共查询到20条相似文献,搜索用时 62 毫秒
1.
Xiaowen Huang Li Li Qin Tu Jianchun Wang Wenming Liu Xueqin Wang Li Ren Jinyi Wang 《Microfluidics and nanofluidics》2011,10(6):1333-1341
Ethanol consumption is associated with the risk of breast cancer progression; however, the mechanism of relationship has not
yet been fully explained. Research on breast cancer cell migration after ethanol stimulation may give hope for a better understanding
of the disease and oncotherapy. Conventional cell migration assays such as Boyden chamber and wound-healing assays are easy
to conduct for this purpose; however, these assays have inherent limitations. In this study, we quantified the effect of ethanol
on MCF-7 hunam breast cancer cells using a microfluidics-based wound-healing assay. Wounds were prepared by partially digesting
a confluent cell sheet using parallel laminar flows in the presence of protease trypsin. The cells at the leading edge of
the wound remained intact. Cell image analysis indicates that all the cells cultured in the microdevice took on a good morphology
and monolayer growth status. Cell viability assay demonstrates that cell viability decreased with the increase in ethanol
concentration and treatment time. For 0, 22, 43, and 65 mmol/l of ethanol, cell viability after being cultured for 24 h was
100%, 99.6%, 99.4%, and 98.4%, respectively. Studying MCF-7 human breast cancer cell migration when treated with different
ethanol concentrations revealed that the cell migration distance is directly proportional with ethanol concentration. After
being cultured for 24 h at 37°C and 5% CO2, the maximal cell migration distance was 231, 283, and 332 μm for 22, 43, and 65 mmol/l ethanol, respectively; all results
were higher than the blank test (i.e., ethanol-free test, 218 μm). These findings will be beneficial in developing microfluidic
device applications for future research on breast tumor therapy in a biomimetic microenvironment and for developing new methods
for breast cancer therapy. 相似文献
2.
Fan ZhangAuthor Vitae Jinghua TianAuthor VitaeLi WangAuthor Vitae Pingang HeAuthor VitaeYong ChenAuthor Vitae 《Sensors and actuators. B, Chemical》2011,156(1):416-422
The electrochemical monitoring of glucose consumption is relevant for cell biology studies because of its wide detection range, high sensitivity and easy implementation. Whereas the glucose consumption and cell growth rate can be tightly correlated, they should also be cell population density dependent. In this work, we fabricated high sensitive enzyme electrodes for accurate monitoring of glucose consumption of cells in different growth stages. The performance of the fabricated device was firstly evaluated by cyclic voltammetry (CV) with p-benzoquinone (PBQ) as redox mediator, showing a linear response over a wide detection range (0.3-60 mM), a high sensitivity (1.61 ± 0.10 μA mM−1 mm−2 (n = 5)) and a low detection limit (80 μM). Then, daily glucose consumptions of NIH 3T3 cells in 24-well plates were determined for a period of 7 days. The results could be compared to the cell population growth curve, showing a close correlation but different behavior. We found that the increase of the glucose consumption took place prior the cell number increase but the glucose consumption per cell decreases linearly in the exponential growth stage of cells. 相似文献
3.
Murase R Abe Y Takeuchi T Nabeta M Imai Y Kamei Y Kagawa-Miki L Ueda N Sumida T Hamakawa H Kito K 《Proteomics. Clinical applications》2008,2(4):517-527
The purpose of this study is to establish a tumor marker that can be applied for the early detection and follow-up of oral cancer patients. Employing the proteomic approach using MALDI TOF-MS, 2-DE, patient's sera and culturing cell lines, the serum autoantibodies (autoAbs) were screened and the serum levels were estimated by ELISA. Targeting the tumor cell invasion into the surrounding stromal tissues, MRC-5 human fibroblasts were employed as the target cells and a mitochondrial membrane protein, sideroflexin 3 (SFXN3), was identified. The serum anti-SFXN3-autoAb levels elevated in patients with the oral squamous cell carcinoma significantly: with 77% sensitivity and 89% specificity against control samples. The serum anti-SFXN3-autoAb levels were mildly correlated with the primary tumor sizes, however, the levels were slightly highly elevated in T1 early cancer. An immunohistochemical analysis revealed that the SFXN3 protein is expressed in the stromal fibroblasts between the caner nests and also in the basal layer of the squamous epithelium. Changes in the serum anti-SFXN3-autoAb levels after therapy correlated with the clinical tumor burden. These findings demonstrated that the serum anti-SFXN3-autoAb is worthy of clinical evaluation as a potentially of the novel tumor maker for the early detection of oral squamous cell carcinoma. 相似文献
4.
