大肠癌原发灶及粪便标本中基因mRNA表达及其临床意义

目的通过对COX-2,CK-20,CD44v6mRNA在结直肠原发灶及病人粪便中的表达,证实粪便中COX-2,CK-20,CD44v6mRNA表达来源于原发灶肿瘤细胞的脱落所致,为临床结直肠癌筛查选择合适的多基因标记物联合检测指标。方法对38例行结肠癌根治术的病人,原发灶及术前收集的粪便标本采用RT-PCR技术对COX-2,CK-20,CD44v6mRNA表达进行检查。结果研究发现,COX-2mRNA在原发灶及粪便中同时阳性表达率为44.7%(17/38),同时阴性表达率为39.5%(15/38),同源性可达84.2%(32/38);CK-20有44.7%(17/38)同时阳性表达,36.8%(14/38)同时无表达,同源性为81.5%(31/38);CD44v6mRNA有50%(19/38)同时阳性表达,44.7%(17/38)同时无表达,同源性为94.7%(36/38),同时性表达差异具有显著性,P值均小于0.001。结论粪便COX-2,CK-20,CD44v6mRNA表达与大肠癌原发灶中的表达具有同源性,说明粪便中COX-2,CK-20,CD44v6mRNA来源于原发灶肿瘤细胞的脱落。     

A panel of three serum microRNA can be used as potential diagnostic biomarkers for nasopharyngeal carcinoma

BackgroundNasopharyngeal carcinoma is cancer with unique epidemiological characteristics, showing obvious ethnicity, gender, and geographical prevalence. More and more evidence shows that microRNAs are stable in serum and are specific to different tumor types. Therefore, miRNA is a new non‐invasive biomarker for cancer detection.MethodsThe experiment is divided into three stages, namely, the screening stage, the training stage, and the verification stage. We took 54 patients with nasopharyngeal carcinoma and 108 healthy controls as the research objects. We use the receiver‐operating characteristic (ROC) curve and area under the ROC curve (AUC) to evaluate the diagnostic value of miRNA. Finally, a three‐miRNA panel with high diagnostic efficiency was constructed. In addition, we conducted biological information analysis of these miRNAs to explore their functions.ResultsIn NPC patients, the expression of five serum miRNAs (miR‐29c‐3p, miR‐143‐5p, miR‐150‐5p, miR‐145‐3p, and miR‐205‐5p) is significantly dysregulated. Among them, the diagnostic value of these three miRNAs (miR‐29c‐3p, AUC = 0.702; miR‐143‐5p, AUC = 0.733; and miR‐205‐5p, AUC = 787) is more prominent. The diagnostic panel constructed by them has a higher diagnostic value (AUC = 0.902). Through the analysis of the TCGA data set, the target gene of the three‐miRNA panel may be KLF7, NRG1, SH3BGRL2, and SYNPO2.ConclusionThe three‐miRNA panel (miR‐29c‐3p, miR‐143‐5p, and miR‐205‐5p) may become a novel non‐invasive biological marker for nasopharyngeal cancer screening.     

上皮钙黏着蛋白mRNA表达与肺癌的相关性研究

目的 探讨E-cadherin(上皮钙黏着蛋白,E-cad)mRNA与肺癌及其病理类型之间的相关性.方法 用巢式RT-PCR检测50例肺癌患者的癌、癌旁和远癌肺组织及5例肺良性对照标本中的E-cad mRNA的相对表达量.结果 癌组织1.33±0.53、癌旁组织1.65±0.60、远癌组织2.01±0.46、非肺癌对照肺组织2.09±0.09,经方差分析癌、癌旁和远癌组织差异有统计学意义(F=18.810,P<0.01),但远癌组织与非肺癌对照肺组织之间的差异无统计学意义(P>0.05).肺鳞癌、腺癌及腺鳞癌的癌组织、癌旁组织和远癌组织中E-cad mRNA水平差异无统计学意义(P>0.05).结论 E-cad mRNA表达减少或缺失与肺癌的发生和发展有相关性,与肺癌的组织类型无关.     

