Research progress in the treatment of spinal cord injury with mesenchymal stem cell secretome / 中国组织工程研究

BACKGROUND: Spinal cord injury often leads to patients’ permanent sensory, motor and autonomic dysfunction because of limited inherent regeneration capacity of the central nervous system. Spinal cord injury seriously affects patients’ quality of life but there is still no effective treatment for it. The existing studies suggest that mesenchymal stem cell secretome mediates the main therapeutic effect of cell transplantation and avoids problems such as cellular rejection, so it will become a powerful tool for the treatment of spinal cord injury. OBJECTIVE: To summarize the research progress of mesenchymal stem cell secretome in the treatment of spinal cord injury, and analyze the current problems and future development direction. METHODS: The PubMed database was retrieved with “spinal cord injury, secretome” as the keywords, and Chinese databases include CNKI and Wanfang were retrieved with “spinal cord injury, secretome” as the keywords in Chinese. We retrieved the articles published from January 2013 to January 2020 and excluded the articles that were irrelevant or repetitive. At last 71 articles that met the criteria were included for review. RESULTS AND CONCLUSION: The mesenchymal stem cell secretome is rich in extracellular vesicles and soluble molecules such as growth factors and neurotrophic factors, and it plays prominent place in reducing cell apoptosis, regulating immune response, inhibiting scar formation, promoting nerve regeneration and angiogenesis. Numerous studies have shown that the mesenchymal stem cell secretome can promote nerve regeneration and function recovery after spinal cord injury, and it avoids the disadvantages of cell transplantation. The mesenchymal stem cell secretome will become a reliable method to treat spinal cord injury in the future.

Role of Mesenchymal Stem Cell-Conditioned Medium (MSC-CM) in the Bone Regeneration: A Systematic Review from 2007-2018

Background: The therapeutic value of mesenchymal stem cells (MSCs) in tissue engineering and regenerative medicine is attributable in part to paracrine pathways triggered by several secreted factors secreted into culture media. The secreted factor here is known as the conditioned medium (CM) or secretome. Objectives: This review is aimed to investigate and summarise the in-vitro, pre-clinical in-vivo studies regarding the role of CM-MSC in bone regeneration from 2007 until 2018 Data Sources: A systematic literature search on PubMed, MEDLINE, OVID, Scopus and Cochrane library was carried out by using search terms: Secretome, conditioned medium, mesenchymal stem cell, bone healing, osteogenic, osteogenesis. Methods: A total of 611 articles were reviewed. Ten articles were identified as relevant for this systematic literature review. Results: Three tables of studies were constructed for in vitro studies and in-vivo studies. Conclusion: All of the included in-vitro studies and in-vivo studies have shown a promoting effect of bone regeneration at various stages. Although there are no clinical studies regarding the use of CM-MSC in the human bone regeneration that have been conducted, transplantation of secretome has shown a promising result in the acceleration of bone healing process.

Cell-derived Secretome for the Treatment of Renal Disease

Kidney disease is a major global health issue. Hemodialysis and kidney transplantation have been used in the clinic to treat renal failure. However, the dialysis is not an effective long-term option, as it is unable to replace complete renal functions. Kidney transplantation is the only permanent treatment for end-stage renal disease (ESRD), but a shortage of implantable kidney tissues limits the therapeutic availability. As such, there is a dire need to come up with a solution that provides renal functions as an alternative to the current standards. Recent advances in cellbased therapy have offered new therapeutic options for the treatment of damaged kidney tissues. Particularly, cell secretome therapy utilizing bioactive compounds released from therapeutic cells holds significant beneficial effects on the kidneys. This review will describe the reno-therapeutic effects of secretome components derived from various types of cells and discuss the development of efficient delivery methods to improve the therapeutic outcomes.

Isolation of Secretome with Enhanced Antifibrotic Properties from miR-214-Transfected Adipose-Derived Stem Cells

