实时荧光定量RT-PCR对胃癌组织中hPMS2 mRNA 的检测及其临床意义

目的:探讨DNA错配修复基因hPMS2在胃癌组织中的表达水平及其临床意义.方法:应用实时荧光定量逆转录聚合酶链反应(RT-PCR)技术对41例胃癌患者的癌组织、37例癌旁萎缩性胃炎组织、25例慢性萎缩性胃炎组织及20例慢性浅表性胃炎组织中hPMS2 mRNA进行定量检测,以三磷酸甘油醛脱氢酶基因(hGAPDH)为内参照.结果:胃癌组织、癌旁萎缩性胃炎组织、慢性萎缩性胃炎组织及慢性浅表性胃炎组织中hPMS2 mRNA的含量分别是28.33±14.05,16.88±10.67,7.25±8.72和1.78±1.34,四组相比差异有显著性(F=31.82,P=0.00).胃癌组织中hPMS2 mRNA含量明显高于其他三组;癌旁萎缩性胃炎组织中含量明显高于非胃癌患者的胃炎组织,差异均有显著性(P＜0.01);而非胃癌患者的慢性萎缩性胃炎组织中hPMS2 mRNA的含量与慢性浅表性胃炎组织相比差别无显著性.hPMS2 mRNA含量与肿鬃瘤的大小、浸润深度、有无淋巴结转移关系不明显,差异均无显著性.结论:hPMS2基因的异常转录可能与胃癌的发生有关,而与胃癌的发展关系不大.     

Detection of cancer stem cell like circulating tumor cells in peripheral blood of 45 patients with gastric cancer

<正>Objective To explore the expression and significance of CD44 gastric cancer stem cell biomarker positive circulating tumor cells(CTC)in peripheral blood of patients with gastric cancer.Methods From March 2010 to December 2012,45 patients with gastric cancer and 20healthy individuals were enrolled.Peripheral blood samples of all the subjects were collected.The expression of     

小细胞肺癌血钙与外周循环肿瘤细胞的检测

目的研究不同分期小细胞肺癌患者血钙水平与外周血循环肿瘤细胞(CTC)的检测价值。方法选择我院84例小细胞肺癌患者为研究对象。根据国际肺癌的诊断标准对患者进行分期,局限期患者33例,广泛期患者51例。对所有患者血钙水平与外周血循环肿瘤细胞水平进行检测。结果患者的体内的血清钙以及游离钙在广泛期明显多于局限期,P0.05。患者的外周血循环肿瘤细胞≥1和≥5,广泛期明显多于局限期,P0.05。患者的血钙以及外周血循环肿瘤细胞水平会随病情发展而增加,P0.05各项指标和病情的发展程度呈现出正相关关系。结论综上所述,对小细胞肺癌患者体内血钙以及外周血循环肿瘤细胞水平的检测价值较高。对于患者的早期确诊,治疗以及病情的预后评价具有十分重要的意义。     

Detection of hepatoma cells in peripheral blood of HCC patients by nested RT-PCR

AIM:To offer a more simple method with a high sensitivity and specificity for detection of hepatoma cells in peripheral blood of the patients with HCC.METHODS:Improved nested RT-PCR method was used to detect the expression of AFP mRNA in nuclear cells separated from peripheral venous blood.RESULTS:AFP mRNA contained in tenhepatoma cells was detected from 2mL peripheral blood.CONCLUSION:The improved nested RT-PCR assay for AFP mRNA expressed in cancer cells in peripheral blood might be a valuable method for clinical diagnosis of HCC.     

实时荧光定量RT-PCR检测结直肠癌中外周血端粒酶逆转录酶mRNA的表达及其临床意义

目的:建立外周血端粒酶逆转录酶(hTERT)mRNA表达量检测的实时荧光定量逆转录聚合酶链反应(RT-PCR)系统,检测结直肠癌患者hTERT mRNA的表达并探讨其对结直肠癌微转移的诊断价值.方法:用实时荧光定量RT-PCR技术检测53例结直肠癌患者和21名健康人的外周血标本hTERT mRNA表达情况,分析其表达与肿瘤临床病理特征的关系并应用接受者操作特征曲线(ROC曲线)分析hTERT mRNA检测对结直肠癌的诊断价值.结果:结直肠癌组外周血hTERT mRNA表达水平显著高于正常对照组(t1=7.953,P＜0.05).其表达与肿瘤淋巴结转移、血行转移和TNM分期相关(t'/t=2.334,2.149,2.460,均P＜0.05)hTERT mRNA诊断结直肠癌的ROC曲线下面积0.91,诊断界值为Ct≤32.结论:应用实时荧光定量RT-PCR技术克服了传统PCR只能定性检测而不能定量检测的缺-点:外周血hTERT mRNA可作为诊断结直肠癌微转移的标记物.     