Alvarez I Carrascal M Canals F Muixí L Abián J Jaraquemada D 《Proteomics. Clinical applications》2007,1(3):286-298
HLA class I molecules present peptides on the cell surface to CD8(+) T cells. The repertoire of peptides that associate to class I molecules represents the cellular proteome. Therefore, cells expressing different proteomes could generate different class I-associated peptide repertoires. A large number of peptides have been sequenced from HLA class I alleles, mostly from lymphoid cells. On the other hand, T cell immunotherapy is a goal in the fight against cancer, but the identification of T cell epitopes is a laborious task. Proteomic techniques allow the definition of putative T cell epitopes by the identification of HLA natural ligands in tumor cells. In this study, we have compared the HLA class I-associated peptide repertoire from the hepatocellular carcinoma (HCC) cell line SK-Hep-1 with that previously described from lymphoid cells. The analysis of the peptide pool confirmed that, as expected, the peptides from SK-Hep-1 derive from proteins localized in the same compartments as in lymphoid cells. Within this pool, we have identified 12 HLA class I peptides derived from HCC-related proteins. This confirms that tumor cell lines could be a good source of tumor associated antigens to be used, together with MS, to define putative epitopes for cytotoxic T cells from cancer patients. 相似文献
5.
A dielectrophoresis-based microdevice coated with nanostructured TiO2 for separation of particles and cells 总被引:1,自引:1,他引:0
E. Morganti C. Collini R. Cunaccia A. Gianfelice L. Odorizzi A. Adami L. Lorenzelli E. Jacchetti A. Podest�� C. Lenardi P. Milani 《Microfluidics and nanofluidics》2011,10(6):1211-1221
In this study, we present a microdevice coated with titanium dioxide for cells and particles separation and handling. The microsystem consists of a pair of planar interdigitated gold micro-electrode arrays on a quartz substrate able to generate a traveling electric completed with a microfabricated three-dimensional glass structure for cell confinement. Dielectrophoretic forces were exploited for both vertical and lateral cell motions. In order to provide a biocompatible passivation layer to the electrodes a highly biocompatible nanostructured titanium dioxide film was deposited by supersonic cluster beam deposition (SCBD) on the electrode array. The dielectrophoretic effects of the chip were initially tested using polystyrene beads. To test the biocompatibility and capability of dielectrophoretic cell movement of the device, four cell lines (NIH3T3, SH-SY5Y, MDCK, and PC12) were used. Separation of beads from SH-SY5Y cells was also obtained. 相似文献
6.
Pancreatic cancer is a highly lethal disease that is difficult to diagnose at early stage and even more difficult to cure. SW1990 and PANC-1 represent the two cancer cell lines, which are both derived from pancreatic duct, but at different cell differentiation stages. In this study, we applied the iTRAQ-labeling technology and 2-D strong cation exchange/reversed phase liquid chromatography – LC-MS/MS) to profile the secreted proteins of SW1990 and PANC-1 cells in a conditioned cell culture medium. A total of 401 proteins were identified by MS/MS and protein database searching, the percentages of these proteins predicted in the categories of plasma membrane, intracellular and secreted proteins were 29.2, 32.7 and 38.2%, respectively. Fifty six proteins were identified with unknown functions and 19 proteins were quantified with significant level changes between the two cancer cell lines under the specific cell condition with 12 proteins being up-regulated (>1.3-fold change) in PANC-1 (e.g. FLJ31222 protein, 97 kDa protein, type IV collagenase precursor, 38 kDa protein and centaurin) and seven proteins being up-regulated in SW1990 (e.g. fibroblast growth factor receptor substrate 2, putative p150, hypothetical protein LOC 654463 and LOC 55701). The proteins with significant level changes may provide a baseline to investigate mechanisms underlying the differentiation of two cell lines and can be further screened for better protein biomarkers in pancreatic cancer. 相似文献
7.
