。结论CYFRA21-1、PCNA及TGF-β1联合液体活检技术对食管癌早期发现及高危人群预警有重要意义,具有显著的推广应用价值。

Objective To investigate the significance of liquid biopsy for early cancer screening and early warning of high risk population in esophageal cancer.Methods We used the indirect enzyme-linked immunosorbent assay(ELISA)to respectively detect the expression levels of cell keratin 19 fragments(CYFRA21-1),proliferation cell nucleoants(PCNA)and transformational growth factors-β1(TGF-β1)in the patient and normal human serum samples,and analyzed the specificity and sensitivity of different combination indicators in screening early esophageal cancer.We used the optimal combination of tumor-related antigen autoantibodies for small-scale verification,while conducting a double-blind pigment endoscopic biopsy to compare the results of two methods of detection,and used this technique to screen for large-scale esophageal cancer in asymptomatic populations in high risk areas to understand the detection.Results The positive rates of CYFRA21-1,PCNA and TGF-β1 in the early esophageal cancer were 27.71%,31.67%and 35.83%,respectively,which were higher than those in the healthy volunteers(2.92%,2.92%,5.00%),and the differences were statistically significant(χ^2=113.752,P<0.001;χ^2=138.708,P<0.001;χ^2=140.425,P<0.001).The sensitivity,specificity and youden index of combined detection of CYFRA21-1,PCNA and TGF-β1 were 67.08%,76.04%and 0.43 respectively.The liquid biopsy results of small-scale validation of asymptomatic population in high incidence area confirmed that 103 patients(19.36%)were positive in 532 patients with asymptomatic population in high incidence area,the results of chromoendoscopy showed 48 patients with atypical hyperplasia,19 patients with early esophageal cancer,9 patients with advanced esophageal cancer and 27 patients with normal esophageal mucosa;429 patients(80.64%)were negative,and 11 patients with atypical hyperplasia were detected by chromoendoscopy,2 patients with early esophageal cancer,0 patient with advanced esophageal cancer and 416 patients with normal esophageal mucosa.The results of large-scale application of asymptomatic population in high risk areas confirmed that 1759(16.51%)of high risk population(positive)were found in 10890 asymptomatic population screening,812 patients(12.22%)with atypical hyperplasia,215 patients(46.16%)with early esophageal cancer,23 patients(1.31%)with advanced esophageal cancer,and 709 patients(40.31%)with normal esophageal mucosa.Conclusion CYFRA21-1,PCNA and TGF-β1 combined with liquid biopsy technology is of great significance for early detection of esophageal cancer and early warning of high risk population,and has significant promotional and application value.