非小细胞肺癌患者检测血液 CEA mRNA和CK19 mRNA的意义

目的评价CEA mRNA和CK19 mRNA指标监测非小细胞肺癌(NSCLC)血液转移和评估预后的应用价值。方法以97例NSCLC患者、51例非肿瘤性呼吸系统疾病患者(疾病对照组)和36例健康人(健康对照组)为对象,应用逆转录-套式-聚合酶链反应检测CEA mRNA和CK19 mRNA。结果NSCLC组血液CEA mRNA和CK19 mRNA的阳性结果显著性高于两个对照组(P<0.01),且CEA mRNA的特异性和敏感性也高于CK19 mRNA。指标阴性组患者的24个月生存例数明显多于阳性组(P<0.01~<0.05)。结论检测血液CEA mRNA和/或CK19 mRNA有助于发现NSCLC血液转移,也有助于预后的评估,其中CEA mRNA的敏感性高于CK19 mRNA(P<0.01)。     

Lunx mRNA的表达与非小细胞肺癌外周血微转移的相关性研究

目的 研究Lunx mRNA的表达与非小细胞肺癌外周血微转移的相关性.方法 采用RT-PCR(逆转录PCR)技术分别对实验组及对照组外周血Lunx mRNA的表达进行检测分析.结果 NSCLC 患者外周血微转移与病理类型、细胞分化程度及 TNM 分期均存在密切关系(P＜0.05);NSCLC患者外周血微转移与年龄、性别、吸烟史、原发肿瘤部位无密切关系(P＞0.05).结论 Lunx mRNA是一种检测 NSCLC 外周血微转移新型的、较好的特异性标记物,对判断病程发展和指导临床治疗具有重要意义.     

Glucosamine‐phosphate N‐acetyltransferase 1 and its DNA methylation can be biomarkers for the diagnosis and prognosis of lung cancer

ObjectiveLung cancer ranking high in the cancer‐related list has long perplexed patients, in which glucosamine‐phosphate N‐acetyltransferase 1 (GNPNAT1) is found to be highly expressed. Besides, DNA methylation is perceived as a biomarker to assess the prognosis of patients with various cancers. However, the correlation between GNPNAT1 and DNA methylation and the role of GNPNAT1 in lung cancer remain vague.MethodsPrincipal component analysis (PCA), heatmap, volcano map, Venn diagram, gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to screen out the candidate genes. The viability, migration, and invasion of lung cancer cells were detected by CCK‐8 and Transwell assays. An xenograft tumor mouse model was established. The relative expressions of GNPNAT1, E‐cadherin, vimentin, Matrix metalloproteinase‐2 (MMP‐2), tissue inhibitor of metalloproteinase‐2 (TIMP‐2), E2F1, and cyclin D1 in cells or xenograft tumor tissues were quantified by Western blot, RT‐qPCR, or immunohistochemistry assay.ResultsGNPNAT1 was screened as the research object. GNPNAT1 methylation was downregulated, while GNPNAT1 expression was upregulated in lung cancer tissues. The methylation and mRNA levels of GNPNAT1 were correlated with the patient prognosis. GNPNAT1 increased cell viability, migration and invasion, and promoted the xenograft tumor volume and weight, whereas shGNPNAT1 acted oppositely. Moreover, expressions of Vimentin, MMP‐2, E2F1, and cyclin D1 were increased, but E‐cadherin and TIMP‐2 expressions were decreased by overexpressed GNPNAT1, whilst GNPNAT1 knockdown ran conversely.ConclusionGNPNAT1 and methylated GNPNAT1 coverage are biomarkers for the diagnosis and prognosis of lung cancer.     

