Expression Levels of RFP in Normal and Cancer Human Tissues via Real-time RT-PCR Detection

IntroductionRet finger protein(RFP) was first identified as apart of the human recombined transforming gene, ret,and was a member of the tripartite motif(TRIM) pro-tein family. Itwas postulated thatmultimeric complexesmade of TRIM proteins defined differe…     

Alterations of HLA class I and II antigen expression in preinvasive, invasive and metastatic cervical cancers

HLA expression is altered in a large variety of human cancers. We performed immunohistochemical staining on tissues from normal, preinvasive, invasive and metastatic cervical cancer tissues using anti-HLA class I or class II antibody. In tissues from normal squamous epithelium, carcinoma in situ (CIS) and microinvasive carcinoma (MIC), the expressions of HLA-B, C heavy chains and class II heavy chain were significantly decreased as disease progressed. When the expression patterns were compared between primary and metastatic squamous cell carcinoma (SCC) lesions, statistically significant down-regulation of HLA class I and class II antigen in metastatic lesions was observed. The rates of HLA-B, C heavy chains and class II heavy chain expressions were all significantly down-regulated compared to the down-regulation rate of class I beta2-microglobulin (beta2m) in invasive squamous lesions, and the expressions of class II heavy chain in metastatic lesions was decreased further than that in primary lesions. Unlike SCC, the degree of HLA class I and class II loss was not evident as disease progressed in early stage of adenocarcinoma. In invasive adenocarcinoma lesions, only the expression of HLA-B, C heavy chains was decreased and no differences were seen in HLA-B, C heavy chain expression patterns between primary and metastatic lesions. These results suggest that alterations of HLA class I and II expressions seem to occur at a particular step in cervical cancer development and depend on tissue types: when the tumor becomes invasive and starts to metastasize.     

Expression of high-abundance proteins in sera of patients with endometrial and cervical cancers: analysis using 2-DE with silver staining and lectin detection methods

The expression of high-abundance serum proteins in newly diagnosed patients with endometrial adenocarcinoma (EACa), squamous cell cervical carcinoma (SCCa) and cervical adenocarcinoma (ACCa), relative to control female subjects, was analyzed by subjecting serum samples to 2-DE followed by image analysis of the silver-stained protein profiles. The three cohorts of cancer patients demonstrated different altered expression of serum high-abundance proteins compared to negative control women. The expression of alpha1-antitrypsin, alpha1-B glycoprotein, cleaved high-molecular-weight kininogen (light chain) and antithrombin III were consistently altered in all the patients. However, clusterin was upregulated only in the patients with EACa, while those with SCCa and ACCa were typically characterized by the upregulated expression of zinc alpha-2-glycoprotein. The aberrant expression of selective serum proteins in the various cohorts of cancer patients was validated by competitive ELISA as well as by lectin detection. Analysis by using the champedak galactose binding lectin further highlighted an unidentified protein that may be differently glycosylated in the sera of the EACa patients that were studied.     

Synchronous Fluorescence Spectroscopy for the Detection and Characterization of Cervical Cancers In Vitro

The objective of this study was to assess the diagnostic potential of synchronous fluorescence (SF) spectroscopy (SFS) technique for the detection and characterization of normal and different malignancy stages of moderately differentiated squamous cell carcinoma (MDSCC), poorly differentiated squamous cell carcinoma (PDSCC) cervical tissues. SF spectra were measured from 45 biopsies from 30 patients in vitro . Characteristic, highly resolved peaks and significant spectral differences between normal and MDSCC, PDSCC cervical tissues were obtained. Nine potential ratios were calculated and used as input variables for a discriminant analysis across different groups. The potentiality of the SFS technique was estimated by two discriminant analyses. Discriminant analysis I performed across normal and abnormal (including MDSCC and PDSCC) cervical tissues classified as 100% both original and the cross-validated grouped cases. In discriminant analysis II performed across the three groups, normal, MDSCC and PDSCC, 100% of both original and the cross-validated grouped cases were correctly classified. Using the SFS technique, one can obtain all the key biochemical markers such as tryptophan, collagen, hemoglobin, reduced form of nicotinamide adenine dinucleotide and flavin adenine dinucleotide in a single scan and hence they can be targeted as tumor markers in the detection of normal from abnormal cervical tissues.     

