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中华肺部疾病杂志(电子版) ›› 2024, Vol. 17 ›› Issue (04) : 505 -511. doi: 10.3877/cma.j.issn.1674-6902.2024.04.001

论著

肺泡灌洗液RASSF1A及SHOX2甲基化联合径向超声特征对肺结节性质鉴别诊断的意义
刘锦程1, 王斌1, 张雯1, 张明周1, 刘禹1, 叶东樊1, 黄赞胜1, 邱凌霄1, 卿斌1, 王创业1, 王南博1, 王苹1, 郭宇航1, 周培花1, 程秋霞1, 徐智1,†()   
  1. 1. 400037　重庆，陆军（第三）军医大学第二附属医院呼吸与危重症医学中心；400037　重庆，重大呼吸疾病精准诊疗与防控重庆市重点实验室
  • 收稿日期:2024-06-18 出版日期:2024-08-25
  • 通信作者: 徐智
  • 基金资助:
    重庆市技术创新与应用发展专项重点项目(CSTC2021jscx-gksb-N0029)

Significance of RASSF1A and SHOX2 methylation in bronchoalveolar lavage fluid combined with radial ultrasonic features in differentiating the property of pulmonary nodules

Jincheng Liu1, Bin Wang1, Wen Zhang1, Mingzhou Zhang1, Yu Liu1, Dongfan Ye1, Zansheng Huang1, Lingxiao Qiu1, Bin Qing1, Chuangye Wang1, Nanbo Wang1, Ping Wang1, Yuhang Guo1, Peihua Zhou1, Qiuxia Cheng1, Zhi Xu1,†()   

  1. 1. Department of Respiratory and Critical Care Medical Center, The Second Affiliated Hospital of Army Medical University, Chongqing 400037, China; Chongqing Key Laboratory for Precise Diagnosis, Treatment, Prevention and Control of Major Respiratory Diseases, Chongqing 400037, China
  • Received:2024-06-18 Published:2024-08-25
  • Corresponding author: Zhi Xu
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引用本文：

刘锦程, 王斌, 张雯, 张明周, 刘禹, 叶东樊, 黄赞胜, 邱凌霄, 卿斌, 王创业, 王南博, 王苹, 郭宇航, 周培花, 程秋霞, 徐智. 肺泡灌洗液RASSF1A及SHOX2甲基化联合径向超声特征对肺结节性质鉴别诊断的意义[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 505-511.

Jincheng Liu, Bin Wang, Wen Zhang, Mingzhou Zhang, Yu Liu, Dongfan Ye, Zansheng Huang, Lingxiao Qiu, Bin Qing, Chuangye Wang, Nanbo Wang, Ping Wang, Yuhang Guo, Peihua Zhou, Qiuxia Cheng, Zhi Xu. Significance of RASSF1A and SHOX2 methylation in bronchoalveolar lavage fluid combined with radial ultrasonic features in differentiating the property of pulmonary nodules[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 505-511.

目的

分析检测肺泡灌洗液中大鼠肉瘤(rat sarcoma, Ras)相关区域家族1A基因(Ras association family 1 isoform A, RASSF1A)和矮小同源盒基因2(short stature homeobox 2, SHOX2)甲基化联合径向超声特征对肺结节性质鉴别诊断的意义。

方法

选择2022年9月至2024年3月我院收治的肺结节患者90例，经组织病理学确诊恶性肺结节54例为观察组，良性肺结节36例为对照组，收集肺泡灌洗液进行细胞学检查及RASSF1A、SHOX2基因甲基化检测，比较两组甲基化阳性率，分析肺泡灌洗液甲基化检测对肺结节性质的灵敏度和特异性，采用Logistic回归分析从临床特征、径向超声特征及甲基化指标中筛选预测因子，构建恶性肺结节预测模型。

