胆囊癌mAb检测消化道肿瘤血清癌相关抗原
浙江医科大学附属二院外科 浙江省杭州市 310009
梁刚，男，1964-08-23生，浙江省杭州市人，汉族.1987年浙江医科大学医学系本科毕业；外科学博士.主要从事胆囊癌的诊治研究，发表论文7篇.
项目负责人 梁刚，浙江省杭州市解放路68号.
Tel: 0571-7077272-2602
收搞日期 1996-06-15 接受日期 1996-07-12
Measurement of serum carcino-associated antigen by anti-gallbladder mAb in digestive tract cancer
, 百拇医药
Gang Liang, Yuan-Ming Wang, Shu-You Peng, Zheng-Rong Shen and Fan Zhou
Department of Surgery, the Second Affiliated Hospital of Zhejiang Medical University, Hangzhou 310009, Zhejiang Province, China
Abstract
AIM To diagnose digestive tract cancer serologically using anti-gallbladder mAb.
METHODS Gallbladder associated antigen (GBAA) levels were measured by mAb for gallbladder cancer and conA in patients with digestive tract cancer, benign diseases and healthy controls. Serum CEA, AFP, CA19-9 and CA 242 levels were also measured simultaneously in the subjects.
, 百拇医药
RESULTS The GBAA positive rates were as follows: 2.2% in normal controls, 13.3% in benign diseases, 92.3% in gallbladder cancer, 20.9% in hepatocelullar cancer, 9.1% in pancreas cancer, 50% in gastric cancer, 41.6% in colon cancer, 50% in bile duct cancer, 100% in cancer of the ampulla of Vater. In gallbladder cancer, hepatocelullar cancer, pancreas cancer, gastric cancer, colon cancer, bile duct cancer, ampulla of Vater cancer, CEA positive rates were 7.6%, 3.8%, 33.3%, 21.5%, 33.3%, 0% and 0% respectively, AFP positive rates 0%, 57%, 6%, 0%, 0% and 0%, respectively, CA19-9 positive rates 62%, 3.8%, 77.7%, 0%, 50%, 0% and 50%, respectively, and CA242 positive rates 62%, 0%, 66.6%, 0%, 50%, 0% and 25% respectively. The sensitivity and specificity in diagnosis of gallbladder cancer by GBAA were 92.3% and 94.1% respectively. The sensitivity in diagnosis of tumors of non-gallbladder cancer was 32%. GBAA was also valuable for post-operative or recurrence monitoring. No correlation was found between the GBAA and tumor sizes, infiltration, differentiation and metastasis.
, 百拇医药
CONCLUSION Serum GBAA is useful for diagnosing gallbladder cancer, without cross reaction between GBAA and CEA or AFP.
Subject headings Gallbladder neoplasms Digestive system neoplasms Antibody, monoclonal CA_19_9 antigen/analysis Carcinoembryonic antigen/analysis α-fetoproteins/analysis
Liang G, Wang YM, Peng SY, Shen ZR, Zhou F. Measurement of serum carcino-associated antigen by anti-gallbladder mAb in digestive tract cancerXin Xiaohuabingxue Zazhi, 1997;5(5):323-324
, 百拇医药
摘 要目的 应用胆囊癌mAb对消化道肿瘤患者进行血清学诊断.
方法 采用胆囊癌mAb及刀豆蛋白A检测206例各种消化道肿瘤(胆囊癌13例、肝癌39例、胃癌39例、胰腺癌15例、胆管癌5例、壶腹部癌6例，结肠癌14例).良性疾病(30例)和健康供血者(45例)血清中胆囊癌相关抗原(GBAA)进行测定；同时测定CEA，AFP，CA19-9及CA242含量.
结果 正常人血清GBAA阳性率2.2%，良性疾病13.3%，胆囊癌GBAA 92.3%，CEA7.6%，AFP 0%，CA19-9 62%,CA242 62%;肝癌GBAA 20.9%，CEA3.8%，AFP 57.6%，CA19-9 3.8%,CA242 0%;胰腺癌GBAA 9.1%，CEA 33.3%，AFP 0%，CA19-9 77.7%,CA242 66.6%;胃癌GBAA 50%，CEA 21.5%，AFP 6%，CA19-9 0%,CA242 0%;大肠癌GBAA 41.6%，CEA 33.3%，AFP 0%，CA19-9 50%,CA242 50%;胆管癌GBAA50%，CEA 0%， AFP 0%，CA19-9 0%，CA242 0%；壶腹部癌GBAA 100%，CEA 0%，AFP 0%，CA19-9 50%,CA242 25%. GBAA对胆囊癌诊断敏感性92.3%，特异性94.1%，对非胆囊癌肿瘤诊断敏感性32%.GBAA对肿瘤术后及复发监测有一定价值.GBAA与胆囊癌肿瘤大小，浸润程度、组织分化及有无转移无相关性.
