Early detection of pancreatic cancer

The diagnosis of pancreatic cancer associates an appalling significance.Detection of preinvasive stage of pancreatic cancer will ameliorate the survival of this deadly disease.Premalignant lesions such as Intraductal Papillary Mucinous Neoplasms or Mucinous Cystic Neoplasms of the pancreas are detectable on imaging exams and this permits their management prior their invasive development.Pancreatic intraepithelial neoplasms(PanIN)are the most frequent precursors of pancreatic adenocarcinoma(PDAC),and its particular type PanIN high-grade represents the malignant non-invasive form of PDAC.Unfortunately,PanINs are not detectable on radiologic exams.Nevertheless,they can associate indirect imaging signs which would rise the diagnostic suspicion.When this suspicion is established,the patient will be enrolled in a follow-up strategy that includes performing of blood test and serial imaging test such as computed tomography or magnetic resonance imaging,which will cost in the best-case scenario a burden of healthcare systems,and potential mortality in the worst-case scenario when the patient underwent resection surgery,worthless when there is no moderate or high grade dysplasia in the final histopathology.This issue will be avoid having at its disposal a diagnostic technique capable of detecting high-grade PanIN lesions,such is the cytology of pancreatic juice obtained by nasopancreatic intubation.Herein,we review the possibility of detection of early malignant lesions before they become invasive PADC.

Detection of k-ras gene point mutation in fine needle aspiration and pancreatic juice by sequence special primer method and its clinical significance

INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected the presence of k-ras gene

Evaluation of diagnostic cytology via endoscopic naso-pancreatic drainage for pancreatic tumor

AIM: To evaluate the usefulness of cytology of the pancreatic juice obtained via the endoscopic naso-pancreatic drainage tube(ENPD-C).METHODS: ENPD was performed in cases where a diagnosis could not be made other than by using en-doscopic retrograde cholangiopancreatography and in cases of pancreatic neoplasms or cystic tumors, includ-ing intraductal papillary mucinous neoplasm(IPMN) suspected to have malignant potential. 35 patients(21 males and 14 females) underwent ENPD between January 2007 and June 2013. The pancreatic duct was imaged and the procedure continued in one of ENPD-C or ENPD-C plus brush cytology(ENPD-BC). We checked the cytology result and the final diagnosis.RESULTS: The mean patient age was 69 years(range, 48-86 years). ENPD-C was performed in 24 cases andENPD-C plus brush cytology(ENPD-BC) in 11 cases. The ENPD tube was inserted for an average of 3.5 d. The final diagnosis was confirmed on the basis of the resected specimen in 18 cases and of follow-up findings at least 6 mo after ENPD in the 18 inoperable cases. Malignancy was diagnosed in 21 cases and 14 patients were diagnosed as having a benign condition. The ratios of class Ⅴ/Ⅳ:Ⅲ:Ⅱ/Ⅰ?findings were 7:7:7 in malignant cases and 0:3:11 in benign cases. The sensitivity and specificity for all patients were 33.3% and 100%, re-spectively. The cytology-positive rate was 37.5%(6/16) for pancreatic cancer. For IPMN cases, the sensitivity and specificity were 33% and 100%, respectively.CONCLUSION: Sensitivity may be further increased by adding brush cytology. Although we can diagnosis cancer in cases of a positive result, the accuracy of ENPD-C remains unsatisfactory.

Preventing pancreatic fistula after distal pancreatectomy: An invagination method

Following an increase in the use of the GIA stapler for treating a pancreatic stump, more techniques to prevent postoperative pancreatic juice leakage have been required. We describe one successful case using our new technique of invaginating the cut end of the pancreas into the stomach to prevent a pancrea-tic fistula(PF) from occurring. A 50-year-old woman with pancreatic cancer in the tail of the pancreas underwent distal pancreatectomy, causing a grade A PF. We resected the distal pancreas without additional reinforcement to invaginate the stump into the gastric posterior wall with single layer anastomosis using a 3-0 absorbable suture. The drain tubes were removed on the third postoperative day. Although a grade A PF was noted, the patient was discharged on foot on the eleventh postoperative day. Our technique may be a suitable method for patients with a pancreatic body and tail tumor.

