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Shi Z.,Raybiotech, Inc. | Shi Z.,South China Biochip Research Center | Yang W.-M.,Wuxi Maternal and Child Health Hospital | Chen L.-P.,Guangzhou University | And 8 more authors.
Breast Cancer Research and Treatment | Year: 2012

Drug resistance remains a major hurdle to successful cancer treatment. Many mechanisms such as overexpression of multidrug-resistance related proteins, increased drug metabolism, decreased apoptosis, and impairment of signal transduction pathway can contribute multidrug resistance (MDR). Recent studies strongly suggest a close link between cytokines and drug resistance. To identify new targets involved in drug resistance, we established a multidrug-resistant human breast cancer cell line MCF-7/R and examined the cytokine profile using cytokine antibody array technology. Among 120 cytokines/ chemokines screened, IL-6, IL-8, and 13 other proteins were found to be markedly increased in drug-resistant MCF-7/R cell line as compared to sensitive MCF-7/S cell line, while 7 proteins were specifically reduced in drugresistant MCF-7/R cells. Neutralizing antibodies against IL-6 and IL-8 partially reversed the drug resistance of MCF-7/R to paclitaxel and doxorubicin, while a neutralizing antibody against MCP-1 had no significant effect. Inhibition of endogenous IL-6 or IL-8 by siRNA technology significantly enhanced drug sensitivity of MCF-7/R cells. Furthermore, overexpression of IL-6 or IL-8 expression by transfection increased the ADM resistance in MCF-7/S cells. Our data suggest that increased expression levels of IL-6 and IL-8 may contribute to MDR in human breast cancer cells. © Springer Science+Business Media, LLC. 2012.


Burgess R.,Raybiotech, Inc. | Burgess R.,University of Texas at Dallas | Huang R.-P.,Raybiotech, Inc. | Huang R.-P.,South China Biochip Research Center
Advances in Clinical Chemistry | Year: 2015

Cancer is a complex disease involving hundreds of pathways and numerous levels of disease progression. In addition, there is a growing body of evidence that the origins and growth rates of specific types of cancer may involve "cancer stem cells," which are defined as "cells within a tumor that possess the capacity to self-renew and to cause the development of heterogeneous lineages of cancer cells that comprise the tumor. 1 1American Association of Cancer Research consensus panel definition." Many types of cancer are now thought to harbor cancer stem cells. These cells themselves are thought to be unique in comparison to other cells types present within the tumor and to exhibit characteristics that allow for the promotion of tumorigenesis and in some cases metastasis. In addition, it is speculated that each type of cancer stem cell exhibits a unique set of molecular and biochemical markers. These markers, alone or in combination, may act as a signature for defining not only the type of cancer but also the progressive state. These biomarkers may also double as signaling entities which act autonomously or upon neighboring cancer stem cells or other cells within the local microenvironment to promote tumorigenesis. This review describes the heterogeneic properties of cancer stem cells and outlines the identification and application of biomarkers and signaling molecules defining these cells as they relate to different forms of cancer. Other examples of biomarkers and signaling molecules expressed by neighboring cells in the local tumor microenvironment are also discussed. In addition, biochemical signatures for cancer stem cell autocrine/paracrine signaling, local site recruitment, tumorigenic potential, and conversion to a stem-like phenotype are described. © 2016 Elsevier Inc.


Jones V.S.,Raybiotech, Inc. | Huang R.-Y.,Raybiotech, Inc. | Chen L.-P.,Guangzhou University | Chen Z.-S.,St. John's University | And 3 more authors.
Biochimica et Biophysica Acta - Reviews on Cancer | Year: 2016

The development of oncoprotein-targeted anticancer drugs is an invaluable weapon in the war against cancer. However, cancers do not give up without a fight. They may develop multiple mechanisms of drug resistance, including apoptosis inhibition, drug expulsion, and increased proliferation that reduce the effectiveness of the drug. The collective work of researchers has highlighted the role of cytokines in the mechanisms of cancer drug resistance, as well as in cancer cell progression. Furthermore, recent studies have described how specific cytokines secreted by cancer stromal cells confer resistance to chemotherapeutic treatments. In order to gain a better understanding of mechanism of cancer drug resistance and a prediction of treatment outcome, it is imperative that correlations are established between global cytokine profiles and cancer drug resistance. Here we discuss the recent discoveries in this field of research and discuss their implications for the future development of effective anti-cancer medicines. © 2016.