BACKGROUND: Secretome refers to the total set of molecules secreted or surface-shed by stem cells. The limitations of stem cell research have led numerous investigators to turn their attention to the use of secretome instead of stem cells. In this study, we intended to reinforce antifibrotic properties of the secretome released from adipose-derived stem cells (ASCs) transfected with miR-214. METHODS: We generated miR-214-transfected ASCs, and extracted the secretome (miR214-secretome) from conditioned media of the transfected ASCs through a series of ultrafiltrations. Subsequently, we intravenously injected the miR-214-secretome into mice with liver fibrosis, and determined the effects of miR-214-secretome on liver fibrosis. RESULTS: Compared with that by naïve secretome, liver fibrosis was ameliorated by intravenous infusion of miR-214-secretome into mice with liver fibrosis, which was demonstrated by significantly lower expression of fibrosis-related markers (alpha-smooth muscle actin, transforming growth factor-β, and metalloproteinases-2) in the livers as well as lower fibrotic scores in the special stained livers compared with naïve secretome. The infusion of miR-214-secretome also led to lesser local and systemic inflammation, higher expression of an antioxidant enzyme (superoxide dismutase), and higher liver proliferative and synthetic function. CONCLUSION: MicroRNA-214 transfection stimulates ASCs to release the secretome with higher antifibrotic and anti-inflammatory properties. miR-214-secretome is thus expected to be one of the prominent ways of overcoming liver fibrosis, if further studies consistently validate its safety and efficiency.

Antioxidant action of hypoxic conditioned media from adipose-derived stem cells in the hepatic injury of expressing higher reactive oxygen species

PURPOSE: Almost all liver diseases are known to be accompanied by increased levels of reactive oxygen species (ROS), regardless of the cause of the liver disorder. However, little is known about the role of hypoxic conditioned media (HCM) in the view of pro-oxidative/antioxidative balance. METHODS: Normoxic conditioned media (NCM) and HCM were obtained after culturing adipose-derived stem cells in 20% O₂ or 1% O₂ for 24 hours, respectively. Their effects on the expression of various markers reflecting pro-oxidative/antioxidative balance were investigated in both in vitro (thioacetamide-treated AML12 cells) and in vivo (partially hepatectomized mice) models of liver injury, respectively. RESULTS: HCM treatment induced the higher expression of antioxidant enzymes, such as superoxide dismutase, glutathione peroxidase, and catalase than did NCM in the in vitro model of liver injury. We also found that HCM increased the expression of nuclear factor erythroid 2-related factor (NRF2). The in vivo models of liver injury consistently validated the phenomenon of upregulated expression of antioxidant enzymes by HCM. CONCLUSION: We thus could conclude that HCM provides protection against ROS-related toxicity by increasing the expression of antioxidant enzymes, in part by releasing NRF2 in the injured liver.

El potencial terapéutico del secretoma de las células troncales / The therapeutic potential of stem cells secretome

Resumen Las células madre -o troncales- mesenquimales (CTMs) juegan un papel importante en medicina regenerativa e ingeniería tisular, puesto que estas células indiferenciadas tienen la capacidad de autorrenovarse y de diferenciarse en varios linajes celulares adultos con funciones especializadas diferentes. Estas células cuentan con la capacidad de inmunomodulación y establecimiento celular sin riesgo de generar teratomas, en contraste con las células troncales embrionarias o las células troncales pluripotenciales inducidas. Varios autores también resaltan la capacidad regenerativa de los productos de estas células, conocido como secretoma. Estos productos juegan un papel muy importante en procesos de cicatrización y angiogénesis, por lo que resultan ideales para aplicarlos en terapias basadas en regeneración y reparación de tejidos y órganos. La presente revisión hace un recuento del estado del arte sobre los procesos de inmunomodulación producidos a partir de estos productos celulares.

Abstract Mesenchymal stem cells (MSC's) play an important role in regenerative medicine and tissue engineering, since these undifferentiated cells have the ability to self-renew and differentiate into several adult cell lineages with different, specialized functions. These cells have immunomodulation and homing capacities without the risk of generating teratomas, in contrast to embryonic stem cells or induced pluripotent stem cells. Several authors also highlight the regenerative capacity of products of these cells, known as secretome. These products play a very important role in healing processes and angiogenesis, making them ideal for application in therapies based on regeneration and repair of tissues and organs. The present review makes a recount of the state of the art on the processes of immodulation produced from these cell products.