Plasma SNORD42B and SNORD111 as potential biomarkers for early diagnosis of non‐small cell lung cancer

BackgroundNon‐small‐cell lung cancer (NSCLC) still occupied the leading reason of cancer death due to lack of availability of early detection. This study aimed to identify the effective biomarkers for the early‐stage NSCLC diagnostics based on plasma snoRNAs.Materials and MethodsThe differential snoRNAs between lung cancer patients and healthy donors were analyzed using the SNORic and TCGA databases. SNORD42B and SNORD111 were screened out and further verified in 48 FFPE NSCLC and adjacent normal tissues, as well as in plasma from 165 NSCLC patients and 118 health donors using qRT‐PCR. Next, their diagnostic efficiency, as well as combined with carcinoembryonic antigen (CEA), was obtained by the analysis of receiver operating characteristic (ROC).ResultsWe first screened out 47 top differential snoRNAs, among which the top 10 upregulated snoRNAs in LUAD were U44, U75, U78, U77, SNORD72, SNORD13, SNORD12B, SCARNA5, U80, SNORD41, and in LUSC were U44, U75, U78, SNORD41, SNORD111, SNORA56, U17a, SNORD35A, SNORD32A, SNORA71D. SNORD42B and SNORD111 was significantly increased not only in tumor tissues but also in plasma from NSCLC and early‐stage NSCLC patients. They were capable to act as promising biomarkers for NSCLC and early‐stage NSCLC diagnosis. Moreover, CEA diagnostic efficiency for early‐stage NSCLC was significantly improved when combined with these two plasma snoRNAs.ConclusionSNORD42B and SNORD111 could act as the potential and non‐invasive diagnostic biomarkers for NSCLC and early‐stage NSCLC.     

DNA from blood samples can be used to genotype patients who have recently received a transfusion

BACKGROUND: The use of hemagglutination to phenotype red cells from recently transfused patients or of red cells that are coated with IgG can be time-consuming and difficult to interpret. Because the molecular bases of many blood group antigens are known, it was investigated whether polymerase chain reaction (PCR) analysis of DNA, from white cells in blood from transfused patients, could be used to predict the blood group antigen profile of a patient. STUDY DESIGN AND METHODS: To prevent problems arising from potentially poor-quality DNA in clinical samples, primers that flanked the polymorphism of interest and that replicated a relatively short PCR amplicon were used. The PCR products, with or without digestion with the appropriate restriction enzyme, were analyzed on gels. Samples were collected from 60 patients who had received from 2 to 50 units of RBCs in the 7 days before sample collection. RBCs from some of these patients were coated with IgG. Analyses for RHD/non-D, RHE/RHe, KEL1/KEL2, FYA/FYB, FY-GATA, JKA/JKB, and GYPA M/N were performed by using assays that had been validated with DNA prepared from untransfused volunteers of known phenotype. The genotyping assays were performed without knowledge of the expected result. RESULTS: The predicted genotype after analysis of the 60 patient samples was that expected from the results of phenotyping. In all cases, the molecular analysis gave a single result; no evidence of chimerism was obtained. CONCLUSION: In each case, the molecular genotype results were in agreement with the blood group antigen as determined by historical phenotyping, phenotyping after hypotonic washing, detection of alloantibodies in the patient's serum, or elution of alloantibody(ies). Under the conditions of these assays, reliable determination of a blood group allele can be made by PCR-restriction fragment length polymorphism testing.     

