胰液K-ras基因突变和血清CA19-9联合检测判断胰腺癌复发

目的研究胰液中K-ras12密码子点突变和血清CA19-9联合检测结果与胰腺癌病程的关系.方法测定32例临床及手术证实的胰腺癌患者血清CA19-9水平,同时采用内镜ERCP从胰管收集胰液标本,应用聚合酶链反应限制性片断长度多态性分析(PCR-RFLP)检测胰液K-ras基因12密码子点突变,分析K-ras12密码子点突变及血清CA19-9水平联合检测结果与胰腺癌临床特征和术后复发的关系.结果 (1)胰液中K-ras12密码子点突变率为56.3%,与肿瘤大小密切相关(P ＜ 0.05).K-ras12密码子点突变阳性、阴性表达病例3年复发率分别为66.7%和33.3%.(2)高血清CA19-9水平且K-ras12密码子点突变阳性组3年复发率为69.2%,而低血清CA19-9水平且K-ras12密码子点突变阴性组3年复发率为20.0%,两组差异显著(P ＜ 0.05).结论联合胰液中K-ras12密码子点突变和血清CA19-9检测可作为判断胰腺癌术后复发的有效指标,多因素分析对胰腺癌术后复发的判断更有价值.     

胰液K-ras基因突变和血清CA19-9联合检测判断胰腺癌复发

目的研究胰液中K-ras12密码子点突变和血清CA19-9联合检测结果与胰腺癌病程的关系。方法测定32例临床及手术证实的胰腺癌患者血清CA19-9水平,同时采用内镜ERCP从胰管收集胰液标本,应用聚合酶链反应限制性片断长度多态性分析(PCR-RFLP)检测胰液K-ras基因12密码子点突变,分析K-ras12密码子点突变及血清CA19-9水平联合检测结果与胰腺癌临床特征和术后复发的关系。结果 (1)胰液中K-ras12密码子点突变率为56．3％,与肿瘤大小密切相关(P<0．05)。K-ras12密码子点突变阳性、阴性表达病例3年复发率分别为66．7％和33．3％。(2)高血清CA19-9水平且K-ras12密码子点突变阳性组3年复发率为69．2％,而低血清CA19-9水平且K-ras12密码子点突变阴性组3年复发率为20．0％,两组差异显著(P<0．05)。结论联合胰液中K-ras12密码子点突变和血清CA19-9检测可作为判断胰腺癌术后复发的有效指标,多因素分析对胰腺癌术后复发的判断更有价值。     

内镜下胰液收集及肿瘤标记物检测对胰腺癌的诊断价值

目的研究内镜下置放鼻胰管收集胰液的方法及胰液细胞学检查、糖链抗原199(CA199)、癌胚抗原(CEA)水平和Kras基因12密码子点突变对胰腺癌诊断的临床应用价值。方法采用逆行胰胆管造影(ERCP)检查后置放鼻胰管收集20例胰腺癌、慢性胰腺炎患者胰液,采用聚合酶链反应限制性片段长度多态性分析(PCRRFLP)检测胰液Kras基因第12密码子点突变,放免法检测CA199和CEA。结果慢性胰腺炎和胰腺癌患者收集的胰液量分别为102ml±24ml(73ml～150ml)和86ml±27ml(56ml～140ml),胰液细胞学阳性率为8.3%。胰腺癌组胰液CA199值为(5620.58±1064.88)U/L,CEA值为(49.04±29.70)ng/L,明显高于慢性胰腺炎患者的CEA值(18.84ng/L±12.88ng/L),P<0.05。胰腺癌胰液Kras突变率为58.3%(7/12),慢性胰腺炎Kras突变率为12.5%(1/8),两者比较有显著性差异(P<0.05)。联合检测胰液Kras突变及CA199、CEA诊断胰腺癌的敏感性为83.3%,特异性为87.5%。结论联合检测胰液中肿瘤标记物对胰腺癌诊断和鉴别诊断具有较高的价值。     

