Partial Biological Characterization of Cancer Stem-like Cell Line (WJ<Subscript>2</Subscript>) of Human Glioblastoma Multiforme

To provide suitable models for human GBM cancer stem cells in vitro and in vivo, and investigate their biological characteristics, a new human GBM cancer stem-like cell line, WJ2, was established in this experiment through serial passages from adherent monolayer culture to nonadherent tumor sphere culture in turns; Its partial biological characteristics were studied through cell proliferation and tumor sphere assay; cell cycle distribution, side population, and CD133 phenotype were analyzed with FCM. The expressions of CD133, Nestin, and GFAP of cancer stem-like cells and xenograft tumor cells were detected with RT-PCR and immunohistochemistry. Biological characterization, side population, CD133 phenotype and CD133 Nestin, BCRP-1, Wnt-1 gene expression revealed the stemness of this cancer stem-like cell line. Tumorigenicity heterotransplanted in nude mice; histopathological characteristics of xenograft tumor, and expressions of CD133, Nestin, and GFAP of xenograft tumor cells indicated that xenograft tumors recapitulated the phenotype and biological characterization of human primary GBM. All findings of this experimental study suggested that WJ₂ cancer stem-like cell line could accurately mimic human GBM cancer stem cell in vitro and in vivo; it would be useful in the cellular and molecular studies as well as in testing novel therapies of CSC-based anti-cancer therapies for human GBM.     

High expression of osteoglycin decreases the metastatic capability of mouse hepatocarcinoma Hca-F cells to lymph nodes

Osteoglycin, one of the matrix molecules, belongs to the small leucine-rich proteoglycan gene family and might play important roles in cell growth and differentiation and in pathological processes such as fibrosis and cancer growth. In this study, a eukaryotic expression plasmid pIRESpuro3 osteoglycin(+) was constructed and transfected into mouse hepatocarcinoma Hca-F cells to evaluate the contribution of osteoglycin to the malignant behavior of Hca-F. It was found that Hca-F cells transfected with pIRESpuro3 osteoglycin (+) showed significantly decreased potential for both migration and invasion. Furthermore, Hca-F cells transfected with osteoglycin showed decreased metastatic potential to peripheral lymph nodes. However, proliferation potential and adhesive capacity of Hca-F cells to differentprotein substrates were not influenced by osteoglycin transfection. In summary, high expression of osteoglycin decreases the metastatic capability of Hca-F to lymph nodes.     

血清miRNAs作为食管鳞癌新的生物标志物

食管鳞癌是我国最常见的恶性肿瘤之一。由于缺乏有效的早期诊断方法,大多数食管鳞癌患者在确诊时已到中晚期并预后不良。MicroRNAs(miRNAs)是一类可通过抑制其特异性靶基因表达从而调控食管鳞癌发生发展的非编码内源性小RNA。相比于传统的生物标志物(例如mRNA和蛋白质),miRNAs更加稳定并易于筛选及精确地定量分析,从而成为理想的新一代癌症早期诊断和预后评估的生物标志物。近来的研究结果显示,食管鳞癌病人血清中的一些miRNAs表达水平的变化与病情诊断及预后的结果显著相关。文章综述了食管鳞癌病人血清中miRNAs的变化规律,讨论了检测这些miRNAs的表达水平变化作为一种新的方法应用于食管鳞癌的早期诊断和预后评估的可能性。值得注意的是,不同的血清miRNAs的检测方法所产生的结果是不完全一致的,文章还对这些差异产生的原因进行了讨论。     

食管癌前细胞DNA、端粒酶含量及多基因表达产物的定量检测

为探讨食管癌高发区人群食管上皮癌变过程中的早期分子改变及早期癌变机理.应用流式细胞术和免疫荧光技术及碘化丙啶DNA荧光染色方法,对食管上皮癌前细胞的DNA含量、端粒酶含量和多个基因p53、p16、cyclin D1蛋白质表达进行了定量检测.检测结果发现,DNA含量在癌变形成时明显增加,异倍体率为87.9%;p53蛋白积聚发生在癌变早期,在癌细胞组的阳性率为100%（5/5）;抑癌基因p16在癌变早期有明显缺失;癌基因cyclin D1及端粒酶阳性率在癌细胞组都为100%（分别为6/6,7/7）.研究结果表明：在癌变早期,DNA含量及异倍体率增加,癌基因cyclin D1表达增高,抑癌基因p16缺失及p53蛋白积聚,端粒酶含量也明显增高,在癌形成时已有多个分子事件发生.     

高龄患者腹腔镜结直肠癌根治术围手术期对机体应激及内脏蛋白的影响

目的:研究高龄病人(75岁以上)腹腔镜与开腹结直肠癌根治术围手术期白介素6(IL-6)、IL-10和C反应蛋白(CRP)及内脏蛋白的差异.方法:将41例行结直肠癌根治术的高龄患者按患者意愿分为腹腔镜组(n=20)和开腹组(n=21),两组患者的一般资料如年龄、性别、体重指数(BMI)等有可比性,检测两组患者术前、术后的应激指标:血清IL-6、IL-10和CRP及营养蛋白:前白蛋白(prealbumin,PRE)、转铁蛋白(transferrin,TRF)、视黄醇结合蛋(retinal-binding protein,RbP)的变化.结果:两组患者CRP在术后1、2、3d均较术前明显升高(P<0.01),术后2d达到峰值,腹腔镜组术后CRP明显低于开腹组(P<0.01);两组患者血清IL-6、IL-10术后也明显升高,腹腔镜组明显低于开腹组(P<0.01),但IL-10升高持续时间短.PRE、TRF、RbP术后两组均较术前有明显下降(P<0.01),术后1 d、2 d两组各指标无显著性差异(P>0.05),术后3 d腹腔镜组4种蛋白指标均明显高于开腹组(P<0.01).结论:对高龄病人两种不同手术方式应激水平及内脏蛋白指标的研究提示,腹腔镜结直肠癌根治术较开腹手术创伤小,应激水平低,有利于机体内脏蛋白的恢复,这对微创外科的实施是一个有力支持.     