结果

观察组RASSF1A阳性22例(40.74%)高于对照组1例(2.78%)(P<0.01)、SHOX2阳性25例(46.30%)高于对照组2例(5.56%)(P<0.01)、RASSF1A及SHOX2两者阳性14例(25.93%)高于对照组1例(2.78%)(P<0.05)、RASSF1A或SHOX2阳性33例(61.11%)高于对照组2例(5.56%)(P<0.01)；RASSF1A或SHOX2阳性诊断恶性肺结节的灵敏度61.11%、特异性94.44%。RASSF1A或SHOX2阳性33例(61.11%)高于肺泡灌洗液细胞学阳性17例(31.48%)(P<0.05)；RASSF1A或SHOX2阳性联合肺泡灌洗液细胞学阳性率72.22%；观察组径向超声特征中边缘连续比率81.40%、内部回声不均匀比率83.72%及低回声影比率72.09%高于对照组40.00%、40.00%、45.71%(P<0.05)；多因素Logistic回归分析显示，两基因甲基化、径向超声特征边缘连续及内部回声不均匀为预测恶性肺结节的危险因素，绘制受试者工作特征曲线下面积(area under the curve, AUC)为0.886。

结论

RASSF1A联合SHOX2甲基化检测肺泡灌洗液用于恶性肺结节诊断具有价值，RASSF1A及SHOX2甲基化联合径向超声特征对肺结节性质鉴别诊断有临床意义。

关键词: 肺结节, 肺泡灌洗液, 大鼠肉瘤相关区域家族1A基因甲基化, 矮小同源盒基因甲基化, 径向超声特征
Objective

To analyse the significance of methylation test of Ras association family 1 isoform A (RASSF1A) and short stature homeobox 2 (SHOX2) in bronchoalveolar lavage fluid(BALF) combined with radial ultrasonic features in the differential diagnosis between malignant and benign pulmonary nodules.

Methods

Ninety patients with pulmonary nodules admitted to our hospital from September 2022 to March 2024 were selected, 54 cases of malignant pulmonary nodules diagnosed by histopathology were in the observation group, and 36 cases of benign pulmonary nodules were in the control group, and BALF was collected for cytological examination and methylation detection of the RASSF1A and SHOX2 genes to compare the methylation positivity rate of the two groups and analyse the sensitivity and specificity of pulmonary nodules. Logistic regression analysis was used to screen some clinical characteristics, radial ultrasonic features and methylation indexes as predictors to construct a prediction model of pulmonary nodules.

Results

In the observation group, 22 cases (40.74%) of RASSF1A positivity were higher than 1 case (2.78%) in the control group(P<0.01); 25 cases (46.30%) of SHOX2 positivity were higher than 2 cases (5.56%) in the control group(P<0.01); 14 cases (25.93%) of both RASSF1A and SHOX2 positivity were higher than 1 case(2.78%) in the control group(P<0.05); 33 cases (61.11%) of RASSF1A or SHOX2 positivity were higher than 2 cases (5.56%) in the control group(P<0.01; The sensitivity for diagnosing malignant lung nodules was 61.11% and the specificity was 94.44% for the positivity of RASSF1A or SHOX2. 33 cases (61.11%) of RASSF1A or SHOX2 positivity were significantly higher than 17 cases(31.48%) of BALF cytology positivity(P<0.05); Radial ultrasound characteristics of the observation group in the ratio of the continuous margins of 81.40%, the ratio of the uneven internal echo of 83.72% and the ratio of hypoechogenic shadows of 72.09% were higher than that of the control group of 40.00%, 40.00%, 45.71% (P<0.05); Multifactorial logistic regression analysis showed that two-gene methylation, continuous margins and uneven internal echo were the risk factors for predicting malignant lung nodules, and the area under the curve (AUC) of the working characteristics of the subjects was 0.886.

Conclusion

RASSF1A combined with SHOX2 methylation detection of BALF is valuable for the diagnosis of malignant lung nodules, and RASSF1A and SHOX2 methylation combined with radial ultrasound features are clinically relevant for the differential diagnosis of the property of lung nodules.

