Research progress on the role of MAGE-A3 in the occurrence and development of esophageal cancer / 中国肿瘤生物治疗杂志

@#[摘 要] 食管癌（EC）是临床常见的恶性肿瘤之一，近年来免疫治疗的应用使EC患者的预后得到了改善，但患者对免疫治疗的应答有着明显的异质性，且欠缺有效的疗效预测生物标志物，无法精准筛选获益人群，存在原发性耐药，因此需要寻找新型免疫治疗靶点使更多患者获益。黑色素瘤相关抗原A3（MAGE-A3）是一种癌-睾丸抗原，在正常组织中几乎不表达，而在多种恶性肿瘤组织中呈现高表达，参与肿瘤细胞的增殖、侵袭和转移等多种生物学过程。MAGE-A3基因具有限制性表达和良好的免疫原性，是良好的EC免疫治疗靶点，对EC的诊断、治疗和预后判断等具有良好的应用价值。由于MAGE-A3基因在EC组织中高表达，并与不良预后相关。目前针对MAGE-A3基因靶点的肿瘤疫苗和过继性细胞免疫治疗等疗法均展现出良好的应用前景。

MAGED4B Promotes Glioma Progression via Inactivation of the TNF-α-induced Apoptotic Pathway by Down-regulating TRIM27 Expression / 神经科学通报·英文版

MAGED4B belongs to the melanoma-associated antigen family; originally found in melanoma, it is expressed in various types of cancer, and is especially enriched in glioblastoma. However, the functional role and molecular mechanisms of MAGED4B in glioma are still unclear. In this study, we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue, and the level was positively correlated with glioma grade, tumor diameter, Ki-67 level, and patient age. The patients with higher levels had a worse prognosis than those with lower MAGED4B levels. In glioma cells, MAGED4B overexpression promoted proliferation, invasion, and migration, as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide. On the contrary, MAGED4B knockdown in glioma cells inhibited proliferation, invasion, and migration, as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide. MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain. The interaction between MAGED4B and tripartite motif-containing 27 (TRIM27) in glioma cells was detected by co-immunoprecipitation assay, which showed that MAGED4B was co-localized with TRIM27. In addition, MAGED4B overexpression down-regulated the TRIM27 protein level, and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine (MG132), an inhibitor of the proteasome. On the contrary, MAGED4B knockdown up-regulated the TRIM27 level. Furthermore, MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132. Accordingly, MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7 (USP7) involved in the tumor necrosis factor-alpha (TNF-α)-induced apoptotic pathway. These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1 (RIP1)-dependent TNF-α-induced apoptotic pathway, which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.

Reporte de caso: síndrome de Schaaf Yang / Case report: Schaag Yang síndrome

RESUMEN Introducción: El síndrome Schaaf Yang (SHFYNG) constituye un desorden multisistémico caracterizado por un grupo de signos y síntomas relacionados con alteraciones genéticas, congénitas y de expresión clínica multivariable. Fue descrito por primera vez por el Dr. Schaaf y la Dra. Yaping, profesores de Genética Molecular y Humana de la Universidad de Houston y Baylor, respectivamente, en 2013 (1). El síndrome SHFYNG tiene una herencia autosómica dominante con una mutación presente en el alelo paterno, ya que el gen MA-GEL2 tiene una impronta materna y solo se expresa el alelo paterno. A diferencia de otras patologías autosómicas dominantes clásicas, el síndrome SHFYNG puede saltar varias generaciones siempre que la mutación resida en el cromosoma materno. Presentación del caso. Preescolar femenina, con antecedente de estancia en UCIN por hipotonía neonatal y pobre succión, bronquitis y neumonía. Su fenotipo está caracterizado por facies hipotónicas, frente prominente, epicanto interno, pómulos prominentes, puente nasal bajo, nariz ancha, labio superior delgado, orejas aladas, cuello corto y obesidad central. Presenta retraso en el neurodesarrollo, lenguaje y psicomotor. Estudios genéticos: cariotipo 46,XX e hibridación genómica comparativa con patrón genómico normal, sexo femenino, en exoma trío se identifica una variante patogénica: c.1996dupC (p.Gln666Profs*47) en el gen MAGEL2 asociada con SHFYNG. Conclusión. Se informa el primer reporte de este síndrome a nivel nacional con una incidencia mundial muy baja, estimándose aproximadamente <1/1.000.000 de nacidos vivos, lo que permite ampliar el conocimiento y sospechar patologías de difícil diagnóstico como esta.