Identification of key classification features of early cervical squamous cell carcinoma

Despite the tremendous progress in molecular analysis of pan-cancer, little is known regarding molecular classification of cervical squamous cell carcinoma. In this study, we adopted a multi-omics approach to identify potential key classification features of cervical squamous cell carcinoma. Specifically, we analyzed mRNA, and microRNA (miRNA) expression data, as well as DNA methylation and copy number variation in cervical squamous cell carcinoma cases, using datasets obtained from The Cancer Genome Atlas (TCGA). Moreover, we identified molecules in each dimension, as well as integrated and clustered filtered classification features, and used them to distinguish different subtypes. The resulting key classification features were used to establish a classification model for cervical squamous cell carcinoma. Our results revealed two cervical squamous cell carcinoma subtypes, with significant differences across clinical survival levels, as well as 8 key classification features of cervical squamous cell carcinomas. These findings are expected to provide important references for early classification of cervical squamous cell carcinoma and identification of classification markers.     

Basic and clinical evaluation of a new assay procedure "SCC RIABEAD" for estimation of squamous cell carcinoma related antigen

We examined the efficacy of a new commercial assay procedure (SCC RIABEAD) for estimation of squamous cell carcinoma related antigen (SCC). Intraassay and interassay variance were 4.0-14.6% and 4.6-17.0% respectively. Recovery and dilution tests gave satisfactory results. The normal range was under the level of 1.63 ng/ml. The patients with uterine cervical cancer or squamous cell carcinoma of the lung showed high positive rates. The values of SCC measured by SCC RIABEAD were well-correlated to those by SCC RIAKIT. However, SCC RIABEAD showed lower values in low SCC level and higher values in high SCC level than SCC RIAKIT.     

Quantification of glucose diffusion in human lung tissues by using Fourier domain optical coherence tomography

In this study, we report permeability coefficients of 30% glucose diffusion by the optical coherence tomography signal slope (OCTSS) method in four kinds of human lung tissue in vitro: normal lung tissue, benign granulomatosis lung tissue, squamous cell carcinoma and adenocarcinoma tumor. To quantify the permeability coefficient of the agent, the monitored region was 80 μm thickness at a tissue depth of ca 230 μm from the surface. The permeability coefficients of 30% glucose from 10 independent experiments were averaged and found to be (1.35 ± 0.13) × 10(-5) cm s(-1) from the normal lung tissue, (1.78 ± 0.21) × 10(-5) cm s(-1) from the benign granulomatosis lung tissue, (2.88 ± 0.19) × 10(-5) cm s(-1) from the adenocarcinoma tumor and (3.53 ± 0.25) × 10(-5) cm s(-1) from the squamous cell carcinoma, respectively. It could be clearly seen that the permeability coefficients of 30% glucose increase ca 32%, 113% and 162% in the benign granulomatosis, adenocarcinoma tumor and squamous cell carcinoma of human lung tissue compared with that from the normal lung tissue, respectively. Therefore, we inferred from this pilot study that the OCT imaging is a feasible method to distinguish normal and cancer lung tissue.     

Phytochemical Profile and Anticancer Potential of Endophytic Microorganisms from Liverwort Species,Marchantia polymorpha L.

