半胱氨酸蛋白酶抑制剂S与传统肿瘤标志物在胃肠癌中的诊断价值比较.pdf

实验与检验医学 2 0 2 3 年8 月第41卷第4期 Experimental and Laboratory Medicine,Aug.2023,Vol.41,No.4半胱氨酸蛋白酶抑制剂S与传统肿瘤标志物在胃肠癌中的诊断价值比较周爱群,张志勇，方全钢，曾黎峰,刘燕鸣,孔维莉（江西省人民医院（南昌医学院第一附属医院）检验科，江西南昌330 0 0 6)摘要：目的探讨血清半胱氨酸蛋白酶抑制剂S(CST4)在胃肠癌中的临床意义。方法利用血清CST4试剂盒(ELISA法)和化学发光法分别检测8 0 例胃肠癌病人（胃癌40 例,结直肠癌40 例)和40 例正常人对照血清CST4、C E A、C A 19-9 和CA125浓度，并对检测结果进行统计分析，评估CST4的临床意义。结果胃肠癌组CST4浓度中位数分别为7 8.2 7 2(36.8 9 2，188.395）U/L 和6 6.2 6 5（39.7 51,10 8.8 9 0）U/L，均显著高于正常对照组中位数30.9 2 0（16.0 55,56.9 9 5)U/L，差异有统计学意义(P0.05)。胃肠癌患者CST4的ROC曲线下面积(AUC)分别为0.7 55、0.7 54)、胃肠癌早期（一期和二期)CST4的阳性检出率（均为2 6.9%)和检测灵敏度、特异性、阳性似然比(PLR）阴性似然比(NLR）、阳性预测值(PPV)和阴性预测值（NNV)等检测指标均优于CEA，C A 19-9 和CA125。胃肠癌+I V 期患者CST4中位数水平分别为16 5.6 53(58.7 6 7,334.9 33）U/L和7 4.36 5（41.19 2,18 3.30 6）U/L,均高于胃肠癌I期和期患者，胃癌组水平差异有统计学意义(P0.05）。结论胃肠癌组血清CST4水平显著高于正常对照组，早期阳性检出率和诊断性能优于传统肿瘤标志物，并且与肿瘤预后相关，有望成为新的胃肠癌早期肿瘤标志物。关键词：半胱氨酸蛋白酶抑制剂S；胃肠癌；肿瘤标志物中图分类号：R735文献标识码：A文章编号：16 7 4-112 9(2 0 2 3)0 4-0 38 3-0 5D0I:10.3969/j.issn.1674-1129.2023.04.002The diagnostic value comparison of CST4 and three traditional biomarkers in gastrointestinal cancers ZHOU Aiqun,ZHANG Zhiyong,FANG Quangang,et al Jiangxi Provincial Peoples Hospital,The First AFfiliation Hospital of Nanchang MedicalColleye,Jiangxi Nanchang,330006,China.Abstract:Objective To explore the clinical applications of cystatin S(CST4)in gastrointestinal cancers.Methods A total of80 serum samples from gastrointestinal cancer patients including 40 gastric cancer(GC)and 40 colorectal cancer(CRC)Jand 40serum samples from healthy controls were all detected by human CST4 detective kit(ELISA)and CEA,CA19-9 and CA125 kits(CLA),then Statistical analysis were made to evaluate the clinical values of CST4 in gastrointestinal cancers.Results The medianconcentrations of CST4 in GC group and CRC group 78.272(36.892,188.395)U/L and 66.265(39.751,108.890)U/L respective-ly are significantly higher than those of normal controls30.920(16.055,56.995)U/LJ(P0.05).The area under the R0C curve(AUC)of CST4 for GC and CRC(0.755 and 0.754 respectively),the positive detection rate in the early stages(stage I and II)ofGC and CRC(both 26.9%)and the diagnostic sensitivity,specificity,positive likelihood ratio(PLR),negative likelihood ratio(NLR),positive