Detecting K-ras and p53 gene mutation from stool and pancreatic juice for diagnosis of early pancreatic cancer

OBJECTIVE: To explore new methods for the early diagnosis of pancreatic cancer through detection of K-ras and p53 mutations in pancreatic juice and stool. METHODS: 201 patients in PUMC Hospital from 1994 - 2000 and 60 control individuals were enrolled in this study. K-ras point mutation was detected by PCR-RFLP while p53 mutation was detected by PCR-SSCP. RESULTS: K-ras mutation was found in pancreatic juice in 87.8% (36/41) of pancreatic cancer patients and 23.5% (4/17) of benign pancreatic disease patients. In 261 stool specimens, amplification found mutations successfully in 235 patients (90%). K-ras mutation was found in stool in 88% (66/75) of pancreatic cancer patients, 51.1% (24/47) of benign pancreatic disease patients and 19.6% (9/46) of normal individuals. p53 mutation was found in pancreatic juice in 47.4% (18/38) of pancreatic cancer patients and 12.5% (2/16) of benign pancreatic disease patients. p53 mutation was found in stool in 37.1% (23/62) and 19.1% (4/21) of chronic pancreatitis patients. CONCLUSION: K-ras mutation in pancreatic juice has higher diagnosis sensitivity and specificity, and therefore may be used as a supplement in the diagnosis of pancreatic cancer. Detection of K-ras mutation combined with p53 mutation in stool can aid in the screening of pancreatic cancer.

胰液K-ras12密码子点突变检测对胰腺癌诊断的临床价值

目的 :探讨胰液中 K- ras 12密码子点突变对胰腺癌诊断的临床应用价值。 方法 :采用内镜下置鼻胰管方法收集 2 0例胰腺疾病的胰液标本 ,聚合酶链反应 -限制性片段长度多态性分析 (PCR- RFL P)检测胰液 K- ras基因第 12密码子点突变 ,与肿瘤标记物 CA 19- 9及 CEA检测结果比较。结果 :12例胰腺癌患者胰液标本中 K- ras突变率为 5 8.3% (7/ 12 ) ;8例慢性胰腺炎患者标本 K- ras突变率为 12 .5 % (1/ 8) ,胰腺癌胰液 K- ras基因突变检测的敏感性为 5 8.3% ,特异性为 87.5 % ,阳性预示值为 87.5 % ,阴性预示值为 5 8.3%。同一组病例以血清 CA19- 9>37U / m l、CEA >4 .5μg/ ml为阳性界值。胰腺癌血清CA19- 9、CEA阳性率分别为 5 8.3% (7/ 12 )、4 1.7% (5 / 12 )。结论 :K- ras基因突变与胰腺癌相关性好 ,胰液中 K- ras 12密码子突变率高 ,特异性强 ,为胰腺癌早期诊断的初步筛选提供了手段。

胰液中端粒酶及亚单位检测对胰腺癌的诊断价值

目的 探讨检测胰液中端粒酶逆转录酶 (hTERT)mRNA表达和端粒酶活性对胰腺癌的诊断与鉴别诊断价值。方法 对比分析 2 4例胰腺癌和 14例慢性胰腺炎患者胰液中的hTERTmRNA和端粒酶活性的检测结果。结果 hTERTmRNA在胰腺癌中表达阳性率为 87.5 % ( 2 1/ 2 4) ;慢性胰腺炎中阳性率为 2 1.4% ( 3 / 14 ) (P <0 .0 0 1)。胰腺癌与慢性胰腺炎胰液中端粒酶阳性检出率分别为83 .3 % ( 2 0 / 2 4) ,2 8.6% ( 4 / 14 ) (P <0 .0 0 1)。结论 胰液中检测hTERTmRNA表达和端粒酶活性对胰腺癌诊断与鉴别诊断有一定价值。