ABSTRACT Introduction: Schaaf Yang Syndrome (SHFYNG) is a multisystemic disorder characterized by a group of signs and symptoms related to genetic, congenital, and multivariate clinical alterations. It was first described by Dr. Schaaf and Dr. Yaping, professors of Molecular and Human Genetics at the University of Houston and Baylor, respectively, in 2013 (1). SHFYNG has an autosomal dominant inheritance with a mutation located in the paternal allele, since the MAGEL2 gene has a maternal imprint and only the paternal allele is expressed. Unlike other classic autosomal dominant pathologies, SHFYNG syndrome can skip several generations, as long as the mutation resides on the maternal chromosome. Presentation of the case: Female preschooler, with a history of stay in the Neonatal Intensive Care Unit, due to neonatal hypotonia and poor suction, bronchitis, and pneumonia. Her phenotype is distinguished by hypotonic facies, prominent forehead, internal epican-thus, prominent cheekbones, low nasal bridge, broad nose, thin upper lip, winged ears, short neck, and central obesity. She presents neurodevelopmental, language, and psychomotor delay. Genetic studies: 46,XX karyotype, comparative genomic hybridization: normal genomic pattern, female sex, trio exam a pathogenic variant c.1996dupC (p.Gln666Profs*47) in the MAGEL2 gene associated with SHFYNG syndrome. Conclusion: It is reported to be the first national report of this syndrome, with a very low worldwide incidence, estimating approximately <1 / 1,000,000 live births, which allows us to expand knowledge and suspect difficult-to-diagnose pathologies like this one.

Expression and clinical significance of MAGE-C1 in breast cancer tissues / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of MAGE-C1 (melanoma-associated antigen-C1) in breast cancer tissues and its correlation with clinicopathological features and prognosis of breast cancer patients. Methods: Breast cancer tissues, normal breast tissues and benign breast lesion tissues (60 samples for each) were collected from the Fourth Hospital of Hebei Medical University during January 2008 and December 2008.The mRNA and protein expressions of MAGE-C1 in three types of breast tissues were detected by RT-PCR and immunohistochemistry, and their correlation with clinicopathological parameters and prognosis of breast cancer patients were also analyzed. DNA methylase inhibitor 5-aza-2'-deoxycytidine (5-Aza-CdR) and histone deacetylase inhibitor trichostatin A (TSA) were used to treat breast cancer MDA-MB-231 and MCF-7 cells, and RT-PCR was used to determine the changes in mRNA expression of MAGE-C1 after drug treatment. Results: The positive expression rate of MAGE-C1 mRNA and protein in breast cancer tissues were 43.3% (26/60) and 38.3% (23/60), respectively; and the mRNA and protein expressions of MAGE-C1 were all negative in normal breast tissues and benign breast lesion tissues. MAGE-C1 expression was positively associated with high tumor grade (χ2 =6.233, P<0.05). Recurrence-free survival (RFS) of patients with negative MAGE-C1 expression was significantly longer than those patients with positive MAGE-C1 expression (χ 2 =4.213, P<0.05). MAGE-C1 expression (HR=3.980, P<0.05) and clinical stage (HR=3.637, P<0.05) could be used as independent prognostic factors for breast cancer patients. 5-Aza-CdR and/or TSA treatment had no significant influence on MAGE-C1 gene expression (P>0.05). Conclusion: MAGE-C1 is a tumor-specific antigen and its expression is associated with poor prognosis of breast cancer patients.

Osteosarcoma of the jaw - experience at the Medical University Vienna and comparative study with international tumor registries

OBJECTIVES: Osteosarcoma of the jaw (OSAJ) is fundamentally different in clinical practice from its peripheral counterparts. Studies are difficult to conduct due to low incidence rates. The primary aim of this study was to provide for the first time a comprehensive retrospective analysis of the treatment concepts and outcome data of OSAJ patients treated at the University Hospital Vienna and to compare these with two recently published studies on OSAJ. The clinical study was accompanied by a biomarker study investigating the prognostic relevance of melanoma-associated antigen-A (MAGE-A) in OSAJ specimens. METHOD: Eighteen patients were included, and their outcomes were compared to published data. Immunohistochemistry was performed with mouse monoclonal antibodies against MAGE-A. Survival rates were estimated by the Kaplan-Meyer method. The log-rank test was used to analyze potential prognostic parameters. Fisher's exact test was performed to define the significant differences between the survival rates of the current study and the DOESAK registry. RESULTS: Disease-specific survival was 93.8% after five and 56.3% after ten years. The development of metastases (p=0.033) or relapse (p=0.037) was associated with worsened outcomes in our group as well as in the comparative group. Despite the different treatment concepts of the study groups, survival rates were comparable. MAGE-A failed to show prognostic relevance for OSAJ patients. CONCLUSIONS: Uncertainties about the optimal treatment strategies of OSAJ patients will currently remain. Thus, prospective studies of OSAJ are needed but are only feasible in a multicenter study setting, conducted over a prolonged time period.