Liverwort endophytes could be a source of new biologically active substances, especially when these spore-forming plants are known to produce compounds that are not found in other living organisms. Despite the significant development of plant endophytes research, there are only a few studies describing liverwort endophytic microorganisms and their metabolites. In the presented study, the analysis of the volatile compounds obtained from thallose liverwort species, Marchantia polymorpha L., and its endophytes was carried out. For this purpose, non-polar extracts of plant material and symbiotic microorganisms were obtained. The extracts were analyzed using gas chromatography coupled to mass spectrometry. Compounds with the structure of diketopiperazine in the endophyte extract were identified. Liverwort volatile extract was a rich source of cuparane-, chamigrane-, acorane-, and thujopsane-type sesquiterpenoids. The cytotoxicity of ethyl acetate extracts from endophytic microorganisms was evaluated on a panel of cancer (FaDu, HeLa, and SCC-25) cell lines and normal (VERO), and revealed significant anticancer potential towards hypopharyngeal squamous cell carcinoma and cervical adenocarcinoma.     

Design,Synthesis, and In Vitro Anticancer Activities of Diethylene Glycol Tethered Isatin‐1,2,3‐triazole‐coumarin Hybrids

A series of novel diethylene glycol tethered isatin‐1,2,3‐triazole‐coumarin hybrids 9a – l were designed, synthesized, and evaluated for their in vitro anticancer activities against HepG2 (liver carcinoma), Hela (cervical cancer), A549 (lung adenocarcinoma), DU145 (prostatic cancer), SKOV3 (ovarian carcinoma), MCF‐7 (breast cancer), and drug‐resistant MCF‐7/DOX (doxorubicin‐resistant MCF‐7) human cancer cell lines. The results showed that most of the synthesized hybrids exhibited considerable in vitro activities against the tested seven cancer cell lines, and these hybrids can be acted as starting points for further investigation.     

Direct analysis of laser capture microdissected endometrial carcinoma and epithelium by matrix-assisted laser desorption/ionization mass spectrometry

Direct analysis of laser capture microdissected malignant and normal endometrial epithelium using matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (MS) was able to detect a number of proteins that are overexpressed in malignant epithelial cells. A total of 16 physiologic and malignant endometrial samples were laser capture microdissected, including four proliferative and four secretory endometria, and eight endometrioid adenocarcinomas. Two of these proteins, at 10,834 and 10,843 Da, likely correspond to calgranulin A and chaperonin 10, two proteins that had previously been identified in endometrioid adenocarcinoma in whole tissue homogenate by MS analysis. Direct analysis by MALDI-MS not only confirms that these proteins are overexpressed in endometrial carcinoma, but also localizes them to the epithelial cells, the expected cancer site.     

Delimitation of squamous cell cervical carcinoma using infrared microspectroscopic imaging

Infrared (IR) spectroscopic imaging coupled with microscopy has been used to investigate thin sections of cervix uteri encompassing normal tissue, precancerous structures, and squamous cell carcinoma. Methods for unsupervised distinction of tissue types based on IR spectroscopy were developed. One-hundred and twenty-two images of cervical tissue were recorded by an FTIR spectrometer with a 64×64 focal plane array detector. The 499,712 IR spectra obtained were grouped by an approach which used fuzzy C-means clustering followed by hierarchical cluster analysis. The resulting false color maps were correlated with the morphological characteristics of an adjacent section of hematoxylin and eosin-stained tissue. In the first step, cervical stroma, epithelium, inflammation, blood vessels, and mucus could be distinguished in IR images by analysis of the spectral fingerprint region (950–1480 cm⁻¹). In the second step, analysis in the spectral window 1420–1480 cm⁻¹ enables, for the first time, IR spectroscopic distinction between the basal layer, dysplastic lesions and squamous cell carcinoma within a particular sample. The joint application of IR microspectroscopic imaging and multivariate spectral processing combines diffraction-limited lateral optical resolution on the single cell level with highly specific and sensitive spectral classification on the molecular level. Compared with previous reports our approach constitutes a significant progress in the development of optical molecular spectroscopic techniques toward an additional diagnostic tool for the early histopathological characterization of cervical cancer.     

Mass spectrometry meets chip technology: a new proteomic tool in cancer research?