ProGRP: a new biomarker for small cell lung cancer

Progastrin-releasing peptide (ProGRP) is a recently identified biomarker of small cell lung cancer (SCLC), a disorder of neuroendocrine tissue differentiation. The upper normal limit of ProGRP in the circulation is 50 pg/ml. Impaired glomerular filtration tends to increase circulating levels and confound the tumor marker significance of modestly elevated values. Excluding patients with renal failure, circulating levels did not exceed 80 pg/ml in benign disease (3% of cases in excess of the upper normal limit) or 120 pg/ml in malignancy other than lung cancer and neuroendocrine tumors (5% of cases in excess of the upper limit). ProGRP serum levels are clearly related to the lung cancer histological type with significantly higher levels observed in SCLC than in nonsmall cell lung cancer (NSCLC). Circulating ProGRP in excess of 120 mg/ml was found in only 4% of cases of NSCLC with another 22% presenting with modestly elevated levels in excess of the upper normal limit. By contrast, abnormal ProGRP results are found in 60-70% and in 75-90% of SCLC patients with local and extensive disease, respectively. ProGRP is a more sensitive biomarker than is neuron-specific enolase (NSE) for SCLC, but thus far has not been found in multivariate analysis to have independent prognostic significance. Preliminary studies suggest ProGRP will have utility in conjunction with NSE in monitoring the therapy of established SCLC.     

Belagenpumatucel-L for the treatment of non-small cell lung cancer

Introduction: Immunotherapy has become a promising approach for the treatment of NSCLC. In order to stimulate the host immune system against tumour antigens, several cancer vaccines have been generated and evaluated. Belagenpumatucel-L is a whole tumour cell vaccine expressing the antisense strand of the TGF-β2 gene.

Areas covered: The purpose of this article is to review the most relevant findings of clinical trials testing belagenpumatucel-L in advanced NSCLC patients.

Expert opinion: Although the Phase III trial investigating belagenpumatucel-L in stage III/IV patients did not meet its primary end point, a survival benefit was observed in several subgroups of patients. Further studies are needed in order to select patients who may benefit from this vaccine.     

Circular RNA hsa_circ_0000567 can be used as a promising diagnostic biomarker for human colorectal cancer

Background

Recent studies have revealed that circular RNAs are involved in the biological process of some kinds of human cancers. However, little is known about their diagnostic values and functions in colorectal cancer (CRC).

Methods

The expression levels of hsa_circ_0000567 in 102 paired CRC tissues and adjacent noncancerous tissues, 5 CRC cell lines, and a normal colorectal epithelial cell line were detected by quantitative real‐time polymerase chain reaction (qRT‐PCR). The correlations between hsa_circ_0000567 expression levels and the clinicopathological factors of patients with CRC were analyzed. Furthermore, the loss‐of‐function assay was performed to investigate the functions of hsa_circ_0000567 in vitro. Finally, a receiver operating characteristic (ROC) curve was established to evaluate the diagnostic value of hsa_circ_0000567.

Results

Hsa_circ_0000567 expression was significantly downregulated in CRC tissues and CRC cell lines. In addition, the decreased hsa_circ_0000567 expression in CRC was negatively correlated with tumor size (P = .011), lymph metastasis (P = .003), distal metastasis (P < .0001), and tumor–node–metastasis (TNM) stage (P = .003) in CRC. Moreover, knockdown of hsa_circ_0000567 promoted CRC cells proliferation and migration in vitro. Importantly, the area under the ROC curve (AUC) was 0.8653, which indicates hsa_circ_0000567 can serve as a diagnostic biomarker.

Conclusion

Hsa_circ_0000567 may be a novel suppressor and a potential diagnosis biomarker in CRC.

     

LunX mRNA联合CEA、SCCA、CYFRA21—1三项肿瘤标志物诊断非小细胞肺癌的临床应用价值

目的 探讨外周血LunX mRNA联合CEA、SCCA、 CYFRA21-1三项肿瘤标志物检测在非小细胞肺癌(NSCLC)患者临床诊断中的价值.方法 应用实时荧光定量RT-PCR技术和化学发光免疫分析法,分别对NSCLC、乳腺癌、胃癌、肺炎患者及健康体检人群外周血中LunX mRNA和三项肿瘤标志物水平进行检测.结果 NSCLC患者外周血中检测到LunX mRNA,阳性表达率为78.6%(44/56),在乳腺癌、胃癌、肺炎患者健康体检人群外周血中LunX mRNA检测为阴性,但是其他三项肿瘤标志物在NSCLC、乳腺癌、胃癌外周血中检测为阳性,在肺炎患者和健康体检者中检测为阴性.外周血中LunX mRNA表达水平及其阳性表达率与NSCLC患者TNM病理分期密切相关(P＜0.05).结论 NSCLC患者外周血中LunX mRNA表达特异性高, LunX mRNA联合三项肿瘤标志物检测对NSCLC临床诊断提供重要参考依据.     