胰管刷检细胞端粒酶活性检测诊断胰腺癌的临床意义

目的 :探讨胰管刷检细胞端粒酶活性检测在胰腺癌诊断中的价值。 方法 :对 2 7例胰腺疾病患者采用 ERCP下胰管细胞刷获取标本 ,H- E染色 ,进行病理学观察 ;其中 2 1例采用 PCR- TRAP- EL ISA法测端粒酶活性 ,并与常规细胞学及血清CEA、CA 19- 9检查作比较。 结果 :胰腺癌患者端粒酶活性水平明显高于慢性胰腺炎患者 ,以端粒酶活性 D>0 .2为阳性界限值 ,胰腺癌端粒酶活性阳性率为 77.8% ,慢性胰腺炎无 1例端粒酶阳性 ,胰管刷检细胞端粒酶活性检测和细胞学诊断总符合率为 77.8% ,端粒酶活性与胰腺癌部位未见明显相关。结论 :胰管刷检标本细胞学检查的同时结合端粒酶活性检测是提高胰腺癌诊断水平的有效方法。

胰腺癌患者手术标本及胰液脱落细胞端粒酶活性的检测

目的 研究胰腺良、恶性患者手术标本 ,经ERCP所获胰液脱落细胞的端粒酶活性的表达 ,探索胰腺癌诊断的新方法。方法 采用TRAP ELISA的方法检测胰腺癌细胞株、胰腺良恶性疾病组织物、胰液细胞的端粒酶活性。结果 6株胰腺癌细胞株均检测到了端粒酶活性 ,胰腺癌手术标本端粒酶的阳性率为 63 % (5 /8) ,而 4例正常胰腺组织和 2例胰腺良性疾病患者的胰腺标本未检测到端粒酶活性。胰腺癌患者的胰液细胞中 ,有 5 8% (11/19)检测到有端粒酶活性的表达 ,而胰腺良性疾病也有高达 62 5 % (5 /8)的阳性率 ,两者无差别。结论 胰腺癌组织标本中有较高的端粒酶活性表达 ;胰液脱落细胞检测端粒酶活性无助于鉴别胰腺良恶性疾病。

胰腺癌胰液蛋白质组学分析

目的:利用比较蛋白质组学方法鉴定胰腺癌早期诊断的蛋白标志。方法:采用双向电泳(2-DE)分离比较9个胰腺导管腺癌患者和9个非胰腺癌患者的胰液蛋白质表达谱,用肽质量指纹图谱(PMF)为基础的基质辅助激光解析离子化飞行时间串联质谱(MALDF-TO-F-MS/MS)对差异蛋白进行鉴定。采用蛋白质印迹方法进一步验证蛋白质MMP-9和DJ-1在胰液中的表达。结果:2-DE显示共24个蛋白质点出现>2.0倍的表达差异,上调14个,下调10个。蛋白质印迹检测显示在胰腺癌胰液中高表达MMP-9和DJ-1蛋白。结论:MMP-9和DJ-1可能成为通过内镜下逆行胰胆管造影术(ERCP)筛查胰腺癌高危患者及早期诊断胰腺癌的潜在的胰液肿瘤标志。

检测胰液中K-ras基因突变对胰腺癌诊断的作用

目的:检测经逆行胰胆管造影(ERCP)过程中所获胰液中的K-ras基因突变。探讨其在胰腺癌诊断中的作用和意义。方法:收集ERCP过程中注射促胰泌素后的胰液,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)的方法检测胰腺良、恶性疾病胰液上清和细胞中的K-ras基因突变。结果:胰腺癌病例的胰液上清液中K-ras基因突变检出率为60.0%(18/30),细胞中的检出率为76.7%(23/30)。胰腺良性疾病胰液细胞中有15.4%(2/13)的K-ras基因突变,上清液和其他非胰腺疾病对照组胰液中无K-ras基因突变检出。胰腺癌患者的胰液中检测到K-ras基因突变明显高于胰腺良性疾病患者和对照组(P<0.001)。K-ras基因突变检出率与肿瘤的部位及大小无明显关系。结论:胰液K-ras基因突变检出对于胰腺癌的诊断具有一定作用。