The insulin-like growth factor-1 receptor (IGF-1R) plays a key role in proliferation, growth, differentiation, and development of several human malignancies including breast and pancreatic adenocarcinoma. IGF-1R targeted immunotherapeutic approaches are particularly attractive, as they may potentially elicit even stronger antitumor responses than traditional targeted approaches. Cancer peptide vaccines can produce immunologic responses against cancer cells by triggering helper T cell (Th) or cytotoxic T cells (CTL) in association with Major Histocompatibility Complex (MHC) class I or II molecules on the cell surface of antigen presenting cells. In our previous study, we set a technique based on molecular docking in order to find the best MHC class I and II binder peptides using GOLD. In the present work, molecular docking analyses on a library consisting of 30 peptides mimicking discontinuous epitopes from IGF-1R extracellular domain identified peptides 249 and 86, as the best MHC binder peptides to both MHC class I and II molecules. The receptors most often targeted by peptide 249 are HLA-DR4, HLA-DR3 and HLA-DR2 and those most often targeted by peptide 86 are HLA-DR4, HLA-DP2 and HLA-DR3. These findings, based on bioinformatics analyses, can be conducted in further experimental analyses in cancer therapy and vaccine design. 相似文献
8.
Gy-Yi Chao Tung-I Tsai Te-Jung Lu Hung-Chang Hsu Bo-Ying Bao Wan-Yu Wu Miao-Ting Lin Te-Ling Lu 《Expert systems with applications》2011,38(7):7963-7969
Bladder cancer is a common urologic cancer. Radiotherapy plays an increasingly important role in treatment bladder cancer due to radiotherapy preserves normal bladder function. However, the five-year survival rate after radiotherapy for bladder cancer patients is 30–50%. Some biological proteins influence the outcome of radiotherapy. One or two specific proteins may not be sufficient to predict the effect of radiotherapy, analyzing multiple oncoproteins and tumor suppressor proteins may help the prediction. At present, no effective technique has been used to predict the outcome of radiotherapy by multiple protein expression file from a very limited number of patients. The bootstrap technique provides a new approach to improve the accuracy of prediction the outcome of radiotherapy in small dataset analysis. In this study, 13 proteins in each cell line from individual patient were measured and then cell viability was determined after cells irradiated with 5, 10, 20, or 30 Gy of cobalt-60. The modeling results showed that when the number of training data increased, the learning accuracy of the prediction the outcome of radiotherapy was enhanced stably, from 55% to 85%. Using this technique to analyze the outcome of radiotherapy related to protein expression profile of individual cell line provides an example to help patients choosing radiotherapy for treatment. 相似文献
9.
Skvortsov S Sarg B Lindner H Lukas P Hilbe W Zwierzina H Skvortsova I 《Proteomics. Clinical applications》2008,2(6):908-914
The monoclonal antibody cetuximab directed against the epidermal growth factor receptor (EGFR) is an attractive agent for targeted therapy in advanced colorectal cancer (CRC), especially when combined with 5-fluorouracil (5-FU)-based chemotherapy. However, the mechanisms of cetuximab activity as chemosensitizer remain poorly understood. Using proteome-fluorescence-based technology, we found that cetuximab is able to suppress the expression of thymidylate synthase (TS), which is involved in the mechanism of 5-FU action. Caco-2, HRT-18, HT-29, WiDr and SW-480 CRC cells were found to express EGFR. SW-620 was used as EGFR-negative cell line. Only in EGFR-expressing cells cetuximab is able to inhibit TS expression. Combined treatment with cetuximab and 5-FU revealed a synergistic anti-tumor response that is closely correlated with functional activity of EGFR/mitogen-activated protein kinase (MAPK) pathway. Moreover, no correlation was seen between constitutive TS protein expression, level of cetuximab-induced TS down-regulation and response either to 5-FU alone or in combination with cetuximab. We demonstrated that only EGFR expression with high functional activity of EGFR/MAPK pathway is important for the synergistic effects between cetuximab and 5-FU in the investigated cell lines. 相似文献
10.
