应用PCR-SSCP银染法检测胰腺癌组织K-RAS基因突变

我们用PCR-SSCP银染技术检测了29例胰腺癌、8例胰岛细胞瘤和9例慢性胰腺炎石蜡包埋组织标本中K-Ras基因外显子1,发现有17例胰腺癌标本存在突变,突变率为58.6％,而无一例胰岛细胞瘤及慢性胰腺炎标本有突变发生。胰腺癌标本突变率与肿瘤分化程度、病人的性别及年龄无关,提示K-Ras基因突变普遍存在于胰腺癌病人中,对此突变的检则可用于胰腺癌的鉴别诊断。PCR-SSCP(聚合酶链反应-单链构型多态分析)银染方法简便、快速。可广泛用于基因突变的检测。     

胰腺癌相关生物活性标志物研究

肿瘤相关生物活性标志物主要是在肿瘤发生发展过程中出现的基因变化以及合成和分泌的异常蛋白质,并可以用于诊断和鉴别肿瘤的物质。临床中胰腺癌发生是一个复杂的多基因改变以及多种蛋白表达异常的结果,这可导致相应基因突变以及蛋白的高表达,同时部分指标可以成为胰腺癌相关生物活性标志物。     

精准医疗时代胰腺癌个体化类器官研究现状与应用前景

目的总结精准医疗时代背景下胰腺癌个体化类器官(organoids)研究现状及其潜在的临床应用前景。方法检索Pub Med数据库获取相关资料,就胰腺癌个体化类器官特征及其在促进胰腺癌精准治疗中的应用进行综述。结果胰腺癌患者个体化类器官模型系统作为测试肿瘤药物和个体化肿瘤疗法的新型平台,利于致癌基因建模、靶基因发现和个体化药物敏感性检测,可用于肿瘤精准治疗。结论个体化肿瘤类器官可用于指导胰腺癌的精准治疗。     

外周血浆K-ras基因突变的检测在胰腺癌诊断中的作用

目的 探讨检测K－ras基因突变在胰腺癌诊断中的作用。方法 收集15例胰腺癌,33例非胰腺癌病人的外周血标本,提取血浆DNA。采用限制性酶切片段长度多态性- 多聚酶链反应（RFLP－PCR）方法检测突变的K－ras基因,直接测序方法测出K－ras基因的突变类型。结果 胰腺吕病人外周血浆K-ras基因突变率为73％（11／15）,非胰腺癌组中有2例胰头肿块型慢性胰炎患者发生K-ras基因突变,其余均为阴性;血浆K－ras基因突变与肿瘤部位、大小及分期无关。结论 检测外周血浆K－ras基因突变可用于胰腺癌的辅助诊断和胰腺癌高危人群的筛选。     

应用聚合酶链反应-单链构型多态分析和银染法检测胰腺癌组织K-Ras基因突变

用聚合酶链反应—单链构型多态分析(PCR-SSCP)和银染技术检测了29例胰腺癌、8例胰岛细胞瘤和9例慢性胰腺炎石蜡包埋组织标本中K-Ras基因外显子1,发现有17例胰腺癌标本存在突变,突变率为58.6％,而无1例胰岛细胞瘤及慢性胰腺炎标本发生突变。胰腺癌标本突变率与肿瘤分化程度、病人的性别及年龄无关,提示胰腺癌普遍存在K-Ras基因突变,检测此突变对胰腺癌的鉴别诊断有一定临床意义。PCR-SSCP银染方法简便、快速,可广泛用于基因突变的检测。     

腹腔镜在胰腺外科中的应用进展

腹腔镜技术的出现与日趋完善,是对胰腺外科的诊断、治疗的重要补充。利用腹腔镜对胰腺癌进行术前诊断和分期。对可以切除的胰腺癌行胰十二指肠切除术和胰体尾切除术,而对已无法切除的胰腺癌可应用腹腔镜行姑息手术治疗;腹腔镜可用于急性胰腺炎和胰腺囊肿引流的诊断、治疗。     