Expressions and clinical significances of MAGE-A9, MAGE-A11 and Ki67 in laryngeal squamous cell carcinoma / 中国肿瘤生物治疗杂志

@# Objective: To explore the expressions of melanoma antigen (MAGE) -A9, -A11 and Ki67 in laryngeal squamous cell carcinoma (LSCC) tissues, and to analyze their correlation with clinicopathological features and the prognosisof LSCC patients. Methods: A total of 73 pairs of LSCC tissuesand corresponding para-cancerous tissues resected from LSCC patients, who were treated at the Fourth Hospital of Hebei Medical University from 2012 to 2014,were collected for this study. At the same time, testicular tissues from 3 patients with prostate cancer after castration were selected as positive control. The protein expressions of MAGE-A9, MAGE-A11 and Ki67 in LSCC tissues and its para-cancerous tissues were detected by immunohistochemistry. Results: The expression rates of MAGEA9, MAGE-A11 protein and Ki67 in LSCC tissues were 47.94% (35/73), 49.32% (36/73) and 46.58% (34/73) respectively, which were significantly higher than those in para-cancerous tissues. The protein expressions of MAGE-A9 and MAGE-A11 were correlated with clinical stage and lymphatic metastasis of LSCC (P<0.05). The expression of Ki67LI was correlated with tumor size, clinical stage and lymphatic metastasis of LSCC (P<0.05). The correlation analysis showed that the expressions of MAGE-A9 and MAGE-A11 were positively correlated with Ki67 (r=0.258, P＝0.027; r=0.672, P=0.001). Kaplan-Meier survival curve analysis showed that the survival rates of patients with high expression of MAGE-A9 protein (P=0.009), MAGE-A11 protein (P=0.031) and Ki67LI (P=0.040) were significantly lower than those with low expressions. And the survival time of patients with both high expressions of MAGE-A9 and Ki67LI (P=0.001) or both high expressions of MAGE-A11 and Ki67 (P=0.001) was significantly shorter than that of patients with low expression (both or single). Univariate and multivariate Cox regression analysis further indicated that MAGE-A9 protein (P=0.028) and MAGE-A11 protein (P=0.042) were independent prognostic factors for overall survival of LSCC patients. Conclusion: MAGE-A9, MAGE-A11 and Ki67 are tumor-associated antigens of LSCC, which can be used as prognostic indicators for LSCC.

Screening and identification of MAGE-A11 related genes based on DNAmicroarray / 中国肿瘤生物治疗杂志

@# Objective: To screen related genes of melanoma-associated antigen-A11 (MAGE-A11) in breast cancer cells based on highthroughput DNAmicroarray technology, and to validate from the aspects of quantity and function. Methods: DNAmicroarray was used to screen the differently-expresseddown-stream mRNAs of MAGE-A11 in breast cancercelllines (MCF-7, MDA-MB-231 and BT-549). Cluster analysis was applied on representative genes and quantitative RT-PCR was used to validate. CCK-8, scratch wound healing assay and Transwell assaywere used to detect the effect of MAGE-A11 on the proliferation,migration and invasion of breast cancer cells. Results: Over-expression of MAGE-A11 caused the differential expression of 1608 down-stream genes in 3 breast cancer cell lines, which was associated with various cell functions such as protein ubiquitination,cell proliferation and apoptosis, tumor invasion and metastasis.qRT-PCR validated that the expression of ZNF-451, CENPTJ, CDK13, API5 and LMO7, which were highly expressed in microarray, were also significantly higher than those in control group (P<0.01);in addition, SHPRH, PML, MARK2, LIMA1 and ANGPTL4, which were low-expressed in microarray, were also significantly lower than those in control group (P<0.01). MAGE-A11transfection directly increased the proliferation of breast cancer MCF-7, MDA-MB-231 and BT-549 cells at 72 h (all P<0.01); compared with control group after transfectionexhibited obvious wound healing at 48 h (P<0.05 or P<0.01) and significantly increased trans-membrane cell numbers (all P<0.01). Conclusion: Many differentially expressed genes related to ubiquitination, cell proliferation and apoptosis, tumor invasion and migration were screened in MCF-7, MDA-MB-231 and BT-549 breast cancer cells. Among them, 10 typical differentially expressed genes were identified in terms of quantity and function.