非小细胞肺癌预后相关评价标志物的筛选

目的利用生物芯片技术和非小细胞肺癌（non-small cell lung cancer,NSCLC）患者血清及随访结果,筛选出新型NSCLC高敏感性和特异性的血清自身免疫抗体作为分子标志物用于NSCLC的预后评价。方法（1）提取NSCLC组织总mRNA构建T7噬菌体cDNA文库;（2）用NSCLC预后良好和不良患者血清对T7文库进行生物淘洗;（3）构建蛋白芯片,分别用预后良好和不良患者血清孵育芯片进行CyS／Cy3双荧光标记并分析芯片结果;（4）对挑选出的标志物进行测序及分析。结果筛选得到最佳评价组合含6个NSCLC预后相关标志物,其联合诊断准确率80．7％,敏感性85．3％,特异性73．9％,AUC为0．825;测序及BLAST分析显示,abl—interactor 1、pleiotrophin、surfactantprotein B 3个标志物是已知癌细胞转移和预后相关分子。结论成功筛选得到1个含有6个标志物的最佳评价组合,可对NSCLC预后进行较准确诊断。     

Emepepimut-S for non-small cell lung cancer

Introduction: Immunotherapy as a possible therapeutic option for cancer has been of great importance due to the innovative development of vaccines. Various molecules have been tested and emepepimut-S (Biomira Liposomal Peptide 25 (BLP 25)) has emerged as an option, particularly in lung cancer.

Areas covered: A PubMed literature and ClinicalTrials.gov search was conducted using the terms: emepepimut, BLP25, NSCLC, cancer immunotherapy, cancer vaccine and MUC1. This review covers how emepepimut-S acts against the mucin 1 (MUC1) tumor-associated antigen producing a cellular immune response against the cells that express MUC1 and the most important clinical data available that led to the ongoing Phase III trial.

Expert opinion: The results obtained in the Phase I/II trials are promising, showing a favorable toxicity with a benefit in survival in NSCLC patients. As future trials develop, demonstration of the long-term survival benefit, understanding of the various mechanisms of immune response initiated by the drug and the selection of patients that will highly benefit from the immunotherapy will be elucidated. The safety and extension in survival makes emepepimut-S a very interesting drug and could, therefore, offer a possibility of treatment and maintenance, particularly for good performance status patients with locally advanced NSCLC.     

裸鼠颅内小细胞肺癌移植瘤模型的建立

目的：建立裸鼠颅内小细胞肺癌（SCLC）移植瘤模型,为今后开展SCLC相关实验研究提供动物模型。方法培养LTEP/P人SCLC细胞株,传代3～4代,收集细胞,并制成1×108/ml 的细胞悬液共1 ml。14只裸鼠麻醉后,用牙科钻在裸鼠颅脑钻一小孔,然后用10μl微量注射器抽取3μl肿瘤细胞悬液,利用立体定位仪缓慢接种于BALB/C SPF裸鼠颅内,放置入SPF环境内继续饲养,观察7 d后脱颈处死,取完整脑组织,行病理检测。结果所有裸鼠在术后1h内逐渐苏醒并恢复活动和饮食,第5天出现活动、饮食减少,第7天出现神软、萎靡、反应迟钝等症状,并死亡1只。病理检测证实所有裸鼠脑组织内可见SCLC细胞。结论通过立体定位仪向裸鼠颅内接种SCLC细胞能建立稳定可靠的裸鼠颅内SCLC移植瘤模型。     