Prediction and analysis for secretory proteins from Thelazia callipaeda at Genome Scale / 中国人兽共患病学报

We conducted prediction and analysis for secretory proteins from Thelazia callipaeda at Genome Scale based on the previous full genome annotation.The software SignalP,TMHMM,big-PI Predictor,MEME,Protcomp and SecretomeP were combined to process the prediction of the secretome of Thelazia callipaeda.The analyses of secretory proteins by GO function enrichment,KEGG pathway,and statistics of domains were performed.Results showed that totally 259 secretory proteins were found in Thelazia callipaeda genome and the amino acid lengths of secretory proteins were mainly concentrated between 100 to 700 aa exclusively.GO function analysis of secretory proteins indicated that they were enriched in the secreting pathways and in the interactions with host.The results of KEGG metabolism secretory proteins suggested that some of them contributed to drug metabolism and glutathione metabolism.And domain analysis suggested that most of them were glycoside hydrolase,contributing to sugar metabolism.Around 126 secretory proteins had antigenicity of B-cell epitope.In summary,we found that secretory proteins in Thelazia callipaeda were most small proteins,which were involved in sugar metabolism and antioxidative activity,facilitating Thelazia callipaeda to invade the hosts and play a key role in the parasitic course.

Current Understanding of Stem Cell and Secretome Therapies in Liver Diseases

Liver failure is one of the main risks of death worldwide, and it originates from repetitive injuries and inflammations of liver tissues, which finally leads to the liver cirrhosis or cancer. Currently, liver transplantation is the only effective treatment for the liver diseases although it has a limitation due to donor scarcity. Alternatively, cell therapy to regenerate and reconstruct the damaged liver has been suggested to overcome the current limitation of liver disease cures. Several transplantable cell types could be utilized for recovering liver functions in injured liver, including bone marrow cells, mesenchymal stem cells, hematopoietic stem cells, macrophages, and stem cell-derived hepatocytes. Furthermore, paracrine effects of transplanted cells have been suggested as a new paradigm for liver disease cures, and this application would be a new strategy to cure liver failures. Therefore, here we reviewed the current status and challenges of therapy using stem cells for liver disease treatments.

Caracterização do secretoma de cepas de Xylella fastidiosa / Characterization of the secretome Xylella fastidiosa strains

As doenças causadas pelo fitopatógeno Xylella fastidiosa, uma bactéria Gram-negativa, devem-se aos seus múltiplos fatores de virulência, tais como formação de biofilme, secreção de enzimas de degradação da parede celular do xilema (CWDE), expressão de proteínas de adesão e produção de vesículas de membrana externa (OMVs). Esses fatores de virulência são controlados por uma via de sinalização mediada por DSF (fatores de sinalização difusíveis de natureza lipídica) e relacionada com percepção de quórum. Nesse trabalho, tivemos como objetivo ampliar a caracterização do secretoma de cepas selvagens e mutantes de X. fastidiosa para evidenciar proteínas e metabólitos potencialmente associados à adaptação ao hospedeiro, virulência e patogenicidade. Desenvolvemos, paralelamente, três estudos empregando como abordagens metodológicas a proteômica, a metabolômica e a transcritômica. No primeiro estudo, comparamos o secretoma (exoproteoma) da cepa Temecula1 selvagem (WT) e do mutante no gene da sintase de DSF (ΔrpfF), o qual exibe fenótipo de hipervirulência em videiras. A este estudo associamos a comparação dos transcritomas dessas cepas. Os resultados mostraram que, mesmo no cultivo in vitro, X. fastidiosa expressa e secreta fatores de virulência previamente conhecidos (lipases-esterases e proteases), além de toxinas (microcinas) que, supostamente, teriam papel de controlar bactérias competidoras pelo mesmo nicho. No segundo estudo caracterizamos a composição de OMVs secretadas no cultivo in vitro por X. fastidiosa Fb7 e 9a5c (cepas isoladas de laranjeiras) e Temecula1 (cepa isolada de videira). Demonstramos que Fb7 produz até 57% mais OMVs que 9a5c e Temecula1 e identificamos um total de 202 proteínas distintas nas OMVs produzidas pelas 3 cepas, ampliando consideravelmente o número de proteínas secretadas por meio de OMVs descrito, até então, para X. fastidiosa. Entre as proteínas enriquecidas, citamos adesinas afimbriais, porinas, lipoproteínas, hidrolases (lipases/esterases, proteases e peptidases) e uma pectina-liase putativa. Destacamos a detecção da enzima L-ascorbato oxidase nas OMVs e sugerimos que esta enzima poderia atuar na depleção do ascorbato produzido pelo hospedeiro vegetal. Além disso, demonstramos, pela primeira vez, que OMVs de X. fastidiosa transportam ácidos graxos da família DSF, sugerindo um papel adicional para OMVs nesse fitopatógeno. Finalmente, no terceiro estudo verificamos alterações relevantes no perfil de metabólitos secretados por X. fastidiosa em resposta a sua interação com metabólitos secretados por Burkholderia phytofirmans, proposta como uma cepa para o biocontrole da doença de Pierce de videiras. Confirmamos que o sobrenadante de B. phytofirmans possui um composto de natureza apolar que induz a formação de biofilme em X. fastidiosa, contudo ainda não foi possível decifrar a natureza química deste composto