Key words: Pulmonary nodule, Bronchoalveolar lavage fluid, Ras association family 1 isoform A methylation, Short stature homeobox 2 methylation, Radial ultrasound features
表1 RASSF1A及SHOX2对肺结节的诊断[n(%)]
甲基化指标   组织病理学 合计(n＝90) 灵敏度 特异性 PPV NPV
恶性(n＝54) 良性(n＝36)
RASSF1A 阳性 22 1 23 40.74 97.22 95.65 52.24
  阴性 32 35 67        
SHOX2 阳性 25 2 27 46.30 94.44 92.59 53.97
  阴性 29 34 63        
RASSF1A及SHOX2 阳性 14 1 15 25.93 97.22 93.33 46.67
  阴性 40 35 75        
RASSF1A或SHOX2 阳性 33 2 35 61.11 94.44 94.29 61.82
  阴性 21 34 55        
表2 肺泡灌洗液甲基化不同病理类型比较[n(%)]
病理类型 例数 RASSF1A阳性 SHOX2阳性 RASSF1A及SHOX2阳性 RASSF1A或SHOX2阳性
肺腺癌 33 11(33.33) 8(24.24) 3(9.09) 16(48.48)
肺鳞癌 13 4(30.76) 10(76.92) 4(30.76) 10(76.92)
小细胞肺癌 6 6(100.00) 6(100.00) 6(100.00) 6(100.00)
未明确类型 2 1(50.00) 1(50.00) 1(50.00) 1(50.00)
统计量   10.082 18.633 20.756 7.557
P值   0.009 0.000 0.000 0.031
表3 肺泡灌洗液甲基化不同临床分期比较[n(%)]
临床分期 例数 RASSF1A阳性 SHOX2阳性 RASSF1A及SHOX2阳性 RASSF1A或SHOX2阳性
Ⅰ＋Ⅱ 26 8(30.80) 11(42.30) 6(23.10) 13(50.00)
Ⅲ＋Ⅳ 24 10(41.70) 11(45.80) 5(20.80) 16(66.70)
未明确分期 4 4(100.00) 3(75.00) 3(75.00) 4(100.00)
Fisher确切法   6.414 1.440 4.626 3.804
P值   0.039 0.596 0.099 0.130
表4 甲基化与肺泡灌洗液细胞学对肺结节诊断的比较[n(%)]
方法学及叠加 灵敏度 特异性 P值
BALF细胞学阳性 17(31.48) 36(100.00)  
RASSF1A阳性 22(40.74) 35(97.22) 0.809
SHOX2阳性 25(46.30) 34(94.44) 0.063
RASSF1A及SHOX2阳性 14(25.93) 35(97.22) 0.159
RASSF1A或SHOX2阳性 33(61.11) 34(94.44) 0.004
BALF细胞学或RASSF1A或SHOX2阳性 39(72.22) 34(94.44) 0.000
图1 支气管径向超声特征分类。注：A：边缘连续清晰(腔内型)；B：病灶与探头关系(腔外型)；C：支气管充气征、低回声影；D：内部回声不均
图2 甲基化与径向超声特征预测肺结节良恶性。注：RASSF1A：大鼠肉瘤相关区域家族1A基因；SHOX2：矮小同源盒基因2；RASSF1A＋SHOX2：RASSF1A阳性或SHOX2阳性；径向超声特征：边缘连续和内部回声不均匀；多指标联合：RASSF1A＋SHOX2与径向超声特征联合
表5 单因素及多因素Logistic回归筛选恶性肺结节预测模型
变　量 单因素Logistic回归 多因素Logistic回归
β Z OR(95%CI) P值 β Z OR(95%CI) P值
RASSF1A联合SHOX2 3.285 17.770 26.714(5.799～123.055) 0.000 2.697 9.594 14.837(2.692～81.766) 0.002
边缘连续 1.881 12.984 6.562(2.358～18.260) 0.000 1.399 4.649 4.052(1.136～14.454) 0.031
内部回声不均 2.043 14.409 7.714(2.686～22.153) 0.000 1.613 4.864 5.019(1.197～21.053) 0.027
低回声影 1.121 5.446 3.068(1.197～7.865) 0.020 －0.088 0.015 0.916(0.230～3.645) 0.901
吸烟史 0.947 4.591 2.579(1.084～6.135) 0.032 0.658 1.143 1.930(0.578～6.444) 0.285
支气管充气征 0.480 1.067 1.615(0.650～4.012) 0.302 －2.592常量 12.144 0.075 0.000
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