DNA chip technologies are the most exiting genomic tools, which were developed within the last few years. It is, however, evident that knowledge of the gene sequence or the quantity of gene expression is not sufficient to predict the biological nature and function of a protein. This can be particularly important in cancer research where post-translational modifications of a protein can specifically contribute to the disease. To address this problem, several proteomic tools have been developed. Currently the most widely used proteomic tool is two-dimensional protein gel electrophoresis (2-DE), which can display protein expression patterns to a high degree of resolution. As an alternative to 2-DE, a preliminary study using a new technique was employed to generate protein expression patterns from whole tissue extracts. Surface-enhanced laser desorption/ionization (SELDI) allows the retention of proteins on a solid-phase chromatographic surface (ProteinChip Array) with direct detection of retained proteins by time of flight-mass spectrometry (TOF-MS). Using this system, we analyzed eight cases of renal cell carcinoma (RCC) including normal, peripheral and central tumor tissue as well as four microdissected cases of cervical intraepithelial neoplasia (CIN) and three microdissected cases of cervix uteri carcinoma. Differentially expressed proteins were found by comparing the protein expression patterns generated using SELDI-based TOF-MS of tumor tissue with normal and neoplastic tissue, respectively. By applying this fast and powerful ProteinChip array technology it becomes possible to investigate complex changes at the protein level in cancer associated with tumor development and progression.     

Visnagin inhibits cervical cancer cells proliferation through the induction of apoptosis and modulation of PI3K/AKT/mTOR and MAPK signaling pathway

Cervical cancer, a silent killer is a second most common type of malignant tumor detected in women’s world wide. In modern medicine the usage of phytochemicals to develop drugs for treating various chronic diseases is rapidly increasing. One such phytochemical is visnagin, a furanochrome present in fruits of Ammi visnaga. We investigated the anticarcinogenic potency of visnagin against human cervical carcinoma cells. The antioxidant potency of visnagin was analyzed with FRAP assay, DPPH assay, Chemiluminscence assay and ORAC assay. The cytotoxic effect of visnagin on normal epithelial Vero cells and human cervical cancer HeLa cells were analyzed using MTT assay. The effect of visnagin on antioxidant system was examined by measuring the levels of TBARS, SOD and GSH using the colorimetric assay techniques. DCFH-DA staining, AO/EtBr staining, propidium iodide staining was performed to assess the apoptotic induction potency of visnagin against cervical cancer cells. The ability of visnagin to inhibit cancer cell migration was examined with scratch wound assay. The anticarcinogenic property of visnagin was confirmed by analyzing the gene expression of PI3K/AKT/mTOR signaling proteins and MAPK signaling proteins using qPCR analysis. Visnagin exhibited increased Trolox equivalent value in all the four antioxidant potency estimating experiments. Visnagin induced cytotoxic effect only on carcinoma cells, decreased the antioxidants and increased the generation of ROS. It also induced apoptosis and inhibited the cancer cell migration. The qPCR analysis confirms visnagin decreases the gene expression cell cycle regulating protein of both PI3K/AKT/mTOR and MAPK pathway. Overall our results authentically prove visnagin inhibits the progression of cervical cancer in vitro. Therefore it can be an ideal drug of choice which can subject to further investigation for treating cervical cancer.     

Photodynamic diagnosis in respiratory tract malignancy using an excimer dye laser system

Equipment has been developed for the early-stage diagnosis and treatment of cancer using an excimer dye laser. The excimer laser beam is obtained by exciting XeCl. A 405 nm beam tuned by DPS dye is used for tumour localization and a 630 nm beam obtained with a rhodamine B dye is used for treatment. The equipment was applied clinically on the basis of extensive experimental research. Effectiveness for cancer localization was examined in 11 cases: four were early stage (three lung cancer and one vocal cord cancer), four were stage I, two were stage III and one was stage IV. All cases were squamous cell carcinoma except for one case of adenocarcinoma. Fluorescence was recognized in all lesions and the equipment was effective for localization.     