The diseases caused by the phytopathogen Xylella fastidiosa, a Gram-negative bacterium, are due to multiple virulence factors, such as biofilm formation, secretion of xylem cell wall degradation enzymes (CWDE), expression of adhesion proteins and production of outer membrane vesicles (OMVs). These virulence factors are controlled by a DSF (diffusible signaling factors of a lipidic nature) mediating signaling pathway and related to quorum sensing perception. In this work, we aimed to extend the characterization of the secretoma of wild type and mutants strains of X. fastidiosa to uncover proteins and metabolites potentially associated to host adaptation, virulence and pathogenicity. We developed three studies in parallel using proteomics, metabolomics and transcriptomics as methodological approaches. In the first study, we compared the secretome (exoproteome) of the wild type strain Temecula1 (WT) and of DSF synthase mutant (ΔrpfF) which exhibits hypervirulence phenotype in grapevines. We also compared the transcriptomes of these strains. Our results showed that, even in in vitro culture, X. fastidiosa expresses and secretes previously known virulence factors (lipasesesterases and proteases), as well as toxins (microcins) that might play a role in controlling competing bacteria in the same niche. In the second study, we characterized the composition of OMVs secreted by in vitro cultures of X. fastidiosa Fb7 and 9a5c (strains isolated from orange trees) and Temecula1 (strain isolated from grapevine). We have shown that Fb7 produces up to 57% more OMVs than the 9a5c and Temecula1. Moreover we identified a total of 202 distinct proteins in the OMVs produced by these three strains, increasing considerably the number of OMVs secreted proteins so far described for X. fastidiosa. Among the proteins enriched in OMVs, we point out afimbrial adhesins, porins, lipoproteins, hydrolases (lipases/esterases, proteases and peptidases) and a putative pectin-lyase. We highlight the detection of the enzyme L-ascorbate oxidase in the OMVs and we suggest that this enzyme could act in the depletion of ascorbate produced by the plant host. In addition, we have demonstrated, for the first time, that X. fastidiosa OMVs transport fatty acids from the DSF family, suggesting an additional role for OMVs in this phytopathogen. Finally, in the third study we verified relevant changes in the profile of metabolites secreted by X. fastidiosa in response to the interaction with metabolites secreted by Burkholderia phytofirmans that has been sugested as a biocontrol strain for Pierce's disease in grapevines. We confirm that the B. phytofirmans supernatant has a non-polar compound that induces biofilm formation in X. fastidiosa, but it has not yet been possible to elucidate the chemical nature of this compound

Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells / 대한악안면성형재건외과학회지

BACKGROUND: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor (TGF)-β1, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. METHODS: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. RESULTS: The concentrations of IGF-1, VEGF, and TGF-β1 in MSC-CM were 1515.6 ± 211.8 pg/mL, 465.8 ± 108.8 pg/mL, and 339.8 ± 14.4 pg/mL, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. CONCLUSIONS: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

Liver Regenerating Potential of the Secretome Obtained from Adipose-derived Stem Cells Cultured under the Hypoxic Environment / 대한이식학회지

BACKGROUND: The stem cell-derived secretome has received considerable attention as an alternative to stem cells for therapeutic applications. However, establishing optimal culture conditions is key to obtaining appropriate secretome contents. Here, the optimal culturing environment for achieving a high-efficiency secretome was determined via hypoxic preconditioning of human adipose-derived stem cells (ASC). METHODS: Normoxic conditioned media (NCM) and hypoxic conditioned media (HCM) were obtained after culturing human ASCs under normoxia (20% O2) or hypoxia (1% O2), respectively. Subsequently, both normal and thioacetamide-induced hepatotoxic hepatocytes were treated with NCM or HCM. In addition, partially hepatectomized mice were infused with control saline, NCM, and HCM. The effects on liver regeneration and serum transaminases levels were then compared. RESULTS: Hypoxic preconditioning significantly increased mRNA expression of proinflammatory cytokines (interleukin-6 and tumor necrosis factor-α) and growth factors (hepatocyte growth factor and vascular endothelial growth factor). In both normal and thioacetamide-induced hepatotoxic hepatocyte (alpha mouse liver 12 [AML12]) cell lines, HCM treatment resulted in the highest cell viability (122% and 95%, respectively), followed by NCM (111% and 78%, respectively). In addition, intravenous administration of HCM to partially hepatectomized mice resulted in substantially enhanced liver regeneration compared with the NCM group (P<0.05). CONCLUSIONS: Taken together, the secretome obtained from ASC with hypoxic preconditioning showed potential to alleviate liver damage both in vitro and in vivo. Hypoxic culture of ASC is expected to play an important role in regenerative medicine by inducing secretome production that is beneficial for improving liver regeneration.