胰腺癌基因诊治的新进展

胰腺癌基因诊断和治疗的进展有助于改善胰腺癌的预后。人们在研究中不断发现新的癌基因参与胰腺癌的发生和发展 ,同时还寻求置换有正常功能的基因或增补缺陷基因的方法治疗胰腺癌。可以预见 ,基因诊断和治疗作为一种全新的胰腺癌诊治手段 ,将一定程度改变人类疾病治疗的进程。一、基因诊断随着分子生物学技术的进展 ,现在我们可以直接检测和鉴定缺陷基因 ,使胰腺癌的诊断从传统的表型诊断上升到基因诊断。现已证实胰腺癌的发生和发展与抑癌基因、原癌基因、ＤＮＡ错配修复基因及雌激素诱导基因等有关。1.抑癌基因 :已知至少 10种抑癌基因与…     

胰腺癌基因研究

胰腺癌基因研究南通医学院肝胆胰外科研究室（２２６００１）蔡斌综述，陈玉泉审校胰腺癌的发病率逐年增高，但有关胰腺癌的诊断和治疗仍未取得突破性进展。随着分子生物学的发展，胰腺癌的基因研究也受到许多学者的重视。胰腺癌有关的基因胰腺癌有关的基因包括Ｃ─ｋｉ─...     

寡核苷酸芯片技术在胰腺癌诊断中的应用

目的 探讨基因突变寡核苷酸芯片技术检测K-ras基因与p53基因突变在胰腺癌诊断中的价值。方法应用芯片技术检测25例胰腺癌和5例慢性胰腺炎患者外周血浆游离DNA和组织标本中K-ras基因与p53基因突变。结果胰腺癌病人外周血血浆游离DNA中K-ras基因突变率为52.0％,p53基因突变率为20.0％;联合检测阳性率为60.0％,肿瘤组织标本中K-ras基因突变率为76.0％,p53基因突变率为32.0％,联合检测阳性率为92.0％。结论寡核苷酸芯片技术能够为胰腺癌的相关基因分析提供可靠的数据,检测K-ras基因突变有助于胰腺癌早期诊断,联合检测p53基因突变可提高诊断的敏感性和特异性。     

胰腺癌的肿瘤标志物和免疫基因治疗研究进展

胰腺癌由于恶性程度高、手术切除率低、早期诊断困难 ,治疗的效果不尽人意。近年来 ,随着对肿瘤的发生、发展和转移机制认识的不断深入 ,尤其是肿瘤分子生物学与免疫学的进展 ,关于肿瘤相关的免疫调节与应答、癌变、转移以及癌基因和抑癌基因之间的作用机制的进一步了解 ,许多用于胰腺癌早期诊断的血清及基因标志物被发现 ,不少的免疫基因治疗手段已经进入临床试验[1,2 ] ,为胰腺癌的诊断和治疗提供了新的希望。胰腺癌的肿瘤标志物胰腺癌传统的肿瘤标志物通常指由肿瘤组织自身合成、分泌的某些抗原、激素及酶类等 ,但是 ,随着分子生物学技术…     

生物信息挖掘指导胰腺癌诊治的现状与展望

胰腺癌作为目前最致命的人类肿瘤类型,其生物学上的侵袭性、早期发现策略的空白和有效治疗手段的缺乏是导致胰腺癌生存率低的主要原因。深入了解胰腺癌的发生发展机制,早期诊断胰腺癌,有效预测胰腺癌的复发及预后,探索关键的治疗靶点是新时代胰腺癌研究的重中之重。大规模的基因组研究对了解胰腺癌关键驱动基因奠定了一定的基础。在后基因组时代,通过对胰腺癌患者的肿瘤组织和体液进行蛋白测序,将逐渐开创一个精准诊治胰腺癌的新时代。目前全球范围以胰腺外科为主的胰腺癌多学科诊疗(MDT)医疗团队利用癌症基因组图谱(TCGA)、高通量基因表达数据库(GEO)、国际癌症基因组联盟(ICGC)等大规模公开数据库,结合各中心小规模测序数据,逐渐揭开胰腺癌在不同生物过程中的核心肿瘤标志物,进而为临床转化治疗提供理论基础及指导。     