, http://www.100md.com
结论 GBAA单抗术前对胆囊癌诊断较高；GBAA单抗与CEA、AFP无交叉反应；临床上测定GBAA可作为CEA，AFP，CA19-9及CA242的补充，为临床诊断及普查提供了新的肿瘤标记物.
主题词 胆囊肿瘤 消化系统肿瘤 抗体，单克隆 CA-19-9抗原/分析 癌胚抗原/分析 甲胎蛋白/分析
梁刚, 王远明, 彭淑牖, 沈正荣, 周凡.胆囊癌mAb检测消化道肿瘤血清癌相关抗原.新消化病学杂志,1997;5(5):323-324
应用肿瘤单克隆抗体(monoclonal antibody, mAb)对恶性肿瘤患者进行血清学诊断是当今临床重点研究课题之一.中晚期胆囊癌生存率低，若早期诊断，可提高手术切除率，延长患者生命.作者自制胆囊癌mAb，建立ELISA法并对消化道肿瘤、良性疾病及健康供血者血清中胆囊癌相关抗原(GBAA)测定.同时对消化道肿瘤患者测定CEA，AFP，CA19-9，CA242，比 较其敏感性和特异性.
, http://www.100md.com
1 对象和方法
1.1 对象
抗体：以胆囊癌细胞作抗原，免疫Balb/c小鼠，与NS-1骨髓瘤细胞融合，获得分泌抗胆囊癌抗原(GBAA)的单克隆抗体.辣根过氧化酶(HRP.Sigma).CNBr-Sepharose 4B(Pharmacia).刀豆蛋白A(con A, Sigma).CEA试剂盒，AFP试剂盒(Syntron USA),CA19-9试剂盒(Sorin)，CA242试剂盒(Pharmacia). 血清标本：共206例，女性70例，男136例，平均年龄41岁. 其中胆囊癌13例，肝癌39例，胃癌39例，胰腺癌15例，胆管癌5例，壶腹部癌6例，结肠癌14例.良性疾病30例，其中慢性胆囊炎17例，急性胰腺炎8例，胃溃疡2例，肝脓肿2例，肝血管瘤1例，正常人45例.空腹抽血经离心分离血清，编号-20℃保存.
, 百拇医药
1.2 方法 GBAA测定：用150mg/L Con A包被微孔，4℃过夜，次日取出，洗涤3次，加入待检血清4℃反应6h，洗涤3次，加入纯化GBAA抗原标记的HRP，37℃ 1h，洗涤3次显 色，测各孔OD值.CEA，AFP，CA19-9，CA242均采用酶免疫法测定，按试剂说明书操作.统计学处理 采用SPSS统计软件包，卡方检验.Fisher's exact test.
2 结果GBAA OD值为正常人的x±2s判为阳性；正常值CEA≤2.5μg/L，AFP≤20μg/L， CA19-9≤37Ku/L，CA242≤17Ku/L.消化道肿瘤血清中肿瘤标志物表达阳性率见表1.
表1 消化道肿瘤血清中肿瘤标志物表达阳性率 (%)
, 百拇医药
非肿瘤患者和人群血清中GBAA表达阳性率，正常人为1/45，良性疾病为4/30，假阳性6.7%，特异性93.3%；其中胆囊炎阳性率为1/11，与胆囊癌比较12/13，差别非常显著(P<0.01)，敏感性92.3%，特异性94.1%；非胆囊癌肿瘤的阳性率为32/100，差别也很显著(P<0.01). 研究发现术前GBAA阳性，术后切除的GBAA转阴31例；如不能切除则GBAA仍然阳性13例.如肿瘤复发则GBAA转阳性8例.GBAA与胆囊癌肿瘤大小、浸润程度、组织分化及淋巴转移无相关.