Jones V.S.,Raybiotech, Inc. | Wu J.,Sun Yat Sen University | Zhu S.-W.,Raybiotech, Inc. | Huang R.-P.,Raybiotech, Inc. | Huang R.-P.,South China Biochip Research Center
Expert Reviews in Molecular Medicine | Year: 2016

Eye-derived fluids, including tears, aqueous humour and vitreous humour often contain molecular signatures of ocular disease states. These signatures can be composed of cytokines, chemokines, growth factors, proteases and soluble receptors. However, the small quantities (<10 µl) of these fluids severely limit the detection of these proteins by traditional enzyme-linked immunosorbent assay or Western blot. To maximise the amount of information generated from the analysis of these specimens, many researchers have employed multiplex immunoassay technologies for profiling the expression or modification of multiple proteins from minute sample volumes. Copyright © Cambridge University Press 2016


Browne A.S.,Emory University | Yu J.,Emory University | Yu J.,Medical Center Boulevard | Huang R.-P.,Emory University | And 7 more authors.
Fertility and Sterility | Year: 2012

Objective: To evaluate neurotrophin (NT) expression in the endometrium of women with and without endometriosis. Design: Prospective, cross-sectional, translational study. Setting: Academic hospital. Patient(s): Thirty-three reproductive-age women undergoing laparoscopy for infertility, pelvic pain, intramural fibroids, or tubal ligation. Intervention(s): Endometrial biopsies, protein microarrays, reverse transcriptase-polymerase chain reaction, ELISAs, and Western blotting. Main Outcome Measure(s): Neurotrophin proteins and mRNAs in eutopic endometrial biopsies. Result(s): Among seven neurotrophic proteins detected on the antibody microarrays, reverse transcriptase-polymerase chain reaction analysis confirmed nerve growth factor, NT-4/5, and brain-derived neurotrophic factor mRNAs in endometrial tissue. Quantitative ELISAs revealed that NT-4/5 (806 ± 701 vs. 256 ± 190 pg/100 mg protein) and brain-derived neurotrophic factor (121 ± 97 vs. 14 ± 11 ng/100 mg protein) concentrations were significantly higher in women with endometriosis. Nerve growth factor (100 ± 74 vs. 93 ± 83 pg/100 mg protein) levels did not differ between cases and controls. Conclusion(s): Neurotrophins are synthesized in situ within the endometrium. NT-4/5 and brain-derived neurotrophic factor proteins were more concentrated in biopsies from endometriosis cases than controls, whereas nerve growth factor levels were similar. We hypothesize that the local production of NTs induces sensory innervation of endometrium of women with endometriosis. These NTs represent novel targets for the diagnosis and treatment of endometriosis. © 2012 by American Society for Reproductive Medicine.


Zhou Q.,Sun Yat Sen University | Mao Y.-Q.,Raybiotech, Inc. | Jiang W.-D.,Raybiotech, Inc. | Chen Y.-R.,Raybiotech, Inc. | And 8 more authors.
PLoS ONE | Year: 2012

Purpose: Our objective was to develop a system to simultaneously and quantitatively measure the expression levels of the insulin-like growth factor (IGF) family proteins in numerous samples and to apply this approach to profile the IGF family proteins levels in cancer and adjacent tissues from patients with hepatocellular carcinoma (HCC). Experimental Design: Antibodies against ten IGF family proteins (IGF-1, IGF-1R, IGF-2, IGF-2R, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-6, and Insulin) were immobilized on the surface of a glass slide in an array format to create an IGF signaling antibody array. Tissue lysates prepared from patient's liver cancer tissues and adjacent tissues were then applied to the arrays. The proteins captured by antibodies on the arrays were then incubated with a cocktail of biotinylated detection antibodies and visualized with a fluorescence detection system. By comparison with standard protein amount, the exact protein concentrations in the samples can be determined. The expression levels of the ten IGF family proteins in 25 pairs of HCC and adjacent tissues were quantitatively measured using this novel antibody array technology. The differential expression levels between cancer tissues and adjacent tissues were statistically analyzed. Results: A novel IGF signaling antibody array was developed which allows the researcher to simultaneously detect ten proteins involved in IGF signal pathway with high sensitivity and specificity. Using this approach, we found that the levels of IGF-2R and IGFBP-2 in HCC tissues were higher than those in adjacent tissues. Conclusion: Our IGF signaling antibody array which can detect the expression of ten IGF family members with high sensitivity and specificity will undoubtedly prove a powerful tool for drug and biomarker discovery. © 2012 Zhou et al.