Differential toxicological endpoints of di(2-ethylhexyl) phthalate (DEHP) exposure in MCF-7 and MDA-MB-231 cell lines: Possible estrogen receptor α (ERα) independent modulations.

Wide spread use of Di-(2-ethylhexyl) phthalate (DEHP) has made it a ubiquitous contaminant in today’s environment, responsible for possible carcinogenic and endocrine disrupting effects. In the present investigation an integrative toxico-proteomic approach was made to study the estrogenic potential of DEHP. In vitro experiments carried out with DEHP (0.1-100 μM) induced proliferations (E-screen assay) in human estrogen receptors-α (ERα) positive MCF-7 and ERα negative MDA-MB-231 breast cancer cells irrespective of their ERα status. Further, DEHP suppressed tamoxifen (a potent anti-breast cancer drug) induced apoptosis in both cell types as shown by flowcytometric cell cycle analysis. Label-free quantitative proteomics analysis of the cell secretome of both the cell lines indicated a wide array of stress related, structural and receptor binding proteins that were affected due to DEHP exposure. The secretome of DEHP treated MCF-7 cells revealed the down regulation of lactotransferrin, an ERα responsive iron transport protein. The results indicated that toxicological effects of DEHP did not follow an ERα signaling pathway. However, the differential effects in MCF-7 and MDA-MB-231 cell lines indicate that ERα might have an indirect modulating effect on DEHP induced toxicity.

Progress in pathogenic secretome research / 国际儿科学杂志

Secretome includes all the gene products secreted by a cell. The development of proteomicbased approaches and genome-based computational prediction technology have gradually revealed secretome plays an important role in pathogen infection recently. Secretome facilitates pathogen to proliferate and invade the host by inhibiting the host's immune system, resisting acidic PH conditions, snatching nutrition and so on.Identification of pathogenic secretome helps discovering new drug targets and biomarkers, and supporting the diagnosis and treatment.

Análise proteômica comparativa dos componentes da superfície celular e do secretoma do Aspergillus fumigatus / Comparative proteomic analysis of the components of the cell surface and secretome of Aspergillus fumigatus

Aspergillus fumigatus é o principal agente etiológico da aspergilose invasiva, infecção fúngica oportunista com altas taxas de mortalidade afetando, principalmente, pacientes com neutropenia profunda e prolongada. Durante o processo de invasão e disseminação características desta infecção sistêmica, os conídios do fungo inalados e não eliminados pelas células do sistema imune inato diferenciam-se em hifas que, por sua vez, são angioinvasivas. Pouco se conhece sobre as moléculas da parede celular envolvidas na patogênese do A. fumigatus e/ou secretadas por este patógeno. Neste contexto, este trabalho procura ampliar o entendimento desta doença através do estudo de proteínas diferencialmente expressas na superfície de A. fumigatus durante a morfogênese. Foi utilizada uma abordagem proteômica e foram estudados extratos de superfície de células de A. fumigatus em diferentes estágios durante o processo de filamentação. Estas células foram denominadas, de acordo com o tempo de cultivo e a morfologia, como: TG6h (tubo germinativo), H12h ou H72h (hifas). As proteínas de superfície celular foram extraídas, a partir de células intactas, por tatamento brando com o agente redutor DTT (ditiotreitol). Observou-se que o perfil funcional das proteínas expressas por H12h e H72h foi similar, com exceç~çao de proteínas relacionadas à resposta ao estresse, enquanto o perfil para TG6h apresentou diferenças significativas para vários grupos funcionais de proteínas quando comparado às hifas. Desta forma, foram realizados experimentos de proteômica diferencial entre tubo germinativo (TG6h) e a hifa madura (H72h), pela técnica de DIGE (differential gel electrophoresis). Os resultados revelaram que entre as proteínas diferencialmente expressas, aquelas relacionadas às vias de biossíntese e outras denominadas multifuncionais encontram-se superexpressas em TG6h. Em relação às proteínas de resposta a estresse, observou-se que algumas HSPs eram mais expressas neste morfotipo...