联合化疗方案治疗非小细胞肺癌对照研究

目的：探讨联合化疗方案治疗非小细胞肺癌的临床效果。方法将60例非小细胞肺癌患者按照随机数字表法分为两组，每组30例，两组均常规予以吉西他滨、顺铂治疗，实验组在此基础上联合索拉非尼治疗，随访3 a。采用实体肿瘤疗效评价标准评估两组近期、远期治疗效果，随时记录不良反应发生状况。结果治疗后两组近期、远期治疗效果比较差异均无显著性(P >0.05)，实验组除手足综合征发生率显著高于对照组(P <0.05)外，其他不良反应发生率与对照组比较差异均无显著性(P >0.05)。结论吉西他滨、顺铂联合索拉非尼联合化疗方案对改善非小细胞肺癌患者的总生存期和远期存活率效果不明显，临床应根据患者病理特点和生物标记物进行个体化治疗。     

超声支气管镜引导下针吸活检术结合免疫组化及基因学检测在肺癌诊治中的应用*

目的评价超声支气管镜引导下针吸活检术(EBUS-TBNA)结合免疫组化技术和基因学检测在肺癌的诊断及治疗中的应用价值。方法回顾性分析从2013年5月-2015年10月共55例初诊的临床诊断纵隔占位或疑为肺癌伴纵膈或肺门淋巴结转移的患者,为明确诊断行EBUS-TBNA检查,必要时联合免疫组化检测进一步完善诊断及分型,并对部分肺癌患者行表皮生长因子受体(EGFR)或间变性淋巴瘤激酶(ALK)检测以明确分子分型从而为靶向治疗提供依据。结果 55例患者中,经EBUS-TBNA确诊为肺癌37例,肺转移癌3例,软组织肉瘤1例,转移性淋巴结腺癌1例,淋巴瘤1例,结核病4例,慢性炎症1例,7例经EBUS-TBNA未能明确诊断。EBUS-TBNA在该组肺癌诊断中的敏感性及准确性分别为92.5%(37/40)和94.5%(52/55)。29例EBUS-TBNA组织标本依赖免疫组化检测证实为肺癌且得到明确分型,其中17例肺腺癌,6例肺鳞癌,6例小细胞肺癌。6例EBUS-TBNA组织标本行EGFR基因检测,其中1例同时行ALK基因检测,基因检测结果示EGFR基因突变型4例,ALK基因无融合1例。所有病例无1例严重并发症发生。结论 EBUS-TBNA诊断肺癌具有良好价值,且安全性较好,结合免疫组织组化检测及基因突变检测对肺癌的诊断及指导治疗有突出的价值。     

Management of the elderly patient with advanced non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the most lethal of the common solid malignancies. It is predominantly a disease of the elderly with the median age at diagnosis 68 years. Unfortunately, the majority of patients present with advanced disease whereby palliation is the primary aim of treatment. The elderly have a long history of undertreatment and non-inclusion in clinical trials with regard to cancer. Elderly-specific studies demonstrate that chemotherapy provides both a survival and quality-of-life benefit in advanced NSCLC. Increasing emphasis is placed on the objective assessment of fitness for chemotherapy and the integration of molecularly targeted agents into treatment paradigms.     

衣壳蛋白和甲基转移酶对戊型肝炎病毒的基因分型

目的探讨人戊型肝炎病毒(HEV)的部分编码区序列能否作为其基因组分型的依据。方法从GenBank中查找并下载全基因组测序完整的能感染人类的HEV基因组及蛋白序列共35株,用Clustal W程序对35株HEV全序列以及部分编码区序列进行多序列比对,并用treev 32绘制基因进化树,研究其进化关系。结果HEV的基因组全长7.5kb;而衣壳蛋白和甲基转移酶的编码序列分别为1 982bp和545bp,这2种蛋白的多序列比对和进化分析结果与完整基因组的分型结果相符。结论衣壳蛋白和甲基转移酶可作为HEV的分型依据,且是一种更简便、快捷的方法。