胰液中p16基因启动子区5′CpG岛甲基化改变及其意义

目的:探讨胰液中p16基因启动子区5′CpG岛甲基化检测在胰腺癌诊断中的价值。方法:2005～2006年30例来自长海医院消化内镜中心的胰腺疾病患者入选,采用内镜逆行胆胰管造影(ERCP)下放置鼻胰管收集胰液.沉渣涂片H-E染色进行细胞学检查.甲基化特异性PCR(MSP)法检测胰液及组织中p16基因甲基化状态。结果:单纯胰液细胞学检查诊断胰腺癌的敏感性为40%(8/20).特异性为100%(10/10).阳性预测值为100%(8/8),阴性预测值为45.4%(10/22).诊断准确性60.0%(18/30)。所有胰液标本均成功抽提出DNA.30例胰腺疾病患者胰液标本进行p16基因启动子区5′CpG岛甲基化检测.其中20例胰腺癌患者胰液中p16基因甲基化率为35%(7/20).慢性胰腺炎和胰腺囊腺瘤患者胰液中无p16基因甲基化。胰液中p16基因甲基化与常规细胞学联合诊断胰腺癌的敏感性为55%(11/20),特异性为100%(10/10).阳性预测值为100%(11/11).阴性预测值为52.6%(10/19).诊断准确性70%(21/30)。结论:鼻胰管收集的胰液可用于分子生物学检测.检测胰液中p16基因甲基化改变与细胞学联合更有助于胰腺癌的诊断。

胰腺癌早期诊断研究的现状与展望

胰腺癌是一种严重危害人类健康的常见恶性肿瘤,其早期诊断和手术治疗可显著提高患者生存率,明显改善预后。近年来对胰腺癌的肿瘤标志物进行了大量研究,CA19-9、CA50、CA242、CA494等均是目前临床诊断胰腺癌的常用标志物,但迄今为止,仍未找到一种对早期胰腺癌有足够敏感性、特异性的肿瘤标志物来进行肿瘤筛查。一些细胞因子如转化生长因子(TGF)、巨噬细胞抑制因子-1(MIC-1)、可溶性细胞间黏附分子-1(sICAM-1)、可溶性血管内皮细胞黏附因-1(sVCAM-1)和P选择素等在胰腺癌的诊断、治疗监测及预后评估中具有重要作用。B超检查具有简便、经济、无创、无痛的特点,是胰腺癌的首选诊断方法。单独选用CT、螺旋CT、磁共振胰胆管成像(MRCP)、超声内镜(EUS)等在胰腺癌的早期诊断上各有其优势与不足。采用逆行胰胆管造影(ERCP)收集胰液、刷取脱落细胞行细胞学、K-ras基因和端粒酶检查是胰腺癌早期诊断的发展方向。单独某项影像学检查或血清指标的特异性和敏感性尚不能满足临床诊断早期胰腺癌的需要,因此,需要寻找新的肿瘤标志物,并建立影像学、脱落细胞、癌基因、抑癌基因、端粒酶等联合诊断的方法来进一步提高胰腺癌的诊疗水平。

胰液细胞学及肿瘤相关标志物联合检测在胰腺癌诊断中的价值

目的通过内镜抽取纯胰液检测细胞学、K-ras基因点突变及端粒酶活性,探索早期胰腺癌的诊断方法。方法58例胰腺癌高危病人,单纯十二指肠镜收集胰液后,分别行巴氏染色(Papanicolaou)作细胞学检查、半巢式PCR检测K-ras基因12密码子点突变及PCR-TRAP检测端粒酶活性。结果5例胰液中发现可疑恶性细胞(8.62%),K-ras基因点突变阳性22例(37.9%),端粒酶活性弱阳性8例(13.8%),三者均阳性4例(6.89%),K-ras基因点突变及端粒酶活性两者阳性7例(12.06%),1例细胞学阳性但K-ras基因点突变及端粒酶活性均阴性(1.72%)。结论内镜抽取纯胰液联合检测细胞学、K-ras基因点突变及端粒酶活性对临床诊断早期胰腺癌具有重要价值。

Stable expression of antisense hTR inhibits in vitro pancreatic cancer cell growth

