胰腺癌与mdrl基因的相关性研究(英文)

目的通过研究未化疗的原发性胰腺癌 mdrl 基因 mRNA 及其蛋白产物 P-gp 的表达与肿瘤生物学特性的关系,以便指导胰腺癌的临床治疗。方法应用免疫组织化学、原位 PCR 技术对150例胰腺石蜡包埋标本(包括97例原发胰腺癌,32例胰腺炎及21例正常胰腺组织)组织中的 mdrl mRNA 和 P-gp 进行检测。结果 mdrl mRNA 和 P-gp 染色主要分布于正常胰腺小导管及肿瘤的导管上皮细胞的细胞膜和/或细胞浆内。胰腺癌和胰腺炎及正常胰腺组织中 mdrl mRNA 和 P-gp 表达阳性率之间存在着显著性差异(P<0.05),mdrl mRNA 和 P-gp 的阳性表达率与肿瘤的分化程度、浸润性和 TNM 分期等生物学特性有关(P<0.05),而与病人的年龄、性别、肿瘤的大小及部位等临床病理资料无关(P>0.05)。结论研究结果表明胰腺癌 mdrl 基因的表达与其“天然”耐药有关,检测胰腺癌 mdrl 基因表达对预测胰腺癌的化疗敏感性具有重要指导意义。同时它可能作为判断肿瘤的恶性程度及预后的一个有效生物学指标。     

LivinmRNA在胰腺癌中的表达及与Bcl-2相关性的研究

目的:探讨凋亡抑制基因Livin mRNA在胰腺癌组织中的表达及其与Bcl-2蛋白表达的关系.方法:采用逆转录-多聚酶链反应(RT-PCR)法检测52例胰腺癌、10例慢性胰腺炎和12例正常胰腺组织中Livin mRNA的表达;用免疫组织化学法检测胰腺癌组织中Bcl-2蛋白的表达.结果:Livin mRNA在胰腺癌中的表达率为71.2%,明显高于慢性胰腺炎和正常胰腺组织(P<0.01).Livin基因表达与胰腺癌的分化程度和淋巴结转移有关(P<0.05).Livin基因表达与Bcl-2蛋白表达显著相关.结论:Livin基因在胰腺癌的发生发展中起重要作用;Livin与Bcl-2的异常表达可能在胰腺癌的发生中起协同作用.     

肿瘤转移抑制基因KiSS-1在胰腺癌组织中的表达及意义

目的:检测KiSS-1mRNA及KiSS-1蛋白metastin在胰腺癌组织中的表达,探讨其在胰腺癌浸润及转移中的作用.方法:采用逆转录多聚酶链反应(RT-PCR)法及免疫组织化学S-P法检测42例胰腺癌,10例慢性胰腺炎及12例正常胰腺组织中KiSS-1mRNA及KiSS-1蛋白metastin的表达情况及与各种临床病理参数的关系.结果:KiSS-1mRNA及Kiss-1蛋白metastin在胰腺癌组中的阳性表达显著低于慢性胰腺炎组及正常胰腺组(P＜0.01);KISS-1mRNA和KiSS-1蛋白metastin的表达均与胰腺癌临床分期及淋巴结转移有关(P＜0.05);二者在胰腺癌组织中的表达存在明显相关性.结论:肿瘤转移抑制基因KiSS-1在抑制胰腺癌的浸润和转移过程中可能起着重要的作用.     

胰腺癌组织转录因子GATA-6表达及其意义的探讨

目的:探讨胰腺癌组织GATA-6的表达及其对胰腺癌发生、发展的可能作用及临床意义。方法:采用免疫组化检测33例胰腺癌、15例慢性胰腺炎和6例正常胰腺组织GATA-6蛋白的表达;免疫细胞荧光法检测胰腺癌细胞株Panc-1、BxPC-3和SW1990及正常胰腺导管上皮细胞H6C7的GATA-6蛋白表达情况;Real-time RT-PCR技术检测上述细胞株的GATA-6 mRNA表达;并分析GATA-6与胰腺癌临床病理参数的关系。结果:GATA-6蛋白在胞核中表达,胰腺癌组织GATA-6的阳性表达率(88.9%)显著高于慢性胰腺炎(40.0%)和正常胰腺组织(16.7%),P<0.01,但与性别、年龄、分化程度和临床分期均无显著性相关;3种胰腺癌细胞株均有GATA-6蛋白表达,而正常胰腺导管上皮细胞无表达;3种胰腺癌细胞株GATA-6 mRNA表达也明显高于正常胰腺导管上皮细胞,P<0.05。结论:GATA-6在胰腺癌组织及细胞中呈高表达,提示GATA-6在胰腺癌的发生、发展中可能起一定作用。     