Clinical evaluation of gallium-67 scintigraphy in comparison with autopsy findings in the elderly, with particular reference to histological findings and classification of pulmonary cancer

A correlative study of autopsy findings and retrospective review of gallium scintigrams were performed in 106 elderly patients. Of the cases studied, 57% demonstrated positive gallium study in the present series. Histological correlation was undertaken in cases of lung cancer. Among them, squamous cell carcinoma showed the highest incidence of positive results (83%), whereas adenocarcinoma was the lowest (35%). There is no apparent correlation between subtypes of histological classification of adenocarcinoma and abnormal accumulation of gallium. However, abnormal accumulation of the nuclide seems to be rather related with interstitial reactions, namely fibrotic changes, lymphocyte infiltration and vascularization.     

The importance of enhancer methylation for epigenetic regulation of tumorigenesis in squamous lung cancer

Lung squamous cell carcinoma (LUSC) is a subtype of non-small cell lung cancer (NSCLC). LUSC occurs at the bronchi, shows a squamous appearance, and often occurs in smokers. To determine the epigenetic regulatory mechanisms of tumorigenesis, we performed a genome-wide analysis of DNA methylation in tumor and adjacent normal tissues from LUSC patients. With the Infinium Methylation EPIC Array, > 850,000 CpG sites, including ~350,000 CpG sites for enhancer regions, were profiled, and the differentially methylated regions (DMRs) overlapping promoters (pDMRs) and enhancers (eDMRs) between tumor and normal tissues were identified. Dimension reduction based on DMR profiles revealed that eDMRs alone and not pDMRs alone can differentiate tumors from normal tissues with the equivalent performance of total DMRs. We observed a stronger negative correlation of LUSC-specific gene expression with methylation for enhancers than promoters. Target genes of eDMRs rather than pDMRs were found to be enriched for tumor-associated genes and pathways. Furthermore, DMR methylation associated with immune infiltration was more frequently observed among enhancers than promoters. Our results suggest that methylation of enhancer regions rather than promoters play more important roles in epigenetic regulation of tumorigenesis and immune infiltration in LUSC.Subject terms: Cancer genomics, Non-small-cell lung cancer     

Efficient Gaussian sample specific network marker discovery and drug enrichment analysis validation

Identifying stable gene markers at an individual level can help to understand the genetic mechanisms of each individual patient and accomplish personalized medicine. In this paper, we propose an efficient framework to identify sample-specific markers. Gene expression data first is transformed to a corresponding likelihood matrix to alleviate inherent noise besides adding population information to each sample. Then those significantly differential genes or gene pairs are further mapped to a STRING network for analysis by assuming that the likelihood of each gene or gene pairs in the control group follows a Gaussian distribution. The proposed method is applied to three benchmark datasets including lung adenocarcinoma, kidney renal clear cell carcinoma, and uterine corpus endometrial carcinoma. It is found that disease gene markers identified by the proposed methods outperform the previous sample-specific network (SSN) method in both subtyping and survival analysis. Furthermore, we exploit the application of the subtype markers in following drug selection. The difference of the enriched drug set may reflect some underlying mechanisms of the subtypes and shed light on selecting appropriate drugs for each cancer subtype.     

肺癌组织中微量元素的研究

用ICP等离子体发射光谱法测定了肺癌组织中Cd、Cr、Co、Mg、Mn、Ni、Pb、Sr、Ti、Se、Al、Ba、Zn、Fe、Cu、Be等16种微量元素古量。结果显示．与同侧无癌肺叶组织相比较，肺鳞癌和肺腺癌组织中Cr、Cu和Mg含量均显著升高．Cd、Mn和Zn含量均显著降低。两型肺癌组织中Cd、Fe和Ba3种元素音量之间存在明显差异．而两型肺癌的同侧无癌肺叶组织的微量元素含量之间除Cd外均无明显差异。     

食管癌组织中铜含量及其临床意义

应用日立１８０－３０型原子吸收火焰分光光度计，对３６例食管癌组织中的微量元素Ｃｕ进行了测定，并与３０例食管癌患者的癌旁正常食管组织进行了对照比较。结果表明，癌组织Ｃｕ含量显著高于癌旁组织Ｃｕ含量。