食管癌细胞NGAL基因-152～-60区段存在TPA反应元件

以往研究发现,在TPA诱导永生化食管上皮细胞癌变中中性粒细胞明胶酶相关载脂蛋白(neutrophilgelatinase-associatedlipocalin,NGAL)基因过表达,但过表达机制不明.最近研究提示,食管癌细胞NGAL启动子及其邻近区域可能存在着TPA反应元件.为了对NGAL的这一TPA反应元件进行更准确定位,采用PCR法结合嵌套缺失实验从食管癌细胞中克隆了NGAL5′侧翼区-152～ 84、-140～ 84、-78～ 84、-59～ 84、-50～ 84、-41～ 84、-37～ 84、-29～ 84和-10～ 84等片段,并定向插入pGLB、pGLP或pGLE等萤火虫荧光素酶报告基因表达载体中,构建了pGLB-152、pGLP-152、pGLE-152、pGLB-140、pGLB-78、pGLB-59、pGLB-50、pGLB-41、pGLB-37、pGLB-29和pGLB-10等系列报告基因表达载体.将上述报告基因表达载体分别同pRL-TK共转染食管癌细胞EC109,并用TPA刺激,检测TPA刺激转染EC109的相对荧光素酶活力,综合判定NGAL-152～ 84区不同长度片段的TPA反应性,对NGAL启动子区的TPA反应元件给予进一步分段定位.结果表明,NGAL启动子区的TPA反应元件位于-152～-60区段,而且应答TPA刺激的反应能力很强.生物信息学分析结果显示,NGAL启动子区所存在的TPA反应元件很可能是一种新结构类型.研究说明,NGAL在DNA序列上有应答TPA刺激的结构基础,将有助于深入到分子水平揭示TPA诱导永生化食管上皮细胞癌变中NGAL过表达机制,也有助于进一步认识TPA信号细胞内传递途径网络在肿瘤发生发展中的作用.     

Peritoneal fluid cytology in serous borderline tumours of the ovary

stewart c. j. r. and kennedy j. h. (1998) Cytopathology 9, 38–45
Peritoneal fluid cytology in serous borderline tumours of the ovary
Peritoneal fluid cytology findings in three patients with serous borderline tumours of the ovary and peritoneal serous implants are presented. The specimens were characterized by papillary groups, acinar clusters and single neoplastic cells exhibiting cytoplasmic vacuolation and nuclear atypia of variable degree. The cytological appearances were initially considered consistent with ovarian adenocarcinoma in all cases. Histological correlation is required to avoid this diagnostic pitfall.     

PAX5 gene as a novel methylation marker that predicts both clinical outcome and cisplatin sensitivity in esophageal squamous cell carcinoma

Therapeutic strategies for esophageal cancer largely depend on histopathological assessment. To select appropriate treatments of individual patients, we examined the background molecular characteristics of tumor malignancy and sensitivity to multidisciplinary therapy. Seventy-eight surgically-resected esophageal squamous cell carcinoma (ESCC) cases during 2001–2013 were examined. PAX5, a novel gene methylation marker in ESCC, was evaluated in the specimens, as methylation of this gene was identified as an extremely tumor-specific event in squamous cell carcinogenesis of head and neck. PAX5 methylation status was evaluated by quantitative MSP (QMSP) assays. Mean QMSP value was 15.7 (0–136.3) in ESCCs and 0.3 (0–8.6) in adjacent normal tissues (P < 0.001). The 78 cases were divided into high QMSP value (high QMSP, n = 26) and low QMSP value (low QMSP, n = 52). High QMSP cases were significantly associated with downregulated PAX5 expression (P = 0.040), and showed significantly poor recurrence-free survival [Hazard Ratio (HR) = 2.84; P = 0.005; 95% Confidence Interval (CI): 1.39–5.81] and overall survival (HR = 3.23; P = 0.002; 95%CI: 1.52–7.01) in multivariable analyses with histopathological factors. PAX5-knockdown cells exhibited significantly increased cell proliferation and cisplatin resistance. PAX5 gene methylation can predict poor survival outcomes and cisplatin sensitivity in ESCCs and could be a useful diagnostic tool for cancer therapy selection.     

An examination of the marsh microhabitats of Lymnaea tomentosa and L. columella (Mollusca: Gastropoda) by path analysis

Some physical and biological characteristics of marsh microhabitats of the pulmonate snails Lymnaea tamentosa (Pfeiffer) and L. columella (Say), intermediate hosts of Fasciola hepatica in New Zealand, are compared by path analysis. The most significant variable suggested by the various path coefficients is the relative importance of flocculence of mud in the substratum. L. tomentosa was more likely to be found in or on flocculent mud, whereas L. columella appeared to prefer firm mud. Submersion of the substratum was of more direct importance to L. columella than to L. tomentosa; the effect of water on L. tomentosa was mainly indirect, since wetter microhabitats were more likely to contain flocculent mud. Results of an experiment simulating pugging of the muddy substratum by cattle suggest that both snail species are likely to be more numerous in habitats heavily pugged by cattle.