大肠癌患者外周血循环癌细胞检测的临床意义

目的探讨大肠癌外周血循环癌细胞检测的临床意义.方法以CK-20 mRNA为靶分子,RT-PCR技术检测3种癌细胞株(结肠腺癌细胞株SW-480、胃癌细胞株SGC-7901、食管癌细胞株TE-11)和67例大肠癌患者外周血细胞CK-20 mRNA的表达.结果三种癌细胞株中,只有结肠癌细胞表达CK-20 mRNA.RT-PCR技术能检出107个单核细胞中的一个肿瘤细胞;67例患者中,35例外周血CK-20 mRNA阳性,阳性率与大肠癌Dukse'分期明显相关:Dukes'A期25%,B期38.46%,C期55.17%,D期70.59%.53例手术治疗患者中,术前25例CK-20 mRNA阳性,术后30例阳性.25例术后早期短程化疗患者,化疗前阳性6例,化疗后仅2例阳性.结论大肠癌循环癌细胞(CK-20 mRNA)的检测可能成为判断大肠癌恶性程度、转移和复发以及疗效监测的客观指标.     

肺癌患者外周血循环肿瘤细胞检测结果分析

目的 探讨肺癌患者外周血循环肿瘤细胞检测结果与患者临床特征、治疗效果以及预后的关系,为肺癌的临床诊治和患者预后判断提供参考.方法 选取我院肿瘤科2016年7月到2017年12月收治的肺癌患者60例为研究对象,采用免疫磁珠阴性富集法检测患者外周血循环肿瘤细胞(CTC);根据CT检查结果,参照RECIST标准评定患者治疗效...     

巢式RT-PCR技术检测肺癌患者外周血及淋巴结中微量转移癌细胞

目的　建立检测肺癌患者外周血及淋巴结中微转移癌细胞的敏感分子检测方法 ,并探讨其临床应用价值。方法　采用巢式反转录聚合酶链反应 (RT- PCR)技术特异引物扩增细胞角蛋白 19(CK19) m RNA,检测76例肺癌患者外周血及淋巴结中 CK19m RNA的表达情况。结果　 76例肺癌患者中 39例外周血 CK19m RNA阳性 ,阳性率为 5 1.3% (39/ 76 ) ,2 7例肺良性病变中仅 1例阳性 ,阳性率 3.7% ,15例健康对照者均为阴性。全部肺癌患者 15 7枚淋巴结中 CK19m RNA阳性率 38.9% (6 1/ 15 7) ,常规病理方法检测淋巴结阳性率 14 .6 % (2 3/15 7) ,两者相比 ,P<0 .0 5 ;HE染色阴性的 134枚淋巴结中 ,经巢式 RT- PCR技术检测证实存在癌转移的阳性率为 2 8.3% (38/ 134) ,两者相比 ,P<0 .0 5。结论　巢式 RT- PCR技术是一种特异性和敏感性均较高的微量癌细胞转移检测方法 ,可能有助于肺癌微转移的早期诊断     

应用Taq Man荧光定量RT-PCR快速检测肠道病毒的研究

目的建立一种快速、灵敏、特异的荧光定量RT-PCR检测肠道病毒的方法。方法根据GenBank中肠道病毒5′UTR序列,应用生物软件在保守区设计与筛选特异引物和TaqMan探针,对荧光定量RT-PCR反应体系与条件进行优化,验证方法的特异性、敏感性和重复性。并通过对急性临床样本的检测,评价该方法的实际应用价值。结果该方法对肠道病毒包括脊髓灰质炎病毒Ⅰ、Ⅱ、Ⅲ型及柯萨奇病毒和埃可病毒等的检测有高度特异性,甲肝病毒、乙脑病毒、登革病毒、腺病毒、单纯疱疹病毒、诺如病毒等均呈阴性。该方法的检测下限达10-1TCID50/100μl。12份急性结膜炎病例结膜拭子标本中7份肠道病毒核酸阳性,普通RT-PCR方法5份阳性。结论TaqMan荧光定量RT-PCR方法快速、敏感、特异,适于肠道病毒的快速检测。     