Ogura S Kaneko K Miyajima S Ohshima K Yamaguchi K Mochizuki T 《Proteomics. Clinical applications》2008,2(12):1620-1627
Proteins secreted from specific cancer cells have a high potential for use as tumor markers. We identified secreted proteins produced by 15 different carcinoma cell lines grown in serum-free medium using MS/MS. Proneurotensin/neuromedin N (proNT/NMN) was found in conditioned medium from four of seven small cell lung carcinoma cell lines but not from eight nonsmall cell lung carcinoma cell lines. These results indicate proNT/NMN has potential as a specific tumor marker of small cell lung carcinoma. 相似文献
11.
Jiang J Parker CE Hoadley KA Perou CM Boysen G Borchers CH 《Proteomics. Clinical applications》2007,1(12):1651-1659
The epidermal growth factor receptor (EGFR) is highly expressed in a variety of tumors, and is therefore an important biomarker for cancer diagnosis and a target for cancer therapy. We have developed a novel peptide-based immuno tandem mass spectrometry (iMALDI) diagnostic assay for highly sensitive, highly specific, and quantitative analysis of EGFR, which we have applied to the detection of the EGFR peptide in three cell lines and in a tumor biopsy sample. This assay is capable of detecting the EGFR target peptide bound to the antibody beads at attomole levels. The ability to directly obtain amino acid sequence data by MS/MS on any affinity-captured peptides provides specificity to this diagnostic technique. This avoids the problem of "false positives" which can result from the nonspecific binding that can occur with any affinity-based technique. The addition of stable-labeled versions of the target peptide (synthesized from stable-isotope coded amino acids) as internal standards allows absolute quantitation of the target protein. 相似文献
12.
13.
Natalie Kurganovs Hanzhi Wang Xiaoyong Huang Vladimir Ignatchenko Andrew Macklin Shahbaz Khan Michelle R. Downes Paul C. Boutros Stanley K. Liu Thomas Kislinger 《Proteomics. Clinical applications》2021,15(5):2100037
To model the problem of radiation resistance in prostate cancer, cell lines mimicking a clinical course of conventionally fractionated or hypofractionated radiotherapy have been generated. Proteomic analysis of radiation resistant and radiosensitive DU145 prostate cancer cells detected 4410 proteins. Over 400 proteins were differentially expressed across both radiation resistant cell lines and pathway analysis revealed enrichment in epithelial to mesenchymal transition, glycolysis and hypoxia. From the radiation resistant protein candidates, the cell surface protein CD44 was identified in the glycolysis and epithelial to mesenchymal transition pathways and may serve as a potential therapeutic target. 相似文献
14.
A high-throughput label-free cell assay for modulating cell-cell communication is demonstrated with the Epic? system, a resonant waveguide grating sensor platform. Natural killer (NK) cells are known to be able to recognize abnormal cells (e.g., cancer cells and cells presenting intercellular adhesion molecule 1 [ICAM1] through cell surface receptors) and kill them. In this study, the effect of effecter cells NK92MI on two kinds of target cells, cervical cancer cells (HeLa) and Chinese hamster ovarian cells overexpressing ICAM1 (CHO-ICAM1), was examined. Living target cells' response to NK92MI cells was monitored in real time and measured as wavelength shift in picometers. The authors showed that the detectability of target cell response is affected by multiple factors: the ratio of effecter cells to target cells (E/T), the interaction time of the two types of cells, and the target cell type. For example, with the effecter cells NK92MI and the same incubation time of 16 h, a minimal E/T ratio of 1 is required to detect HeLa cell response, whereas an E/T of 0.5 is sufficient to detect CHO-ICAM1 cell response. The authors confirmed that NK92MI cell-mediated target cell cytotoxicity results in negative optical signals and is associated with apoptosis mainly through caspase pathways. Distinct optical signals could be generated with the pretreatment of the target cells with various known pharmaceutical reagents, making the assay useful for discovering new chemicals that may affect cell-cell communications. 相似文献
15.