胰液端粒酶和K-ras基因检测对胰腺癌的诊断价值

目的探讨胰液端粒酶活性与K-ras基因突变联合检测对胰腺癌患者诊断价值。方法收集25例胰腺癌和21例慢性胰腺炎患者胰液,K-ras基因突变采用聚合酶链反应-限制性酶切片段长度多态性(PCR-RFLP)检测,端粒酶活性检测采用端粒末端重复序列扩增技术(TRAP)。结果胰腺癌中K-ras基因突变率为88.0%(22/25),慢性胰腺炎中K-ras基因突变率仅为19.0%(4/21),P<0.01;在25例胰腺癌中端粒酶阳性检出率为84.0%(21/25);而21例慢性胰腺炎胰液中端粒酶阳性检出率为28.6%(6/21),P<0.01。K-ras基因突变或端粒酶活性阳性检测诊断胰腺癌的特异、敏感性分别为92%,66.7%;两者联合检测诊断胰腺癌的敏感性为80.0%,特异性达到85.7%。结论联合检测胰液中K-ras基因、端粒酶活性可提高胰腺癌诊断的敏感性和特异性,对胰腺癌筛查、诊断和鉴别诊断有一定临床意义。     

K-ras基因点突变对胰腺癌的诊断价值

背景传统的放射或超声检查早期诊断胰腺癌非常困难.目的研究胰腺良、恶性病变中的K-ras基因12密码子点突变情况,探讨K-ras基因点突变对早期胰腺癌的诊断价值.方法取23例胰腺癌和12例胰腺良性病变标本,经DNA提取后,行半巢式聚合酶链反应(PCR),扩增产物借助聚丙烯酰胺凝胶电泳检测有无K-ras基因点突变.结果胰腺癌和胰腺良性病变石蜡包埋组织的K-ras基因12密码子点突变率分别为65.2%(15/23)和33.3%(4/12)(P＜0.05).结论K-ras基因12密码子点突变的检测可能有助于胰腺癌,尤其是早期胰腺癌的诊断.     

十二指肠液中CA19—9水平与胰胆管癌关系的研究

目的探讨十二指肠肠液中CA19—9与胰胆管癌病程的关系。方法采用十二指肠肠镜从十二指肠中收集的胆胰液标本，测定48例临床及手术证实的胰胆管癌患者胰液CA19—9水平，研究胆胰液CA19—9水平与胰腺癌术后复发的关系。结果胆胰液CA19—9水平高组3年复发率为62．5％，而胆胰液CA19—9水平低组3年复发率为30．0％，两者差异有显著性（P〈0．05）。结论胆胰液CA19—9检测可作为判断胰腺癌术后复发的有效指标，对胰腺癌术后复发的判断更有价值。     

胰腺癌组织K-ras基因第12密码子突变的定量检测

目的 检测胰腺癌及相应癌旁组织K-ras基因第12密码子的突变量,分析其与胰腺癌临床病理参数的关系.方法 应用肽核酸(peptide nucleic acid,PNA)钳制双荧光标记探针的实时定量PCR法检测93例胰腺癌组织及其癌旁组织中K-ras基因第12密码子的突变拷贝数,以突变百分率表示突变量.K-ras基因突变百分率=K-ras突变型拷贝数/(K-ras野生型拷贝数+K-ras突变型拷贝数)×100%.结果 胰腺癌及其癌旁组织的K-ras基因第12密码子突变率分别为83.9%和65.6%,相差显著(P＜0.05);它们的突变量分别为(13.385±1.745)%和(2.246±0.728)%,相差亦显著(P＜0.05).K-ras基因第12密码子突变量与临床病理参数无关.结论 胰腺癌及相应癌旁组织不仅存在K-ras基因突变率的差异,亦存在K-ras基因突变量的差异.     