3 讨论胆囊癌居消化道肿瘤第6位，且近年有上升趋势.晚期胆囊癌预后极差，欲提高患者的预后，只有早期诊断，早期治疗胆囊癌.CEA，CA19-9及CA242术前诊断胆囊癌已有不少报道[1，2]，但缺乏敏感性和特异性.本研究发现GBAA在胆囊癌、胃癌、肝癌、胰腺癌、大肠癌、胆管癌及壶腹部癌阳性率分别为92.3%，50.0%，20.9%，9.1%、41.6%，50.0%及100%.GBAA对胆囊癌的敏感性92.3%，特异性94.1%，GBAA对临床诊断胆囊癌有较大价值. GBAA假阳性为6.7%.正常人群中出现少数假阳性，其原因有待进一步探讨.同时发现GBAA对胆管癌、壶腹癌肿瘤诊断有一定价值，由于例数少，有待于进一步研究.GBAA结合CA19-9或CA242有助于胆囊癌与胰腺癌之间的鉴别诊断.可见抗胆囊癌单克隆抗体可作为CEA，CA19-9的补充，为开展对胆囊癌的普查、临床诊断提供了一种新的肿瘤标志物.各种肿瘤 如术前GBAA阳性，手术切除后均转阴性，如肿瘤未切除，则GBAA持续阳性.发现术后复发(特别是胆囊癌患者)，则GBAA阳性.化疗后，GBAA测定为阴性，但观察例数不多，GBAA能否作为化疗后疗效判断需要进一步研究.肿瘤标志物应能指示出肿瘤大小、浸润程度、有无转移及病情的发展情况[3].本文研究GBAA与胆囊癌的肿瘤大小、组织类型、浸润程度及淋巴结转移关系无相关性.分析原因可能有：①病例数少，统计学处理无差异；②手术记录数字不精确；③肉眼无法判断有无微小转移灶.
, http://www.100md.com
4 参考文献1 Brian L, Maislin G, Suzanne L. Serum CEA and CA19-9: potential future diagnostic or screening tests for gallbladder cancer.
Int J Cancer, 1990;45(12):821-824
2 Ritts H, Nayorney M, Jacobsen G. Comparison of preoperative serum CA19-9 levels with results of diagnostic imaging modalities
in patients undergoing laparotomy for suspected pancreatic or gallbladder disease.Pancreas, 1994;9(6):707-716
3 Markus A, Helen J, Felix L. CA242 is a new tumor marker for pancreatic cancer. Cancer,1993;71(3):701-707, http://www.100md.com(梁刚 王远明 彭淑牖 沈正荣 周凡)
梁刚，男，1964-08-23生，浙江省杭州市人，汉族.1987年浙江医科大学医学系本科毕业；外科学博士.主要从事胆囊癌的诊治研究，发表论文7篇.
项目负责人 梁刚，浙江省杭州市解放路68号.
Tel: 0571-7077272-2602
收搞日期 1996-06-15 接受日期 1996-07-12
Measurement of serum carcino-associated antigen by anti-gallbladder mAb in digestive tract cancer
, 百拇医药
Gang Liang, Yuan-Ming Wang, Shu-You Peng, Zheng-Rong Shen and Fan Zhou
Department of Surgery, the Second Affiliated Hospital of Zhejiang Medical University, Hangzhou 310009, Zhejiang Province, China
Abstract
AIM To diagnose digestive tract cancer serologically using anti-gallbladder mAb.
METHODS Gallbladder associated antigen (GBAA) levels were measured by mAb for gallbladder cancer and conA in patients with digestive tract cancer, benign diseases and healthy controls. Serum CEA, AFP, CA19-9 and CA 242 levels were also measured simultaneously in the subjects.
, 百拇医药
RESULTS The GBAA positive rates were as follows: 2.2% in normal controls, 13.3% in benign diseases, 92.3% in gallbladder cancer, 20.9% in hepatocelullar cancer, 9.1% in pancreas cancer, 50% in gastric cancer, 41.6% in colon cancer, 50% in bile duct cancer, 100% in cancer of the ampulla of Vater. In gallbladder cancer, hepatocelullar cancer, pancreas cancer, gastric cancer, colon cancer, bile duct cancer, ampulla of Vater cancer, CEA positive rates were 7.6%, 3.8%, 33.3%, 21.5%, 33.3%, 0% and 0% respectively, AFP positive rates 0%, 57%, 6%, 0%, 0% and 0%, respectively, CA19-9 positive rates 62%, 3.8%, 77.7%, 0%, 50%, 0% and 50%, respectively, and CA242 positive rates 62%, 0%, 66.6%, 0%, 50%, 0% and 25% respectively. The sensitivity and specificity in diagnosis of gallbladder cancer by GBAA were 92.3% and 94.1% respectively. The sensitivity in diagnosis of tumors of non-gallbladder cancer was 32%. GBAA was also valuable for post-operative or recurrence monitoring. No correlation was found between the GBAA and tumor sizes, infiltration, differentiation and metastasis.