Aspergillus fumigatus is the main etiologic agent of invasive aspergillosis (IA), a opportunistic a life-threatening disease for immunocompromised hosts, especially those with acute and prolonged neutropenia. During the invasion and dissemination, which occurs in this systemic infection, the A. fumigatus conidia, after its inhalation, germinates into angioinvasive hyphae in case the innate immune response fails in eliminate these cells. Little is known about the cell wall molecules and/or the secreted proteins involved on the A. fumigatus pathogenesis, at this context the present work aims to amplify the knowledge about the aspergillosis by studying the differentially surface proteins of A. fumigatus during the filamentation process. These cells were denominated according to their morphology and their growtn time as: TG6h (germ tubes), H12h and H72h (hyphae). The surface proteins were mildly extracted from intact cells using the reducing agent DTT (dithiothreitol). The functional profile of the H12h and H72h were similar except for the stress response proteins, while the TG6h presented significant differences for several functional groups. On this base, the DIGE (differential gel electrophoresis) was performed using the surface extracted proteins of the germ tubes (TG6h) and mature hyphae (H72h) cells. The results indicate that multiple functional proteins and proteins related to the biosynthesis pathways were overexpressed at TG6h. Some stress response proteins as the HSPs were overexpressed on this morphotype while the MnSOD, oxidative stress responsive protein, was most abundant at the hyphae. PhiA, an integrant protein of the cell wall, was the only protein with a secretion signal sequence. All other proteins identified on the cell surface lack an identifiable secretion sign, and are denominated atypical proteins. The plasma membrane integrity was verified after the mild extraction using DTT, and also the biotinylation of the cell extracted proteins...

Serum HSP90α in the clinical stage of non-small cell lung cancer / 国际外科学杂志

Objective To investigate whether HSP90α could be a sensitive and specific serum biomarker for the diagnosis and progression of lung cancer. Methods In the present study, different secretomic analy-ses on the two human lung adenocarcinoma cell lines CL1-0 and CL1-5 with low and high metastatic poten-tial, respectively, were performed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ma-trix-assisted laser desorption/ionization time-of-flight mass spectrometry. The candidate biomarker was con-firmed by Western blotting, and was further analyzed in 224 serum samples including 141 lung cancer, 37 benign pulmonary diseases, as well as 46 healthy individuals using ELISA assay. Results HSP90α was sig-nificantly upregulated in the CM of CL1-5 cells. It was found that the levels of HSP90α were specifically ele-vated in the sera of non-small cell lung cancer compared with other groups. At the cut-off point 0.535 on the receiver operating oharacteristie curve, HSP90α could comparatively discriminate lung cancer from benign lung disease and healthy control groups with sensitivity of 0. 817, specificity 0. 919 and total accuracy 80. 14%. Conclusion HSP90α may be a potential useful serum biomarker for discriminating lung cancer from benign lung diseases and healthy individuals and staging of non-small cell lung cancer.

Screening of EGFR-regulated Secreted Proteins in Human NPC Cell Line CNE2 / 生物化学与生物物理进展

In order to screen EGFR-regulated secreted proteins in human nasopharyngeal carcinoma(NPC), and to reveal the role and mechanism of epidermal growth factor receptor(EGFR) in the pathogenesis of NPC. NPC cell line CNE2 cells were cultured in serum-free medium and stimulated by transforming growth factor-? (TGF-?) for 24 h in experimental group. Control CNE2 cells were cultured at the same condition but without TGF-? stimulation. The culture medium of control and experimental cells was desalted and concentrated through ultrafiltration to prepared the total secreted proteins. Two-dimensional gel electrophoresis (2-DE) was used to separate the secreted proteins of control and experimental cells, PDQuest software was applied to analyze 2-DE images, and the differential protein spots between the control and experimental cells were identified by desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The 2-DE patterns of the secreted proteins of TGF-? stimulated and un-stimulated CNE2 cells were established, 22 differential proteins spots between the two groups of cells were found, and 8 non-redundant proteins were identified with MALDI-TOF-MS, the functions of which were involved in invasion, metastasis, apoptosis and proliferation of cancer cells. The data will be valuable for further to study the role and mechanism of EGFR in the pathogenesis of NPC.