Cheng-Hsin Chuang Yuan-Chu Yu Da-Huei Lee Ting-Feng Wu Cheng-Ho Chen Shih-Min Chen Hsun-Pei Wu Yao-Wei Huang 《Microfluidics and nanofluidics》2014,16(5):869-877
This work presents a module-level impedance measurement system integrated with a disposable immunosensor for the immunoassay of bladder cancer cell lysate (T24) to a specific antibody (galectin-1). The immunosensor consisted of a flexible printed circuit patterned with an interdigital microelectrode array which immobilized polyaniline-modified nanoprobes on an electrode surface by dielectrophoresis. A quantitative sampling of cell lysate without a pump was made by using paper as the cell lysate carrier and sweeping a moistened paper over the sensing area of interdigital microelectrode array for sampling. In this study, the impedance measurement results of the module-level system were compared with those measured by the precision LCR meter, in which the error is <2 %. Additionally, the normalized impedance variation in immunosensing linearly increased with the cell lysate concentration. With a sensitivity based on a normalized impedance variation of 124.4 % per mg/ml, the immunosensor can rapidly detect the lowest concentration of cell lysate for 0.0626 mg/ml in 10 min. Therefore, this work has demonstrated the accuracy of the module-level immunosensor as well as the reliability of impedance-based sensing for bladder cancer cell lysate. The proposed disposable sensor and portable impedance system module are highly promising for use in point-of-care diagnostics. 相似文献
16.
van Oostrum J Calonder C Rechsteiner D Ehrat M Mestan J Fabbro D Voshol H 《Proteomics. Clinical applications》2009,3(4):412-422
Controlling aberrant protein kinase activity is a promising strategy for a variety of diseases, particularly cancer. Hence, the development of kinase inhibitors is currently a focal point for pharmaceutical research. In this study we utilize a chip-based reverse phase protein array (RPA) platform for profiling of kinase inhibitors in cell-based assays. In combination with the planar wave-guide technology the assay system has an absolute LOD down to the low zeptomole range. A431 cell lysates were analyzed for the activation state of key effectors in the epidermal growth factor (EGF) and insulin signaling pathways to validate this model for compound screening. A microtiter-plate format for growing, treating, and lysing cells was shown to be suitable for this approach, establishing the value of the technology as a screening tool for characterization of large numbers of kinase inhibitors against a wide variety of cellular signaling pathways. Moreover, the reverse array format allows rapid development of site-specific phosphorylation assays, since in contrast to ELISA type systems only a single antigen-specific antibody is required. 相似文献
17.
Dihazi H Müller C Asif AR Flad T Elmaouhoub A Müller GA 《Proteomics. Clinical applications》2007,1(2):200-214
For most cancers, the patient's prognosis improves dramatically if the disease is detected at an early stage. Although advancements in imaging technology have improved early detection, many cancers remain undetected until it is too late for curative intervention. We have established, for the first time, expression difference mapping analysis of whole cell proteins from renal cell carcinoma (RCC) cell lines using ProteinChip technology. A total of 20 different RCC cell lines were cultured in vitro directly on ProteinChip arrays for 24 h. Direct MS analysis of proteins from the attached cells showed identical protein profiles by all analysed RCC lines. Comparative on‐chip analysis of isolated malignant cells from native tumour specimens revealed protein patterns highly similar to those from the continuous RCC lines. However, cultured primary cortex cells showed specific protein differences. Differential protein profiling of isolated cytosolic and enriched membrane fractions from the RCC lines revealed that the protein pattern of the membrane proteins included or were identical to those of the entire cells. Proteomics analysis of the chip‐binding membrane fractions allowed the identification of three forms of galectin‐1 as potential RCC marker. ProteinChip analysis with a bound‐specific antibody certified that galectin‐1 could be an RCC marker. Immunostaining methods confirmed the overexpression of galectin‐1 in renal carcinoma in comparison to healthy tissue. 相似文献
18.