, 百拇医药
CONCLUSION Serum GBAA is useful for diagnosing gallbladder cancer, without cross reaction between GBAA and CEA or AFP.
Subject headings Gallbladder neoplasms Digestive system neoplasms Antibody, monoclonal CA_19_9 antigen/analysis Carcinoembryonic antigen/analysis α-fetoproteins/analysis
Liang G, Wang YM, Peng SY, Shen ZR, Zhou F. Measurement of serum carcino-associated antigen by anti-gallbladder mAb in digestive tract cancerXin Xiaohuabingxue Zazhi, 1997;5(5):323-324
, 百拇医药
摘 要目的 应用胆囊癌mAb对消化道肿瘤患者进行血清学诊断.
方法 采用胆囊癌mAb及刀豆蛋白A检测206例各种消化道肿瘤(胆囊癌13例、肝癌39例、胃癌39例、胰腺癌15例、胆管癌5例、壶腹部癌6例，结肠癌14例).良性疾病(30例)和健康供血者(45例)血清中胆囊癌相关抗原(GBAA)进行测定；同时测定CEA，AFP，CA19-9及CA242含量.
结果 正常人血清GBAA阳性率2.2%，良性疾病13.3%，胆囊癌GBAA 92.3%，CEA7.6%，AFP 0%，CA19-9 62%,CA242 62%;肝癌GBAA 20.9%，CEA3.8%，AFP 57.6%，CA19-9 3.8%,CA242 0%;胰腺癌GBAA 9.1%，CEA 33.3%，AFP 0%，CA19-9 77.7%,CA242 66.6%;胃癌GBAA 50%，CEA 21.5%，AFP 6%，CA19-9 0%,CA242 0%;大肠癌GBAA 41.6%，CEA 33.3%，AFP 0%，CA19-9 50%,CA242 50%;胆管癌GBAA50%，CEA 0%， AFP 0%，CA19-9 0%，CA242 0%；壶腹部癌GBAA 100%，CEA 0%，AFP 0%，CA19-9 50%,CA242 25%. GBAA对胆囊癌诊断敏感性92.3%，特异性94.1%，对非胆囊癌肿瘤诊断敏感性32%.GBAA对肿瘤术后及复发监测有一定价值.GBAA与胆囊癌肿瘤大小，浸润程度、组织分化及有无转移无相关性.
, http://www.100md.com
结论 GBAA单抗术前对胆囊癌诊断较高；GBAA单抗与CEA、AFP无交叉反应；临床上测定GBAA可作为CEA，AFP，CA19-9及CA242的补充，为临床诊断及普查提供了新的肿瘤标记物.
主题词 胆囊肿瘤 消化系统肿瘤 抗体，单克隆 CA-19-9抗原/分析 癌胚抗原/分析 甲胎蛋白/分析
梁刚, 王远明, 彭淑牖, 沈正荣, 周凡.胆囊癌mAb检测消化道肿瘤血清癌相关抗原.新消化病学杂志,1997;5(5):323-324
应用肿瘤单克隆抗体(monoclonal antibody, mAb)对恶性肿瘤患者进行血清学诊断是当今临床重点研究课题之一.中晚期胆囊癌生存率低，若早期诊断，可提高手术切除率，延长患者生命.作者自制胆囊癌mAb，建立ELISA法并对消化道肿瘤、良性疾病及健康供血者血清中胆囊癌相关抗原(GBAA)测定.同时对消化道肿瘤患者测定CEA，AFP，CA19-9，CA242，比 较其敏感性和特异性.
, http://www.100md.com
1 对象和方法
1.1 对象
抗体：以胆囊癌细胞作抗原，免疫Balb/c小鼠，与NS-1骨髓瘤细胞融合，获得分泌抗胆囊癌抗原(GBAA)的单克隆抗体.辣根过氧化酶(HRP.Sigma).CNBr-Sepharose 4B(Pharmacia).刀豆蛋白A(con A, Sigma).CEA试剂盒，AFP试剂盒(Syntron USA),CA19-9试剂盒(Sorin)，CA242试剂盒(Pharmacia). 血清标本：共206例，女性70例，男136例，平均年龄41岁. 其中胆囊癌13例，肝癌39例，胃癌39例，胰腺癌15例，胆管癌5例，壶腹部癌6例，结肠癌14例.良性疾病30例，其中慢性胆囊炎17例，急性胰腺炎8例，胃溃疡2例，肝脓肿2例，肝血管瘤1例，正常人45例.空腹抽血经离心分离血清，编号-20℃保存.