血清肿瘤标志物在结肠癌诊断中的应用价值

目的探讨血清肿瘤标志物癌胚抗原(CEA)、糖蛋白抗原(CA)50、CA199和CA724在结肠癌诊断中的临床价值。方法选取60例结肠癌患者为病例组,30例同期体检的健康人群为对照组,分析血清CEA、CA50、CA199和CA724的水平。结果结肠癌患者血清CEA、CA50、CA199和CA724的水平均明显高于对照组(P<0.05),且晚期结肠癌患者血清中此4种肿瘤标志物的水平显著高于早期结肠癌患者(P<0.05)。术后患者血清CEA、CA50、CA199和CA724的水平明显较术前降低(P<0.05),而术后发生复发转移时,其水平又明显升高,与术前无显著差异(P>0.05);将CEA、CA50、CA199和CA724联合检测用于诊断和复发转移检测,其阳性检出率明显高于单项指标检测(P<0.05)。结论 CEA、CA50、CA199和CA724在结肠癌的阳性检出、分期和复发转移中具有检测指示意义,且联合检测进行诊断更具优势。     

PCR-MASA检测胰腺癌外周血K-ras基因点突变的研究

目的了解胰腺癌外周血中K-ras基因点突变检测的临床价值.方法采用PCR-MASA法检测胰腺癌患者外周血中K-ras基因点突变.结果胰腺癌外周血标本中K-ras基因点突变率为38.1%(8/21),而所有被检测的急、慢性胰腺炎、胰岛素瘤、壶腹癌、十二指肠乳头癌、胆管癌及胆石症患者外周血标本均无K-ras基因突变.结论(1)PCR-MASA方法简捷、特异、敏感,扩增产物只需常规电泳、染色即可观察结果,无需酶切、杂交、放射性和非放射性显影;(2)对外周血标本检测K-ras基因第12位密码子有无突变,具有临床实用性,有助于判断胰腺病变良恶性及胰腺癌的早期诊断.     

外周血K-ras基因突变检测在胰腺癌诊断中的应用

目的 检测胰腺癌患者外周血K-rag基因第12、13密码子突变,探讨其对胰腺癌的诊断价值.方法 选取经病理证实的胰腺癌患者54例以及健康志愿者33例.抽取全部实验者外周血标本,抽提循环中的DNA,应用肽核酸钳制实时定量PCR法检测K-rag基因突变,并分析其与患者临床病理参数的相关性.结果 54名胰腺癌患者中,40例(74.1%)外周血标本可检测出K-ras基因第12或13密码子突变.33名健康志愿者外周血标本无一例检出K-ras基因突变.外周血中K-ras基因的突变与患者年龄、淋巴血管侵犯、肿瘤分化程度、肿瘤临床分期、CA19-9水平有关(P＜0.05),而与患者性别、肿瘤部位、肿瘤大小及病理类型无关.结论 肽核酸钳制实时定量PCR法检测外周血K-ras基因突变的敏感性高.外周血K-ras基因突变的检出提示肿瘤具有高侵袭性,可能预后不良.     

顺序特异性引物法快速检测胰腺癌K—ras基因点突变

目的：研究简捷、特异、敏感的检测胰腺癌K－ras基因点突变的方法及其在胰腺疾病中定性诊断的价值。方法：采用针对K－ras基因点突变方式（CGT、GAT、GTT）设计的顺序特异性引物（SSP），先后对胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液进行多聚酶链反应（PCR），扩增产物借助常规电泳和染色检测有无K－ras基因突变及突变方式。结果：胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液中K－ras基因点突变率分别为74．2％、95．1％、91．4％及94．1％，而所有被检测的慢性胰腺炎、胰岛素瘤、壶腹癌、胆管癌、十二指肠乳头癌及外伤胰腺的组织标本和胰液标本均无K－ras基因突变发生。结论：该检测法简便、快速、特异、敏感，具有临床实用性，可以作为鉴别胰腺肿块良恶性和诊断胰腺癌的一种方法。     

胰腺癌患者胰液中K-ras基因突变的检测及其意义

目的 通过对经逆行胰胆管造影（ＥＲＣＰ）所获胰液的Ｋ－ｒａｓ基因突变检测,以探索胰腺癌诊断的新方法。方法 应用聚合酶链反应（ＰＣＲ）－限制性片段长度多态性的方法检测胰腺良、恶性疾病组织物、胰液上清和细胞的Ｋ－ｒａｓ突变。结果 胰腺癌和胰腺良性疾病标本分别有７１％（１５／２１）,而胰腺良性疾病无一例有Ｋ－ｒａｓ的突变。用胰液细胞有４％ＰＣＲ未能获得成功,胰腺癌胰液细胞Ｋ－ｒａｓ的突变率为６５％（１１     