DNA甲基化修饰在胰腺癌中的研究进展

胰腺癌的发病机制、早期诊断及治疗一直是医学界关注的热点与重点。近年来多项研究表明表观遗传学对肿瘤的发生发展发挥着重要的作用，其中DNA甲基化最常见。胰腺癌的发生发展与其相关癌基因或抑癌基因由于DNA甲基化水平变化引起的异常激活或抑制有关。DNA甲基化可能在体细胞突变前发生，且贯穿肿瘤的全过程，因此在肿瘤的诊断、治疗和预防中被广泛研究。笔者对DNA甲基化的概念、作用方式、在胰腺癌发生发展中所起的作用以及胰腺癌诊断和治疗上的前景作一综述，以期对胰腺癌未来的研究提供一定的参考。     

The role of epigenetic alterations in pancreatic cancer

The past several years have witnessed an explosive increase in our knowledge about epigenetic features in human cancers. It has become apparent that pancreatic cancer is an epigenetic disease, as it is a genetic disease, characterized by widespread and profound alterations in DNA methylation. The introduction of genome-wide screening techniques has accelerated the discovery of a growing list of genes with abnormal methylation patterns in pancreatic cancer, and some of these epigenetic events play a role in the neoplastic process. The detection and quantification of DNA methylation alterations in pancreatic juice is likely a promising tool for the diagnosis of pancreatic cancer. The potential reversibility of epigenetic changes in genes involved in tumor progression makes them attractive therapeutic targets, but the efficacy of epigenetic therapies in pancreatic cancer, such as the use of DNA methylation inhibitors, remains undetermined. In this review, we briefly summarize recent research findings in the field of pancreatic cancer epigenetics and discuss their biological and clinical implications.     

基于机器学习的胰腺癌特征基因筛选初步研究

背景与目的 胰腺癌是一种难治的癌症,90%以上的患者在诊断后1年内死亡。胰腺癌病变组织和正常组织之间存在差异表达基因（DEGs）可能与胰腺癌的发生和发展密切相关。本研究运用机器学习方法对胰腺癌DEGs进行筛选,以期为研究该病的发生机制提供依据。方法 从公共基因GEO数据库中筛选胰腺癌基因表达谱,使用线性回归模型软件包Limma对不同组的芯片进行差异性计算,归一化;使用R语言获得DEGs,对筛选出来的DEGs特征选择方法进一步进行筛选;基于获得的核心DEGs,采用AdaBoost和Bagging算法分别构建胰腺癌预测模型。用DAVID 网站对核心DEGs进行GO功能分析和KEGG通路富集分析,再用STRING网站及Cytscape软件对核心DEGs进行蛋白-蛋白相互作用（PPI）网络分析,最后用GEPIA网站对预后相关的核心DEGs行生存分析。结果 通过特征筛选,得到了18个关键的DEGs;以该18个DEGs建立特征子集,结合AdaBoost算法建立了预测模型,预报准确率可以达到92.3%。通过对DEGs的GO和KEGG分析,发现CDK1、CCNA2和CCNB1有间接作用,对胰腺癌的形成和发展有一定的作用。生存分析显示,CDK1（P=0.000 8）、CCNB1（P=0.012）、CSK2（P=0.023）、CKS1B（P=0.001 3）的表达量与患者总生存期（OS）有相关性,它们的表达量越高,患者OS越短。结论 机器学习方法可较好地对胰腺癌特征基因进行筛选,对胰腺癌的诊治及相关的药物开发具有一定意义。     