胰腺癌组织存活素和极光B表达的相关性

目的探讨凋亡抑制蛋白存活素（survivin）和极光B(AURORA_B)在胰腺癌组织中的表达与生物学行为之间的关系及两者的相关性。方法用免疫组织化学SP法检测45例胰腺癌和8例慢性胰腺炎及7例正常胰腺组织切片的survivin和AURORA_B的表达。结果45例胰腺癌中survivin和AURORA_B蛋白的阳性表达率分别为75.55%和53.33%;两者在7例正常胰腺组织中均未发现阳性表达;两者在8例慢性胰腺炎组织中的阳性表达率分别为25%和0。survivin的表达与组织学分级有关（P<0.05）,而与临床分期和淋巴结转移关系不大（P>0.05）,AURORA_B的表达与临床分期和淋巴结转移有关（P<0.05）,而与组织学分级关系不大（P>0.05）;survivin与AURORA_B的表达密切相关（P<0.05）。两者的表达与患者的性别、年龄、肿瘤的大小及部位均无关。结论survivin和AURORA_B密切相关,可能在胰腺癌的发生、发展过程中起关键作用,可能为胰腺癌的治疗和预防提供了新的靶点。     

NDRG1与MMP-7在胰腺导管腺癌中的表达及意义

目的:检测胰腺导管腺癌中NDRG1及MMP-7的表达,探讨其与临床病理特征的关系,并分析二者在胰腺癌诊断及治疗中的意义。方法:胰腺导管腺癌标本86例,胰腺导管腺癌癌旁正常胰腺组织86例,应用免疫组化法检测NDRG1和MMP-7蛋白的表达情况,检测其表达水平与临床病理特征的关系及二者的统计学相关性。结果:NDRG1和MMP-7表达于胰腺导管腺癌导管内皮细胞膜及细胞质,平均光密度值分别为0.1934±0.0327及0.1876±0.0249,在癌旁正常胰腺组织中NDRG1、MMP-7平均光密度值分别为0.1323±0.0243及0.1156±0.0297,二者差异有统计学意义（P<0.01;P<0.05）。NDRG1高表达与肿瘤直径、肿瘤TNM分期、淋巴结转移呈正相关性（P<0.05;P<0.01;P<0.05）。MMP-7高表达与肿瘤直径、肿瘤TNM分期、淋巴结转移呈正相关性（P均<0.05）。Spearman rank相关分析结果显示,NDRG1及MMP-7呈统计学正相关性（r＝0.267,P<0.05）。结论:NDRG1和MMP-7蛋白表达增高与胰腺导管腺癌的发生、发展及侵袭转移有关。NDRG1和MMP-7在胰腺导管腺癌组织中的表达相关,二者联合检测可作为胰腺癌预后判断及治疗的靶向候选基因。     