肿瘤标志物、K-ras和p53突变对胰腺癌诊断的价值

目的评价血清肿瘤标志物CA19-9、CA242、CA50、癌胚抗原和粪便K-ras以及p53基因突变对胰腺癌诊断的价值。方法收集2002年2月至2004年3月在北京协和医院、中国医学科学院肿瘤医院和沈阳军区总医院确诊的新发胰腺癌患者136例,良性消化系统疾病患者240例,进行血清肿瘤标志物和粪便K-ras、p53基因突变的检测。根据结果绘制不同检测方法的受试者工作特征(ROC)曲线,计算ROC曲线下面积,并确定最佳阳性分界值。结果血清CA19-9和CA242的ROC曲线下面积分别为0·855±0·031(95%可信区间0·794~0·916)和0·859±0·031(95%可信区间0·799~0·920),最佳阳性分界值分别为68U/ml和25U/ml,其诊断胰腺癌的敏感性分别为84·4%(98/116)和88·4%(84/95),特异性分别为84·3%(145/172)和79·1%(144/182)。粪便K-ras和p53基因突变诊断胰腺癌的敏感性分别为77·8%和27·8%,特异性分别为82·2%和95·2%。将粪便K-ras和p53基因突变与血清CA19-9和CA242测定相结合计算胰腺癌诊断评分,绘制有序分类资料的ROC曲线,其曲线下面积为0·946±0·017(95%可信区间0·912~0·980),最佳阳性分界值为2分。结论血清CA19-9及CA242对胰腺癌诊断具有相似价值;联合粪便K-ras及p53突变的检测,通过胰腺癌可能性积分,可以显著提高胰腺癌的诊断效率。     

Analysis of K-ras codon 12 point mutations using duodenal lavage fluid for diagnosis of pancreatic carcinoma

We evaluated the diagnostic significance of the K-ras point mutations at codon 12 in duodenal lavage fluid (DLF) compared with pure pancreatic juice (PPJ). The DLF was easily and safely collected by injecting distilled water into the duodenum and then aspirating through the working channel of the endoscope during endoscopic retrograde cholangiopancreatography. Two types of DLF are collected this way: DLF 1 is collected just after insertion of the endoscope into the duodenum and DLF 2 is collected after cholangiopancreatography and/or collection of the PPJ using secretin. Analysis of K-ras mutations was performed using enriched polymerase chain reaction. In patients with pancreatic carcinoma (PC), K-ras mutations were detected in 14 of 23 (60.9%) in DLF 1, 16 of 21 (76.2%) in DLF 2, 14 of 20 (70.0%) in PPJ, and 19 of 21 (90.5%) in either DLF 1 or DLF 2. In patients with noncancerous pancreatic diseases consisting of pancreatic cystic diseases and chronic pancreatitis, the incidence of K-ras mutations was 2 of 21 (9.5%) in DLF 1 and 7 of 19 (36.8%) in DLF 2. These values were lower than that in PPJ, and there was significant difference between the incidence in DLF 1 and PPJ. These results suggested that DLF may provide a new and useful material for analysis of K-ras codon 12 point mutations in the diagnosis of PC.     

Significance of K-ras codon 12 point mutation in pancreatic juice in the diagnosis of carcinoma of the pancreas

The significance of K-ras codon 12 point mutation in pancreatic juice in the diagnosis of carcinoma of the pancreas is still unclear. The aim of this study was to evaluate the significance of K-ras codon 12 point mutation in pancreatic juice in the diagnosis of carcinoma of the pancreas. All of the 78 reports written from 1988 to 1996 on K-ras point mutation of carcinoma, mucin-producing tumors, and hyperplastic epithelia of the pancreas in both surgical or autopsy specimens and pancreatic juice are reviewed. As results, in surgical or autopsy specimens, K-ras mutation was found in 81% of ordinary duct cell carcinoma and in 53% of mucin-producing tumor of the pancreas; this mutation was also found in hyperplastic epithelia in chronic pancreatitis (7%) and in autopsy cases without pancreatic diseases. In pancreatic juice, K-ras mutation was found in 72% of ordinary pancreatic carcinoma and in 53% of mucin-producing tumor, respectively. In conclusion, most previous reports have indicated that K-ras mutation in pancreatic juice is useful for a diagnosis of pancreatic carcinoma. However, since K-ras gene mutation was also detected in non-tumorous lesions, the diagnosis of pancreatic carcinomas is not necessarily correct if it is based solely on the detection of K-ras mutation in pancreatic juice. Future studies should focus on analyzing the amino acid sequence of K-ras mutation or the combination of this mutation with other parameters such as tumor markers in pancreatic juice, to enhance its specificity and accuracy.     