Huang R.-P.,Raybiotech, Inc. | Huang R.-P.,South China Biochip Research Center | Burkholder B.,Raybiotech, Inc. | Jones V.S.,Raybiotech, Inc. | And 6 more authors.
Current Proteomics | Year: 2012

Many normal physiological and disease-related pathophysiological processes are regulated by the interactions of complex signaling networks of pro- and anti-inflammatory markers, growth factors, soluble receptors and extracellular matrix proteins, as well as other cell-cell signaling proteins, which we define collectively as cytokines. Because multiple cell-cell signaling factors may be involved in a single biological process, detection of expression of multiple cytokines is essential to unraveling the mechanisms and effects of many disease processes. Cytokine antibody arrays have been developed to meet this growing demand for multiplexed protein detection. In particular, discovery and validation of diseaserelated protein biomarkers require high-throughput detection of many proteins simultaneously. This review will address the complexity of cytokine biology, discuss current and future antibody array technologies and explore their applications in cytokine biomarker discovery and validation for variety of human diseases. Specific examples of these applications will be presented, including the search for cytokine biomarkers related to neurological and neurodegenerative diseases (such as autism and Alzheimer's), immunological disorders (including asthma), and various cancers. ©2012 Bentham Science Publishers.


Kuang Z.,Raybiotech, Inc. | Kuang Z.,South China Biochip Research Center | Wilson J.J.,Raybiotech, Inc. | Luo S.,Raybiotech, Inc. | And 4 more authors.
International Journal of Inflammation | Year: 2015

Asthma is a chronic inflammatory disease of the airways, resulting in bronchial hyperresponsiveness with every allergen exposure. It is now clear that asthma is not a single disease, but rather a multifaceted syndrome that results from a variety of biologic mechanisms. Asthma is further problematic given that the disease consists of many variants, each with its own etiologic and pathophysiologic factors, including different cellular responses and inflammatory phenotypes. These facets make the rapid and accurate diagnosis (not to mention treatments) of asthma extremely difficult. Protein biomarkers can serve as powerful detection tools in both clinical and basic research applications. Recent endeavors from biomedical researchers have developed technical platforms, such as cytokine antibody arrays, that have been employed and used to further the global analysis of asthma biomarker studies. In this review, we discuss potential asthma biomarkers involved in the pathophysiologic process and eventual pathogenesis of asthma, how these biomarkers are being utilized, and how further testing methods might help improve the diagnosis and treatment strain that current asthma patients suffer. © 2015 Zhizhou Kuang et al.


Jiang W.,Raybiotech, Inc. | Mao Y.Q.,Raybiotech, Inc. | Huang R.,Raybiotech, Inc. | Duan C.,Guangzhou University | And 4 more authors.
Journal of Immunological Methods | Year: 2014

Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. © 2013 Elsevier B.V.


Burkholder B.,Raybiotech, Inc. | Huang R.-Y.,Raybiotech, Inc. | Burgess R.,Raybiotech, Inc. | Luo S.,Raybiotech, Inc. | And 8 more authors.
Biochimica et Biophysica Acta - Reviews on Cancer | Year: 2014

Until recently, the intrinsically high level of cross-talk between immune cells, the complexity of immune cell development, and the pleiotropic nature of cytokine signaling have hampered progress in understanding the mechanisms of immunosuppression by which tumor cells circumvent native and adaptive immune responses. One technology that has helped to shed light on this complex signaling network is the cytokine antibody array, which facilitates simultaneous screening of dozens to hundreds of secreted signal proteins in complex biological samples. The combined applications of traditional methods of molecular and cell biology with the high-content, high-throughput screening capabilities of cytokine antibody arrays and other multiplexed immunoassays have revealed a complex mechanism that involves multiple cytokine signals contributed not just by tumor cells but by stromal cells and a wide spectrum of immune cell types. This review will summarize the interactions among cancerous and immune cell types, as well as the key cytokine signals that are required for tumors to survive immunoediting in a dormant state or to grow and spread by escaping it. Additionally, it will present examples of how probing secreted cell-cell signal networks in the tumor microenvironment (TME) with cytokine screens have contributed to our current understanding of these processes and discuss the implications of this understanding to antitumor therapies. © 2014 The Authors.

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