Han Wang Zhongzheng Liu Dong M. Shin Zhuo G. Chen Younghak Cho Yong-Joe Kim Arum Han 《Microfluidics and nanofluidics》2018,22(6):68
Analyzing cancer cell compressibility with single-cell resolution is of high interest in understanding cancer metastasis as well as in other cell biology applications. Here, a multi-frequency acoustophoretic cell alignment technique to precisely control cell positions in 3D space was developed, and combined with our recently developed numerical acoustophoresis model, which allowed extracting the compressibility of cells with high accuracy. This technology was applied to measure the compressibility of different head and neck cancer (HNC) cell lines that have different metastatic potential. This method can be a simple, non-contact, accurate, and low-cost solution for studying cell biomechanics and utilizing such biomechanical properties in evaluating the metastatic potential of cancer cells. 相似文献
19.
Li J Abraham S Cheng L Witzmann FA Koch M Xie J Rahman M Mohammed SI 《Proteomics. Clinical applications》2008,2(1):78-89
Identification of reliable non-invasive markers for the detection of invasive phenotype of urothelial carcinoma is needed. This study characterizes and compares protein expression profiles of adjacent non-neoplastic urothelium and invasive urothelial carcinoma to identify biomarkers for early detection of de novo bladder cancer. Differences in protein expression between adjacent non-neoplastic and high-grade, stage T4, grade 3 invasive urothelial carcinoma tissues were investigated using 2-DE, MALDI-TOF-MS, and data processing. Ingenuity Pathway Analysis (IPA) was applied to examine the biological mechanisms represented by the altered proteins. The 2-DE of the adjacent non-neoplastic urothelium and invasive urothelial carcinoma showed reproducibly similar proteomic mapping for each group distinguishing adjacent non-neoplastic urothelium from invasive urothelial carcinoma. Twenty-one proteins were altered in expression and one of these proteins, Choroideremia-like protein (CHML) was significantly overexpressed (p<0.005) and therefore was analyzed further using IHC and Western blot. Urothelial carcinoma presented an elevated expression of CHML but not adjacent non-neoplastic or normal bladder tissues. IPA revealed the involvement of CHML in cell morphology, cellular assembly, and organization. Further investigation is warranted to elucidate the biological significance of CHML and to validate its role as a biomarker for early detection of invasive urothelial carcinoma de novo. 相似文献
20.
Cell count is an important task for obtaining biological and medical information. In this paper, a novel cell count method is presented for improving the efficiency of the procedure as well as reducing microbial contamination compared to the conventional cell count method using a hemocytometer. The proposed method involves a lattice array consisting of a 50 μm × 50 μm square with lines of 2-μm width and 1.4-μm depth on a surface indented from a culture dish bottom. This configuration enables observation of cells at the same focus as the lattice. Therefore, an instant cell count during incubation is possible without the tedious, error-prone preparation steps such as harvesting and loading required in conventional methods. In addition, cells can be preserved with minimal contact with the external environment. These advantages become magnified with a periodic long-term cell count. A polystyrene culture dish, 35 mm in diameter, was fabricated by injection molding using a nickel mold, wherein indents of 3 mm × 3 mm in area and 1 mm in height were electroplated based on microfabrication technology. For easy separation from the nickel mold, the four sides of the indents in the mold are inclined at 54.74° via anisotropic silicon etching. The usefulness of the suggested method was verified using adhering HeLa (cervical carcinoma cell) cells and floating Jurkat cells. Both were placed in culture dishes and cultivated for 3 days in a carbon dioxide incubator (5% CO2, 95% air) at 37 °C, and then successfully observed through the divided lattice using an inverted microscope. The dish was also assessed with a hemocytometer by counting HEK 293T (human embryonic kidney) cells and yeast cells. 相似文献