早期宫颈癌患者术前外周血CK19 mRNA的检测及其临床意义

目的研究早期宫颈癌患者外周血CK19 mRNA检测及其临床意义.方法昆明医科大学第三附属医院妇科30例早期(ⅠA2~ⅡA)宫颈癌患者为实验组,15例子宫肌瘤患者和15例健康志愿者为阴性对照组,15例晚期(ⅡB~ⅢB)宫颈癌患者为阳性对照组.收集外周血样本,qRT-PCR技术检测外周血CK19表达水平的变化,分析其与临床病理参数、预后的相关性.结果实验组外周血中CK19表达水平与阴性对照组、阳性对照组比较,差异有统计学意义(P<0.01);30例宫颈癌患者手术前外周血CK19 mRNA阳性检出率为30%(9/30),与淋巴结转移情况、肌层浸润、术后病理高危因素密切相关(P<0.05),而与患者临床病理因素如年龄、临床分期(FIGO)、肿瘤细胞分化程度、肿瘤直径、病理类型之间均无明显相关性(P>0.05).结论宫颈癌根治术前外周血CK19 mRNA表达水平可作为诊断早期宫颈癌循环肿瘤细胞的标志物,能够弥补影像学、肿瘤标志物等检查不足,可能作为预测复发转移、判断预后的辅助指标,为术后辅助治疗方案选择、判定预后提供依据.     

KAP1在食管鳞癌中的表达及意义

目的 研究KRAB相关蛋白1(KAP1)在食管鳞癌(ESCC)中的表达及意义。方法 选取手术切除的原发性ESCC136例、高级别上皮瘤变(HGIN)35例、低级别上皮瘤变(LGIN)29例和距肿瘤边缘5cm以上的正常食管黏膜组织(NEE)37例，采用免疫组化SP法对以上各组织中KAP1蛋白的表达情况进行检测，分析其与ESCC肿瘤发生发展及患者预后的相关性；采用实时荧光定量聚合酶链反应(qRT-PCR)对20例匹配的ESCC和NEE组织中KAP1基因的表达进行检测。结果 ①KAP1蛋白的表达从NEE (24.3%)、LGIN(31.0%)、HGIN(45.7%)及ESCC(55.9%)呈逐渐增高趋势，差异均具有统计学意义(P=0.002)；②KAP1的异常表达与pTNM分期、浸润深度及淋巴结转移相关(P<0.05)；③qRT-PCR结果显示，与配对的NEE组织相比，ESCC组织中KAP1的相对表达量明显升高(P<0.01)；④136例ESCC患者的5年总体生存率为13.2%。ESCC患者的5年生存率与组织学分级、pTNM、浸润深度、淋巴结转移等具有相关性(P<0.05)，KAP1的表达与ESCC患者的预后不相关。结论 KAP1的过表达在ESCC的发生、发展和转移中起重要作用，可作为理想的肿瘤标志物辅助ESCC 诊断、指导临床治疗。     

MicroRNA检测方法的研究进展

MicroRNAs（miRNAs）是一类内源性的小分子单链非编码RNA,通过调节基因的表达在许多生命活动中起重要作用。在一些疾病（如癌症和自身免疫性疾病）发生时,miRNAs的表达谱可能发生改变,所以在药物的安全性评价中,其有望成为诊断或预后生物标志物。因此,准确测定miRNA的表达对于其应用十分重要。对传统的RNA印记（Northernblotting）、微阵列（microarray）和实时定量PCR（qRT-PCR）以及一些新的miRNAs检测方法（如基于纳米材料的miRNAs检测、核酸扩增等技术）进行概述,并阐述了这些方法的优缺点。     

Chromosome Abnormalities and Absolute Telomere Lengths of Leukocytes from Silk Weavers with Emphasis on Potential Genotoxicity and Mutagenicity of Silk Dyes

Objectives: This study is aimed to assess the possible genotoxicity and mutagenicity of silk dyes on silk weavers.Methods: Peripheral blood leukocytes were obtained from 24 silk weavers and 24 age- and sex-matched controls innortheastern Thailand. After mitogen stimulation in culture, chromosome abnormalities were examined using Giemsabanding and the absolute telomere length (aTL) was measured with SYBR green qRT-PCR. To confirm genotoxic andmutagenic effects of silk dyes, leukocytes from one each of healthy male and female volunteers were cultured withvarious concentrations of 3 dark red silk dyes under the presence of mitogen. Chromosome abnormalities and thetelomere length were determined as above. Results: The proportion of normal metaphase in the silk weaving workerswas significantly lower than that in controls. The frequency of chromosome aberrations was higher in the silk weaversthan in control group. Polyploidy was detected only in the silk weavers. The aTL was significantly shorter in the silkweavers than in control group (p < 0.05). When leukocytes from normal volunteers were stimulated with mitogen underthe presence of various concentrations of 3 silk dyes, suppressed the mitotic index (MI) and normal metaphase, whereasthe proportion of prophase and the incomplete chromosome forming increased significantly. All dyes induced polyploidy.Dye #CA5 induced structural changes in male leukocytes, whereas #30 induced the changes in female leukocytes.The #CA5 increased aTL of normal leukocytes in a dose-dependent manner. Conclusions: All dyes, especially #CA5,have high genotoxicity and mutagenicity to induce chromosome aberrations and telomeric instability. Taken all thoseresults together, regular health checking of silk weavers who have been exposed to those dyes is critically necessaryto prevent various chemical-induced carcinogenesis.