OBJECTIVE: To clarify growth inhibition in pancreatic cancer cells by interference with the hTR component of the telomerase reverse transcriptase enzymatic complex. METHODS: A 593 bp full length hTR cDNA was subcloned into a mammalian expression vector pcDNA3.1(-) in the antisense orientation to construct an antisense hTR expression plasmid. These were introduced into panc1 cells, a human pancreatic carcinoma cell line, by lipofectin and G418-resistant stable transformants were expanded. Resulting stable clones were screened for the presence of the hTR insert by PCR with T7 and BGH reverse primers located on the flanks of the multiclonal site of the pcDNA3.1 vector. Cell growth rate, hTR expression, telomerase activity and anchorage-independent growth properties were analyzed. RESULTS: Significant downregulation of endogenous hTR was evident in the antisense-hTR transformed cells and telomerase activity was markedly decreased compared to control cells in standard TRAP assays. Furthermore, cell proliferation and the anchorage-independent growth ability in antisense-hTR expressing cells were significantly decreased compared with control parental cells. However, no crisis or senescence phenomena were observed. CONCLUSIONS: These data indicate that hTR may be a critical component of human telomerase activity and suggest that downregulation of the RNA component of human telomerase is a possible target for anticancer strategies.

胰腺癌患者胰液中端粒酶活性表达及临床意义

采用 DCR- ELISA和 PCR- SSCP两种方法检测胰腺癌患者胰液中端粒酶的活性。结果 31例胰腺癌患者中 ,19例端粒酶活性阳性 ;3例胰腺良性肿瘤患者中 ,1例端粒酶活性阳性 ;6例非胰腺疾病患者的端粒酶活性均为阴性。另外检测了 4例胰腺癌患者癌细胞株的端粒酶活性 ,结果均呈阳性。结果显示 ,胰腺癌患者胰液中端粒酶活性呈高表达 ,与胰腺良性肿瘤及非胰腺疾病相比差异有显著性 ( P<0 .0 5)。

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。

胰液端粒酶活性检测在胰腺疾病鉴别诊断中的价值

目的 :探讨端粒酶活性推测在胰腺癌早期诊断中的价值。方法 :4 0例胰腺癌和 31例慢性胰腺炎患者均行ERCP抽取胰液 ,采用TRAP PCR SSCP和TRAP PCR ELISA两种方法分别进行定性、定量分析 ,同时检测胰腺癌细胞株 4株 ,正常胰液和正常胰腺组织测定其端粒酶活性做为对照。结果 :端粒酶定性分析 :胰腺癌组胰液中阳性率 67 5 0 % ( 2 7/40 ) ;慢性胰腺炎组 4 1 93% ( 13/31) ;胰腺癌细胞株全部为阳性 ,阳性率 10 0 % ( 4 /4) ;正常胰腺和胰腺组织均呈阴性 ( 0 /5 )。端粒酶定量测定 :胰腺癌组为1 4 75± 0 4 67;慢性胰腺炎组为 0 35 8± 0 4 79。胰腺癌细胞株为 1 84 2± 0 2 5 6。胰腺癌组胰液中端粒酶阳性率明显高于慢性胰腺炎组 ,P <0 0 5 ;胰腺癌患者组 ,高于慢性胰腺炎组 ,P <0 0 1。结论 :胰液中端粒酶活性测定可作为胰腺良恶性疾病鉴别诊断的重要依据 。

胰腺良恶性病变组织中hTERT表达及临床病理意义

目的 研究胰腺良恶性病变组织中人端粒酶反转录酶hTERT表达意义。方法 石蜡包埋切片ABC免疫组化法。结果 慢性胰腺炎阳性率和评分明显低于癌旁上皮和胰腺癌 ,癌旁上皮也明显低于胰腺癌 ,阳性表达慢性胰腺炎或癌旁上皮呈不典型增生。高分化腺癌和未转移癌阳性率和评分均明显低于低分化腺癌和转移癌。结论 hTERT表达与胰腺癌发生及生物学行为存在密切关系 ,检测良性病变hTERT表达对预防和早期发现胰腺癌可能有临床应用价值。