The molecular genetics of pancreatic ductal carcinoma

Pancreatic ductal carcinoma remains the 4th leading cause of cancer death in both men and women in the United States, with an overall 5-year survival of less than 3%. Over the last decade, significant advances have been made in our understanding of the molecular biology of pancreatic ductal carcinoma, with pancreatic cancer now considered one of the better characterized neoplasms at the genetic level. The advances in the understanding of the molecular genetics of pancreatic cancer initially focused on events that occur in the development and early genetic progression of the disease. This progression has been associated with the accumulation of multiple genetic alterations in various cancer-causing genes, leading to the development of a histological and genetic progression model. In the model, pancreatic cancer develops from non-invasive intraepithelial precursor lesions termed pancreatic intraepithelial neoplasias, with each progressive stage associated with accumulated mutations in oncogenes, tumor-suppressor genes, and mismatch repair genes. Other aspects of the development of pancreatic ductal carcinoma, such as tumor invasion, tumor-stromal interaction, metastasis, and chemotherapeutic resistance are more poorly understood. Recent studies utilizing global gene expression methodologies have provided insight into some of these processes and have allowed for the development of potential tumor markers which could be used for early detection and diagnosis of this difficult disease. In order to improve the survival of patients with pancreatic carcinoma, we need to better understand the fundamental changes that occur in pancreatic ductal carcinoma. The following article reviews the genetic mutations and syndromes known to be associated with pancreatic ductal carcinoma as well as recent advances in the study of global gene expression.     

Molecular markers of pancreatic cancer: development and clinical relevance

Background  The prognosis of pancreatic cancer remains poor, mainly because of its aggressive biological behaviour and late clinical diagnosis, which precludes the application of appropriate curative therapies. Therefore, one of the major goals in clinical pancreatology is to find molecular markers, specific and sensitive enough to make an early and correct diagnosis of pancreatic cancer, before it has disseminated and become untreatable. Objective  This overview article explores the potential utility of current molecular markers for the diagnosis of pancreatic cancer. Results  There is a wide array of serum-based and tissue-based markers for pancreatic cancer. Serum-based molecular markers include CA 19-9, CA 125, M2-PK and secreted proteins. A tissue can be used to test genetic mutations such as K-ras, inactivation of tumour suppressor genes (e.g. p16, p53), mucins, telomerase activity, growth factors, DNA methylation, and global gene expression of cDNA microarrays, mitochondrial mutations and proteomics. None of these markers is currently useful for the detection of early pancreatic cancer. In clinical practice, the most commonly accepted use of CA 19-9 is to assess the prognosis and monitor the response to therapy. Conclusions  Many molecular markers have been proposed for the early diagnosis of PC, but most are not ready to be included as part of the routine diagnostic algorithm because they still lack sensitivity, specificity or reproducibility. CA 19-9 remains the most useful molecular marker for the diagnosis and follow-up of clinically and radiological evident pancreatic cancer.     

中国胰腺癌综合诊治指南(2020版)

近年来,胰腺癌的发病率呈上升趋势,其死亡率居恶性肿瘤的第3位。过去十年,胰腺癌的诊治取得了长足进步,然而国内不同地区水平参差不齐,其诊治现状依然严峻。2018年,中国抗癌协会胰腺癌专业委员会制定了《中国胰腺癌综合诊治指南(2018版)》,以期规范和提高中国胰腺癌诊治水平。2020年,专委会结合过去两年的发展现状制定了《中国胰腺癌综合诊治指南(2020版)》。其更新要点主要体现在以下方面:靶向和免疫治疗取得突破,遗传筛查及基因检测首次全面融入胰腺癌的综合诊治。第8版AJCC-TNM胰腺癌分期系统的实用性和准确性在国内多个中心获得验证,并用于临床实践。术前新辅助治疗成为交界可切除和局部进展期胰腺癌的标准治疗方式,并逐渐应用于可切除胰腺癌,新辅助治疗后手术探查尤为重要。以化疗为基础的系统治疗模式(包括靶向治疗和免疫治疗)进入临床研究,并在晚期胰腺癌中证实了维持治疗模式的临床获益性。多学科、多区域协作诊疗模式在国内广泛普及,并贯穿诊疗全程。国内临床试验的开展和多中心跨区域合作为胰腺癌新药研发和方案优化提供了适合国人的高级别循证医学证据。在新的理念和临床证据支持下,该版指南有望为我国胰腺癌的综合诊治工作提供指导。     