实时RT-PCR法检测非小细胞肺癌淋巴微转移

目的以Lunx和血管内皮生长因子C(VEGF-C)及其受体为靶基因,实时PT-PCR法检测非小细胞肺癌(NSCLC)患者癌旁组织、手术切除区域淋巴结微转移,探讨人肺特异性X蛋白(humanlung-specificXprotein,Lunx)、VEGF-C、VEGF受体3(VEGFR-3)的敏感性和特异性。方法以肺部良性病变切除的20枚淋巴结为对照,采用SYBRGreenⅠ嵌合荧光实时RT-PCR检测NSCLC手术切除区域淋巴结60枚LunxmRNA表达并与常规病理比较;以肺部良性病变切除的组织20例为对照,检测VEGF-C、VEGFR-3在30例癌旁组织中的表达。结果30例NSCLC患者癌旁组织VEGFCR-3、VEGF-C阳性率分别为60%(18/30)、77%(23/30),分别与病理阳性率37%(11/30)比较,均有显著差异(χ2=4,P<0.05;χ2=6.72,P<0.01);60枚NSCLC患者手术切除区域淋巴结Lunx阳性率78.3%(47/60),与病理阳性率55%(33/60)比较有显著差异(χ2=7.68,P<0.01)。结论VEGFCR-3、VEGF-C、Lunx均可作为NSCLC淋巴微转移的分子标志物,与病理比较有较高的敏感性;实验采用一种新颖的荧光实时RT-PCR相对定量方法分析结果,有助于早期肺癌转移的检测。     

胃癌患者外周血微转移的细胞学检测

目的：肿瘤的浸润和转移是影响患者预后的重要因素，而肿瘤转移大多通过淋巴及血循环播散。为此该文研究建立胃癌患者外周血微转移癌细胞的细胞学检测方法并探讨其临床意义。方法：取进展期胃癌患者外周静脉血5ml，经Ficoll液密度梯度离心及免疫磁珠法（磁珠采用细胞角蛋白7和细胞角蛋白8包被）分离出单核细胞，然后对细胞涂片进行苏木精－伊红染色观察；并采用免疫荧光染色和免疫细胞化学法检测癌胚抗原、人端粒酶逆转录酶、CD34和CD45，对其中的胃癌细胞进行生物学特性分析。结果：在24例胃癌患者的外周血中有10例找到了癌细胞，阳性率为41．7％；癌细胞的检出与肿瘤原发灶的病理分级差异有显著性（P＜0．05）。结论：应用Ficoll密度梯度离心和免疫磁珠分选技术在胃癌患者的外周血中可分离检测到癌细胞,并可据此拟定治疗方案并对患者的预后作出判断。     

Characterization of smooth muscle-like cells in circulating human peripheral blood

Smooth muscle cells play an important role in human vascular diseases. Several lines of evidence demonstrate that circulating smooth muscle precursor cells contribute to intimal hyperplasia in animal models. We obtained large spindle cells expressing alpha-smooth muscle actin (alpha-SMA), denoted here as "smooth muscle-like cells" (SMLC), from human peripheral blood mononuclear cells (PBMC). SMLC derived from human PBMC proliferated readily and expressed pro-inflammatory genes during early culture. After long-term culture, SMLC could contract and express characteristic smooth muscle cell markers. We found peripheral blood mononuclear cell expressing alpha-smooth muscle actin in the circulating blood that bore CD14 and CD105. Sorted CD14/CD105 double-positive PBMC could differentiate into SMLC. The number of CD14-CD105-bearing PBMC increased significantly in patients with coronary artery disease compared to patients without coronary artery disease. These results support the novel concept that smooth muscle precursor cells exist in circulating human blood and may contribute to the pathogenesis of vascular diseases.     

Prognostic and predictive blood biomarkers in gastric cancer and the potential application of circulating tumor cells

Gastric cancer(GC), with its high incidence and mortality rates, is a highly fatal cancer that is common in East Asia particularly in China. Its recurrence and metastasis are the main causes of its poor prognosis. Circulating tumor cells(CTCs) or other blood biomarkers that are released into the circulating blood stream by tumors are thought to play a crucial role in the recurrence and metastasis of gastric cancer. Therefore, the detection of CTCs and other blood biomarkers has an important clinical significance; in fact, they can help predict the prognosis, assess the staging, monitor the therapeutic effects and determine the drug susceptibility. Recent research has identified many blood biomarkers in GC, such as various serum proteins, autoantibodies against tumor associated antigens, and cell-free DNAs. The analysis of CTCs and circulating cell-free tumor DNA(ctDNA) in the peripheral blood of patients with gastric cancer is called as liquid biopsy. These blood biomarkers provide the disease status for individuals and have clinical meaning. In this review, we focus on the recent scientific advances regarding CTCs and other blood biomarkers, and discuss their origins and clinical meaning.     

Quantitation of CD 44 in tumor cells and peripheral blood of patients with gastric cancer

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