乙酰肝素酶和MMP-9 在胰腺癌中的表达及临床意义*

目的:检测乙酰肝素酶（heparanase,HPA ）和基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）在胰腺癌、慢性胰腺炎、正常胰腺组织中的表达及相互关系,分析其与浸润转移的关系。方法:应用免疫组化SP法检测92例胰腺癌、24例慢性胰腺炎和10例正常胰腺组织中HPA 、MMP-9 的表达水平,分析两者的表达与临床病理指标的关系,建立Spearman 等级相关分析。结果:HPA 在胰腺癌中表达明显高于慢性胰腺炎和正常胰腺组织（P<0.05）,HPA 阳性表达与周围组织浸润、淋巴结转移呈正相关（P<0.05）,与性别、年龄及组织学分级无统计学相关（P>0.05）;胰腺癌组织中MMP-9 表达明显高于慢性胰腺炎和正常胰腺组织（P<0.05）,且MMP-9 阳性表达与周围组织浸润、远处转移正相关（P<0.05）,与性别、年龄、组织学分级无统计学相关（P>0.05）。HPA 与MMP-9 有较高的共同阳性表达率。HPA 和MMP-9 表达呈正相关（P<0.05）,其共同表达率与肿瘤对周围脏器浸润和淋巴结转移有相关（P<0.05）。 多因素分析显示HPA 和MMP-9 均非影响预后的独立因素。结论:HPA 、MMP-9 在胰腺癌组织中的表达可能与胰腺癌浸润、转移相关,可作为新的胰腺癌标记物用于联合检测,具有重要的临床意义。且MMP-9 可能上调HPA 表达水平,促进HPA 介导的肿瘤浸润转移。      

核因子κB在胰腺癌中的表达及与VEGF和p53表达的相关性

目的探讨核因子κB(NF-κB,p65)在胰腺癌组织中的表达及其与VEGF、p53蛋白表达的相互关系。方法电泳迁移率变动分析(EMSA)测定45例胰腺癌及癌旁组织、8例慢性胰腺炎和9例正常胰腺组织中NF-κB的活性,Western印迹法检测NF-κB、IκB-a蛋白表达;免疫组织化学法检测胰腺癌中VEGF、p53、p65蛋白表达。结果NF-κB在肿瘤组织中被激活;NF-κB在胰腺癌、旁组织、慢性胰腺炎、正常胰腺组织中阳性表达率分别为71.0%、20.0%、12.5%、0(P<0.05);NF-κB基因表达与胰腺癌的分化程度、临床分期、肿瘤大小无关(P>0.05),与淋巴结转移和周围浸润有相关性(P<0.05);转移癌中NF-κB蛋白表达与VEGF、p53蛋白表达显著相关。结论核因子κB基因在胰腺癌中过度表达并与VEGF、p53一起参与胰腺癌的浸润、转移过程。     

胰腺导管癌神经生长因子mRNA表达及意义

目的研究胰腺导管癌和慢性胰腺炎组织中神经生长因子(NGF mRNA)的表达及临床病理意义.方法 51例胰腺癌和10例慢性胰腺炎手术切除标本经40 g/L中性甲醛固定后常规制作石蜡包埋切片,NGF mRNA染色为原位杂交法.结果胰腺癌组织NGF mRNA阳性率(54.9%)明显高于慢性胰腺炎(10.0%),差异有高度显著性(χ2=6.76,P＜0.01);高分化腺癌、未转移胰腺癌NGF mRNA表达阳性率明显低于低分化腺癌(25.0%对84.2%;χ2=13.74,P＜0.01)和转移病例(31.3%对65.7%;χ2=5.27,P＜0.05), NGF mRNA与胰腺癌其他临床病理特征无明显相关.结论 NGF mRNA表达可能与胰腺癌发生、进展、转移及预后有较密切的关系,可能是胰腺癌发生、发展及预后的重要生物学标记物.     

胰腺癌组织中表皮生长因子受体、基质金属蛋白酶9的表达及其意义

目的 探讨胰腺癌组织中表皮生长因子受体(EGFR)、基质金属蛋白酶9(MMP-9)的表达及其 与胰腺癌临床病理特征的关系。方法 应用免疫组织化学SP法检测44例胰腺癌、相应癌旁胰腺组织、 13例慢性胰腺炎和7例正常胰腺组织中EGFR、MMP-9的表达,并分析其与临床病理特征的相关性。 结果 正常胰腺组织中均无EGFR及MMP-9的表达。慢性胰腺炎组织中EGFR、MMP-9的阳性表达 率分别为23.1%(3/13)和15.4%(2/13)。胰腺癌组织中EGFR、MMP-9的阳性表达率分别为61.4%(27/44) 和54.5%(24/44);相应癌旁组织中的阳性表达率分别为34.1%(15/44)和29.5%(13/44)。胰腺癌组织中 的EGFR及MMP-9阳性表达率均显著高于相应癌旁组织(P<0.05);胰腺癌组织及癌旁组织中EGFR及 MMP-9的表达均显著高于慢性胰腺炎组织和正常胰腺组织(P均<0.05)。EGFR和MMP-9的表达与胰腺 癌临床分期、分化程度、血管侵犯及淋巴结转移相关。结论 胰腺癌组织中EGFR的表达与MMP-9表 达密切相关,EGFR的表达与MMP-9的表达水平可作为了解胰腺癌生物学行为和判断预后的指标。     