Diagnosis of pancreatic cancer by cytology and measurement of oncogene and tumor markers in pure pancreatic juice aspirated by endoscopy

BACKGROUND/AIMS: Cytological examination of pancreatic juice is useful in the diagnosis of an occult cancer of the pancreas. The early diagnosis of pancreatic carcinoma using traditional radiographic or ultrasonographic methods is extremely difficult. METHODOLOGY: In order to detect an early pancreatic cancer, cytological examination, measurement of tumor marker, and detection of K-ras point mutation were performed using the samples of pure pancreatic juice aspirated endoscopically in patients who had symptoms or findings that suggested pancreatic disease. RESULTS: By routine ERP-cytology, positive cytologic results were obtained in 15 (4%) out of 359 patients without a mass. With the aid of intra-operative cytodiagnosis, all 15 occult neoplasms of the pancreas were successfully resected. One patient died from another disease without evidence of recurrence. However, the other patients were alive with no evidence of recurrence for an average of 5.5 years following surgery. The patients who had negative ERP-cytology results were observed, but no further cases of pancreatic cancer were found. The CEA levels in the pure pancreatic juice were significantly higher in patients with pancreatic cancer than in those with pancreatitis. K-ras point mutation at codon 12 was detected not only in cases of pancreatic cancer, but also in cases of chronic pancreatitis as well as control subjects. CONCLUSIONS: Cytological examination of pancreatic juice is useful in the diagnosis of an early and potentially curable in situ cancer of the pancreas. The CEA levels in the pure pancreatic juice provided useful information for differentiating the pancreatic cancer from chronic pancreatitis. K-ras point mutation at codon 12 in pancreatic juice was considered to be useful in identifying patients at high risk for the development of pancreatic cancer.     

胰腺癌患者血浆k-ras基因与肿瘤标志物联合检测及其意义

目的 :了解胰腺癌患者血浆中肿瘤标志物水平和k ras基因突变情况 ,评价基因突变与肿瘤标志物联合检测对胰腺癌患者的诊断价值。方法 :收集经手术或病理确诊为胰腺恶性疾病患者 2 1例 ,ELISA检测血浆CA19 9、CA2 42、CA5 0、CEA水平 ,PCR RFLP检测k ras基因突变 ,并与 11例胰腺良性疾病患者对照。结果 :胰腺癌患者血浆中k ras基因突变率 73.7%,胰腺良性疾病k ras基因无突变。k ras基因突变检测的敏感性与特异性分别为 6 1.9%和10 0 %,血浆k ras、CA19 9、CA2 42联合检测的敏感性和特异性分别为 85 .7%和 71.9%。结论 :联合检测血浆中k ras基因与肿瘤标志物可提高胰腺癌诊断的敏感性 ,对胰腺癌筛查、诊断与鉴别诊断有一定的临床意义。     

PCR—MASA检测胰腺癌外周血K—ras基因点突变的研究

目的 了解胰腺癌外周血中K—ras基因点突变检测的临床价值。方法 采用PCR—MASA法检测胰腺癌患外周血中K—ras基因点突变。结果胰腺癌外周血标本中K—ras基因点突变率为38．1％(8／21)，而所有被检测的急、慢性胰腺炎、胰岛素瘤、壶腹癌、十二指肠乳头癌、胆管癌及胆石症患外周血标本均无K—ras基因突变。结论 (1)PCR—MASA方法简捷、特异、敏感，扩增产物只需常规电泳、染色即可观察结果，无需酶切、杂交、放射性和非放射性显影；(2)对外周血标本检测K—ras基因第12位密码子有无突变，具有临床实用性，有助于判断胰腺病变良恶性及胰腺癌的早期诊断。