Application of Molecular Biology Studies to Gene Therapy Treatment Strategies

The diagnosis of pancreatic cancer continues to produce fear in both patients and practitioners in large part owing to the likely incurability in all for whom the diagnosis is made. It is this reality that continually motivates the surgical and medical oncologists who endeavor to treat these patients. Currently, the cure rate for pancreatic cancer has improved only minimally, and the overall survival of patients remains dismal, with fewer than 5% of patients alive at 5 years and 92% of these patients dead at 2 years. This current treatment status has stimulated numerous studies endeavoring to understand the diverse mechanisms of cell growth in this tumor. Intensive investigative efforts have produced the understanding of new tumor suppressor genes such as DPC4 and an increasing understanding of tyrosine kinase receptors and signal transduction and their regulation of programmed cell death (apoptosis). Detection of these numerous genetic defects may give new insights and understanding of the highly chemo- and radioresistant nature of pancreatic cancer. These same findings also provide the basis for the development of new potential therapies for pancreatic cancer through gene therapy. This paper reviews the significant molecular biologic findings and their influence on the development of gene therapy strategies in the treatment of pancreatic adenocarcinoma.     

微小RNA在胰腺癌中的研究进展

目的总结微小RNA(microRNA,miRNA)与胰腺癌之间关系的研究现状,探讨其表达谱在胰腺癌诊断中的重要作用。方法应用PubMed及CNKI期刊全文数据库检索系统,以"microRNA、胰腺癌"等为关键词,检索2000～2012年的相关文献,共检索到英文文献60篇和中文文献15篇。纳入标准为:miRNA与胰腺癌的基础与临床研究以及miRNA在胰腺癌诊治中的前景。根据纳入标准,纳入31篇文献。结果研究发现,miRNA表达谱如miR-21、miR-34、miR-217、miR-196a、miR-10a、miR-155、miR-221、miR-222、miR-181a、miR-181b、miR-181d、miR-200、let-7等家族成员可作为肿瘤标志物用于胰腺癌与正常胰腺、慢性胰腺炎、胰腺内分泌肿瘤等的诊断和鉴别诊断,预测胰腺癌的预后等。结论胰腺癌中miRNA的表达谱不但与胰腺癌的诊断相关,也为胰腺癌的基因治疗提供了新的研究方向和方法。     

人胰腺癌干细胞差异性基因的表达

目的 分选、鉴定人胰腺癌干细胞,运用基因芯片技术分析其差异性基因的表达.方法 运用流式分选技术分选胰腺癌干细胞(CD24+CD44+ESA+),NOD/SCID鼠移植瘤试验进行肿瘤干细胞特性鉴定.采用Affymetrix U133 plus2.0人类全基因组表达谱芯片对胰腺癌干细胞和非干细胞进行差异基因筛选.结果 分选得到人胰腺癌CD24+CD44+ESA+亚群细胞,占所有细胞的0.8%;5×103个CD24+CD44+ESA+细胞就能成瘤(2/4),而阴性细胞1×105才能成瘤(1/4);CD24+CD44+ESA+具有一定的自我更新和分化能力.基因芯片杂交获得6553(11.99%)条差异基因,胰腺癌干细胞中5255(9.61%)条上调表达,1298(2.37%)条下调表达.其中差异基因涉及细胞凋亡、细胞周期、代谢、细胞线粒体结构和耐药等多个方面.结论 胰腺癌于细胞具有自身特征性基因表达谱,为进一步从干细胞层面研究胰腺癌发病机制及靶向治疗奠定基础.