, 百拇医药
1.2 方法 GBAA测定：用150mg/L Con A包被微孔，4℃过夜，次日取出，洗涤3次，加入待检血清4℃反应6h，洗涤3次，加入纯化GBAA抗原标记的HRP，37℃ 1h，洗涤3次显 色，测各孔OD值.CEA，AFP，CA19-9，CA242均采用酶免疫法测定，按试剂说明书操作.统计学处理 采用SPSS统计软件包，卡方检验.Fisher's exact test.
2 结果GBAA OD值为正常人的x±2s判为阳性；正常值CEA≤2.5μg/L，AFP≤20μg/L， CA19-9≤37Ku/L，CA242≤17Ku/L.消化道肿瘤血清中肿瘤标志物表达阳性率见表1.
表1 消化道肿瘤血清中肿瘤标志物表达阳性率 (%)
| 肿瘤标记 | 胆囊癌 | 胃癌 | 肝癌 | 胰腺癌 | 大肠癌 | 胆管癌 | 壶腹部癌 |
| CEA | 7.6(1/13) | 21.5(3/11) | 3.8(1/26) | 33.3(3/9) | 33.3(2/6) | 0 | 0.0 |
| AFP | 0.0 | 60(1/16) | 57.6(15/26) | 0 | 0 | 0 | 0.0 |
| CA19-9 | 62.0(8/13) | 0.0 | 3.8(1/26) | 77.7(7/9) | 50.0(3/6) | 0 | 50.0(1/2) |
| CA242 | 62.0(8/13) | 0.0 | 0.0 | 66.6(6/9) | 50.0(3/6) | 0 | 25.0(1/4) |
| GBAA | 92.3(12/13) | 50.0(14/28) | 20.9(9/43) | 9.1(1/11) | 41.6(5/12) | 50(1/2) | 100(2/2) |
, 百拇医药
非肿瘤患者和人群血清中GBAA表达阳性率，正常人为1/45，良性疾病为4/30，假阳性6.7%，特异性93.3%；其中胆囊炎阳性率为1/11，与胆囊癌比较12/13，差别非常显著(P<0.01)，敏感性92.3%，特异性94.1%；非胆囊癌肿瘤的阳性率为32/100，差别也很显著(P<0.01). 研究发现术前GBAA阳性，术后切除的GBAA转阴31例；如不能切除则GBAA仍然阳性13例.如肿瘤复发则GBAA转阳性8例.GBAA与胆囊癌肿瘤大小、浸润程度、组织分化及淋巴转移无相关.
3 讨论胆囊癌居消化道肿瘤第6位，且近年有上升趋势.晚期胆囊癌预后极差，欲提高患者的预后，只有早期诊断，早期治疗胆囊癌.CEA，CA19-9及CA242术前诊断胆囊癌已有不少报道[1，2]，但缺乏敏感性和特异性.本研究发现GBAA在胆囊癌、胃癌、肝癌、胰腺癌、大肠癌、胆管癌及壶腹部癌阳性率分别为92.3%，50.0%，20.9%，9.1%、41.6%，50.0%及100%.GBAA对胆囊癌的敏感性92.3%，特异性94.1%，GBAA对临床诊断胆囊癌有较大价值. GBAA假阳性为6.7%.正常人群中出现少数假阳性，其原因有待进一步探讨.同时发现GBAA对胆管癌、壶腹癌肿瘤诊断有一定价值，由于例数少，有待于进一步研究.GBAA结合CA19-9或CA242有助于胆囊癌与胰腺癌之间的鉴别诊断.可见抗胆囊癌单克隆抗体可作为CEA，CA19-9的补充，为开展对胆囊癌的普查、临床诊断提供了一种新的肿瘤标志物.各种肿瘤 如术前GBAA阳性，手术切除后均转阴性，如肿瘤未切除，则GBAA持续阳性.发现术后复发(特别是胆囊癌患者)，则GBAA阳性.化疗后，GBAA测定为阴性，但观察例数不多，GBAA能否作为化疗后疗效判断需要进一步研究.肿瘤标志物应能指示出肿瘤大小、浸润程度、有无转移及病情的发展情况[3].本文研究GBAA与胆囊癌的肿瘤大小、组织类型、浸润程度及淋巴结转移关系无相关性.分析原因可能有：①病例数少，统计学处理无差异；②手术记录数字不精确；③肉眼无法判断有无微小转移灶.
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