Serum dickkopf-1 is a novel serological biomarker for the diagnosis and prognosis of pancreatic cancer

Purpose

To identify whether Dickkopf-1 (DKK1) could be a potential biomarker for early detection and prognosis in patients with pancreatic cancer (PC).

Methods

Serum was collected from 140 patients with pancreatic adenocarcinoma and 92 control patients without pancreatic adenocarcinoma. Serological levels of DKK1 were examined by enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity was compared with carbohydrate antigen 19-9 (CA19-9). A 2-year follow-up was monitored to evaluate the correlation between DKK1 serum levels and overall survival. The expression of DKK1 in PC tumor tissues was also evaluated using immunohistochemistry staining.

Results

Serum levels of DKK1 and CA19-9 were elevated in PC patients in the early-stage cases. These levels increased with the advancement of clinical stage. There was significant difference in DKK1 serum levels between early and advanced PC stages. Receiver operating characteristic curve (ROCC) analysis showed that DKK1 was significantly better than CA19-9 in differentiating patients with PC from the controls (area under the curve (AUC) 0.919 versus 0.853, respectively), especially in distinguishing early-stage cancer from chronic pancreatitis (CP). The expression of DKK1 in PC tissues correlated with its expression in serum samples. The overall survival rate was 24.4% in the group with higher DKK1 levels and was found to be significantly different from the group with lower DKK1 levels (33.3%).

Conclusion

DKK1 may be a novel diagnostic/prognostic biomarker for PC.     

载体表达小干扰RNA逆转卵巢癌的多药耐药

目的:探讨载体表达的小干扰RNA(smallinterferingRNA,siRNA)逆转卵巢癌细胞多药耐药的可行性。方法:浓度梯度诱导法建立人卵巢癌阿霉素耐药细胞株OVCAR/AR;脂质体介导将MDR1特异性siRNA的表达载体(pSN/mdr1a和pSN/mdr1b)转染OVCAR/AR细胞;实时定量RT-PCR检测MDRlmRNA的表达;流式细胞术检测P-gp的表达,罗丹明试验检测P-gp的药物转运功能;MTT法检测OVCAR/AR细胞对化疗药的抵抗性。结果:转染pSN/mdr1a和pSN/mdr1b后,OVCAR/AR细胞的MDR1mRNA和P-gp的表达均显著下降,P-gp的转运功能减少,OVCAR/AR细胞对阿霉素、泰素的耐药性逆转。结论:载体表达的siRNA可持久有效地抑制卵巢癌耐药细胞MDR1mRNA和P-gp的表达,并逆转其多药耐药。     

多药耐药基因mdr-1在肝门部胆管癌组织中的表达及其意义

目的：检测肝门部胆管癌新鲜组织中多药耐药基因（ｍｄｒ－１）ｍＲＮＡ表达，探讨其与肝门部胆管癌临床病理间的关系。方法应用逆转录多聚酶链反应（ＲＴ－ＰＣＲ）对２６例术前未作治疗的肝门部胆管癌，１２例正常胆管组织的ｍｄｒ－１ ｍＲＮＡ表达进行了检测，并与癌组织的分化程度，病理类型、部位、浸润转移作对比研究。结果：２６例肝门部胆管癌组织中ｍｄｒ－１ ｍＲＮＡ的阳性表达率为６５．４％（１７／２６），正常胆管     