predictive value(PPV)and negative predictive value(NNV)of CST4 are all superior to CEA,CA19-9 and CA125.The median concentrations of CST4 in stages II+IV of GC and CRC group 165.653(58.767,334.933)U/L and 74.365(41.192,183.306)U/L respectively are both higher than those of stage I and stageII,and the difference of GC group is statistically signifi-cant as well(P0.05)。表1各组血清样本临床病例资料统计表分组分期胃癌1期期期IV期总数结直肠癌总数正常对照1.2主要试剂人半胱氨酸蛋白酶抑制剂S(CST4)检测试剂盒（酶联免疫法）,上海良润生物医药科技有限公司生产。CEA、CA 19-9、CA 12 5化学发光法检测试剂（罗氏生产）。实验与检验医学2 0 2 3年8 月第41卷第4期Experimental and LaboratoryMedicine，A u g.2 0 2 3,Vo l.41,No.41.3主要仪器全自动酶标仪（BioTekElx800）、全自动酶标洗板机(PW-960）。全自动化学发光免疫分析仪(罗氏E601)。1.4 研究方法1.4.1血清CST4的检测严格按照试剂盒说明书操作。从4冰箱取出所需人份的人半胱氨酸蛋白酶抑制剂S(CST4)检测试剂盒（酶联免疫法）,平衡至室温；将待检血清从-8 0 冰箱取出解冻平衡至室温待检。每板设置7 个浓度校准品及1个质控孔，加入10 0 l校准品和质控品，再依次加入10 0L待检血清,37 水浴箱孵育1h;全自动洗板机洗板5次后每孔加人10 0 L酶标抗体，37 水浴箱孵育1h；全自动洗板机洗板5次后每孔加入配置好的10 0 l底物液，37 水浴箱避光显色15min,最后加人10 0 l终止液终止反应。利用酶标仪15min内完成比色测定各孔OD值,绘制标准曲线并计算各个样本浓度值。1.4.2血清CEA、CA 19-9、CA 12 5浓度检测按照全自动化学发光免疫分析仪的标准操作流程检测。1.4.3绘制受试者工作特征(ROC)曲线评价CST4在胃肠癌中的诊断价值。1.4.4统计学处理全部资料采用SPSS17.0统计软件包进行统计分析。呈正态分布的计量资料以x士s表示，非正态分布的计量资料用M(P25,P75)表示；正态性连续资料用单因素方差分析，两两比较用最小显著性差异法分析。非正态性连续资料用非参数检验。P0.05为差异有统计学意义。n男女比年龄范围188113401期16期10期12IV期240402.1各组血清CST4测定结果采用SPSS17.0统计软件对各组血清CST4测定结果进行夏皮洛-威尔克(S-W)检验和直方图分析，结果发现数据不服29/11368428/12308224/162980从正态分布，故采用两样本Mann-WhitneyU检验,结果见表2。表2 胃癌组、结直肠癌组和正常对照组血清 CST4浓度比较(U/L)分组n胃癌组40结直肠癌组40正常对照组40注：*与正常对照组比较，CST4浓度中位数分布差异存在统计学意义,P0.001。2.2CST4对胃肠癌的早期诊断价值按试剂盒说明书参考范围确定各指标的阴阳性结果，分别计中位数四分位数1四分位数278.272*36.89266.265*39.75130.92016.055188.395108.89056.995实验与检验医学2 0 2 3年8 月第41卷第4期Experimental and Laboratory Medicine,Aug.2023,Vol.41,No.4算各指标在胃肠癌早期（I期和期)中的阳性检出率,结果发现CST4在胃肠癌早期中的阳性检出率均达2 6.9%,高于传统肿瘤标志物CEA、CA 19-9和CA125,结果见表3。表3胃肠癌早期(I期和期）各检测指标阳性检出率比较分组CST4胃癌组26.9%(7)结直肠癌组26.9%(7)2.3 CST4 在胃肠癌中的诊断价值及与传统肿瘤标志物的比较利用SPSS17.0统计软件绘制各指标受试者工作特征(ROC)曲线。结果显示,CST4、CE A、CA19-9、CA 12 5对胃癌预测的AUC分别为0.7 55(95%CI:0.6490.861,P0.001)0.628(95%CI:0.5050.750,P0.05)、0.6 47(9 5%C I:0.52 4 0.7 2 0,P 0.0 5);对结直肠癌预测的ROC曲线下面积（AUC）分别为0.7 54(95%CI:0.6500.859,P0.001)、0.6 6 6 (9 5%C I:0.5490.784,P0.05),0.661(95%CI:0.5400.781,P 0.0 5)。可见,CST4对胃肠癌预测的AUC 均高于传统肿瘤标志物，诊断价值优于传统肿瘤标志物。结果见图1。2.3 CST4 在胃肠癌的诊断性能评估及与传统肿瘤标志物比较胃肠癌组所有病例以病理诊断结果为金标准,分别计算各指标检测临床灵敏度、特异性、PLR、NL R、PPV和NNV。结果发现,CST4的所有检测性能指标均优于 CEA、CA 19-9 和 CA125。利用SPSS17.0统计软件包对CST4和传统肿瘤标志物的灵敏度和特异性进行配对检验，发现检测指标灵敏度CST435.0%CEA17.5%CA19-922.5%CA12512.5%注：与CA125比较，灵敏度差异有统计学意义,=4.9 2 3,P0.05。表5血清CST4对结直肠癌诊断性能及与传统肿瘤标志物的比较检测指标灵敏度CST430.0%*CEA17.5%CA19-920.0%CA1255.00%注：与CA125比较,灵敏度差异有统计学意义,=8.1,P0.05。385.AB1.0CST4CEACA10-90.8CA125梦专线阳性检出率0.2CEACA19-97.7%(2)15.4%(4)11.5%(3)19.2%(5)表4血清CST4对胃癌诊断性能及与传统肿瘤标志物的比较特异性97.5%95.0%92.5%95.0%特异性97.5%95.0%92.5%95.0%1.00.80.6-0.4-0.2CA1250.0十0.011.5%(3)3.8%(1)PLR143.532.5PLR123.52.6671CST4CEACAI9-9CA1250204060.80.01.00.01-特异性图1胃癌(A）及结直肠癌(B）血清肿瘤标志物受试者工作特征曲线（ROC）分析注：*P0.05;*P0.001。CST4检测灵敏度与CA125比较差异有统计学意义（胃肠癌组分别为=4.9 2 3,P0.05和x2=8.1,P0.05），结果分别见表4和表5。2.4CST4对胃肠癌预后价值的评估为了评估CST4对胃肠癌预后判断价值，我们比较了胃肠癌晚期（+IV期)和早期(I期和期)CST4的水平，采用Kruskal-WallisH检验，结果发现胃肠癌+IV期CST4中位数分布水平均高于胃肠癌I期和期，其中胃癌+I V期CST4水平与胃癌I期和I期比较差异有统计学意义(2=7.8 17,P0.05)。结果分别见图2 和表6。3讨论半胱氨酸蛋白酶抑制剂(Cystatins,CSTs)是一类对半胱氨酸蛋白酶具有抑制作用的蛋白家族 9-10 ,越来越多的研究发现CSTs与肿瘤的发生发展密切相关,CSTs 在肿瘤的生长-12 、血管生成 13、浸润 14和转移 15 中起重要作用，具有作为肿瘤诊断和预后评估标志物的潜力。NLR0.6670.8680.8370.921NLR0.7180.8680.86412040.60.81.01-特异性PPVNNV93.3%60.0%77.8%53.5%75.0%54.4%71.4%52.0%PPVNNV92.3%58.2%77.8%53.5%72.7%53.6%50.0%50.0%表6 胃肠癌不同分期CST4水平分布(U/L）比较386分组胃癌结直肠癌注：与胃癌I期和I期比较，中位数分布差异有统计学意义,X2=7.817,P0.05。60:000-500:0300.00(UIL)200.001000.00实验与检验医学2 0 2 3年8 月第41卷第4期Experimental and LaboratoryMedicine，A u g.2 0 2 3,Vo l.41,No.4分期I期I期II+IV期I期期II+IV期600.00800.000CST4400.00900.00(UL)200.000100.00中位数59.432132.005165.653*62.98158.00774.365工四分位数126.96439.23758.76734.03237.08441.192参考文献1JSung H,Ferlay J,Siegel RL,et al.,Global Cancer Statistics 2020:CLOBOCAN Estimates of Incidence and Mortality Worldwide for36 Cancers in 185 Countries.J.CA Cancer J Clin 2021;71:209-249四分位数288.094210.278334.933101.751115.072183.306n18814161014分期2Ferlay J,Colombet M,Soerjomataram I,et al.Cancer statistics for图2 胃肠癌不同分期CST4水平(U/L)分布比较胃肠癌传统肿瘤标志物有 CEAl0、CA 19-9 l 17-18 、CA12519等,但是这些指标的灵敏度和特异性都不够理想。本研究发现胃肠癌组血清CST4水平显著高于正常对照组，且差异具有统计学意义，提示CST4可作为一项新的胃肠癌肿瘤标志物。我们比较了各指标在胃肠癌早期（I期和期)中的阳性检出率，结果发现CST4早期阳性检出率均高于CEA、C A 19-9 和CA125,提示CST4有望用于胃肠癌的早期筛查，有利于胃肠癌的早期诊断和早期治疗。通过绘制ROC曲线分析，我们发现胃肠癌组CST4 的 AUC 均高于传统肿瘤标志物,结果与学者吴明兰等 2 0 和石梦珍等 2 1 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