Fas,P-糖蛋白和多药耐药1基因在急性白血病的表达及相关性的临床研究

 目的 探讨Fas,P-糖蛋白（P-gp）、多药耐药1（mdr1）基因在急性白血病（AL）的表达及三者在多药耐药（MDR）中相关性的临床研究。方法 对59例初发AL患者,应用流式细胞仪和半定量RT-PCR法分别检测治疗前后Fas和P-gp及mdr1表达情况。结果 Fas在不同类型AL表达有差异;Fas+与P-gp+呈显著负相关,Fas+与mdr1+呈负相关,P-gp+与mdr1+呈显著正相关;COX分析结果显示：Fas和mdr1有判断预后价值;Fas+与Fas-组、P-gp+与P-gp-组、mdr1+与mdr1-组的CR率差异均有统计学意义。结论 Fas高表达可能是AL预后良好因素;P-gp,mdr1高表达为预后不良重要因素。     

原发性肝细胞癌肺耐药蛋白LRP基因表达及其意义

Objective: To study the multidrug resistance (MDR) mechanism of lung resistance protein (LRP) gene in hepatocellular carcinoma (HCC), and the relations among the expression of the LRP gene and clinicopathologic features, the influence of -fetoprotein (AFP), and prognosis of patients who received adjuvant chemotherapy after resection of HCC. Methods: The expression of the LRP gene encoding LRP and mRNA LRP was detected in tissues from 54 untreated patients with HCC, adjacent tissues from 24 patients with HCC and archival paraffin-embedded tissues from 12 patients with posthepatitic cirrhosis. The relationship between the LRP gene expression and the change of AFP level was analyzed in the 24 postoperative HCC patients whose AFP was measured after 2 weeks. All of the HCC patients were followed up. Results: The percentage of positive expression of LRP and mRNA LRP in the 3 tissues was 61.1%, 33.3%, 16.7%, and 75.9%, 37.5%, 33.3% respectively. There was significant difference between the untreated HCC tissue and other tissues (P<0.05). No difference existed between the LRP gene expression and clinicopathologic findings, age, sex, and tumor size (P>0.05), but the expression was related to the degree of differentiation of HCC (P<0.05). The effective rate of AFP in the LRP gene positive expression group or in postoperative chemotherapeutic patients was very lower than that in the negative group (P<0.05). Although the mean survival time of postoperative HCC patients in negative LRP gene expression group was longer than that of positive group, there was no difference between them (P<0.05). Conclusion: LRP gene expression is related to MDR of HCC and initiates the intrinsic MDR. Detection of LRP gene expression is of great guiding significance in accessing chemotherapeutic resistance of HCC. As an index to chemotherapy of HCC, detection of LRP expression provides evidence for making individual chemotherapeutic treatment,and reversing MDR in HCC. Although LRP gene expression correlates with the tumor differential degree (P<0.05), it perhaps does not relate with the prognosis of HCC patients.     

Expression and clinical significance of multidrug resistance proteins in brain tumors

Background

To investigate the mechanisms of multidrug resistance of brain tumors, to identify the site of cellular expression of P-gp in human brains in situ and to morphologically determine whether an association may exist between P-gp and caveolin-1.

Methods

Immunohistochemistry was used to detect the expression and location of P-glycoprotein (P-gp), Multidrug resistance-associated protein (MDR), Lung resistance-related protein (LRP), Topoisomerase II (Topo II) and Glutathione-S-π (GST-π) in 30 patient tumor tissues and 5 normal brain tissues. The sections were subjected to double labeling for P-gp (TRITC labeled) and caveolin-1 (FITC labeled). The location and characteristics of expression of the two proteins in the blood brain barrier(BBB) was observed using a laser scanning microscope.

Results

High expression of P-gp was detected in vessel walls and the tissue surrounding the vessels. However, expression of P-gp was low in tumor cells. The expression of the other 4 multidrug resistance proteins was not observed in the vessel walls. Laser scanning microscopy showed P-gp and caveolin-1 co-expression: the two proteins co-localized either in the luminal endothelial compartment or at the border of the luminal/abluminal compartments.

Conclusion

Chemotherapeutics drugs are interrupted in the end-feet of neuroepithelial cells of the BBB by P-gp, which weakens the chemotherapeutic effect. P-gp marks the BBB, and the transporter is localized in the luminal endothelial compartment where it co-localizes with caveolin-1.