Inhibitory effects of DC vaccine sensitized with MAGE-3 combined with CpG ODN on bladder cancer BIU-87 cells / 中国肿瘤生物治疗杂志

@#Objective: To investigate the role of CpG ODN (CpG oligodeoxynucleotide) adjuvant in enhancing the anti-bladder cancer response induced by MAGE-3 (melanoma antigen gene -3) antigen and its molecular mechanism. Methods: Mononuclear cells were isolated from HLA-A2 type peripheral blood of healthy donors by Ficoll method to prepare mature DC by conventional means. DC surface markers were detected by flow cytometry. MTT assay was used to detect the promotion effect of DCs sensitized by different means (MAGE-3, CpGODN, MAGE-3+CpG ODN, irrelevant control antigen) on the proliferation of T lymphocytes and the killing effect of CTL on BIU-87 tumor cells. The tumor mass of nude mice bearing BIU-87 bladder cell xenograft were examined on Day 7 and 11 after CpG ODN+MAGE-3 sensitized DC treatment. The expression of Bcl-2/Bax protein was detected by Western blotting while the proliferation level of xenograft cells was detected by MTT assay. Results: DCs sensitized by CpG ODN combined with MAGE-3 antigenic peptides could promote the proliferation of T lymphocytes and significantly enhance the killing effect of CTLon target BIU-87 cells (P< 0.05). Compared with other sensitized DCs, in vivo experiments showed that 7 and 11 days after treatment, both the tumor volume and weight were significantly reduced (all P<0.05), and the proliferation ability of xenograft tumor was decreased (P<0.05). Compared with other sensitization means, CpG ODN+MAGE-3 especially exhibited obvious inhibitive effect on tumor growth on Day 11, and significantly promoted the proliferation of splenic monocytes of tumor bearing mice (P<0.01); moreover, Bcl-2 expression in xenograft tissues significantly decreased（P<0.01）while Bax expression significantly increased（P<0.05 or P<0.01）on Day 3 after treatment. Conclusion: CpG ODN can promote the inhibitory effect of MAGE-3 sensitized DC on bladder cancer BIU-87 cells, which will provide experimental basis for clinical application of DC vaccine in bladder cancer treatment.

Expression and significance of MAGE-A9 in hepatocellular carcinoma / 肿瘤研究与临床

Objective To detect the expression of MAGE-A9 in human hepatocellular carcinoma (HCC), and to investigate the association between expression of MAGE-A9 and the clinicopathological characteristics of HCC.Methods Reverse transcription-polymerase chain reaction (RT-PCR), one-step quantitative-PCR (qPCR) and immunohistochemistry (IHC) methods were performed to characterize the expression of MAGE-A9 in HCC cell lines and tissues.Kaplan-Meier survival analysis and Cox regression were employed to evaluate the prognosis of 100 HCC patients.Results The expression of MAGE-A9 mRNA in HCC (4.44±0.342) was significantly higher than that in non-cancerous cells and tissues (1.73±0.178) (P ＜ 0.05).The expression level of the MAGE-A9 protein in HCC was related to the pathological grade (P =0.003), portal vein invasion (P =0.001), distant metastasis (P =0.022) and TNM stage (P =0.005).Cox regression analysis revealed that MAGE-A9 expression is an independent prognostic factor for disease-free survival (P =0.006) and overall survival (P =0.022).Conclusion MAGE-A9 is a valuable prognostic biomarker for HCC patients, and its high expression suggests unfavorable survival outcomes in HCC patients.

Effects of subcellular localization of neurotrophin receptor p75 interacting MAGE homologue on the epithelial to mesenchymal transition and radioresistance of esophageal cancer cells / 中华放射医学与防护杂志

Objective To investigate the role of NRAGE subcellular localization in the EMT and radioresistance of esophageal cancer cells.Methods EMT model cells were established by the treatment of TE13 cells with TGF-β1.To verify the establishment of EMT model and the phenotype of EMT-like TE13R120 cells,EMT marker mRNA and protein were detected by Real-time PCR and Western blot,respectively.Real-time PCR was also used to detect the expression of NRAGE mRNA in three groups.Total NRAGE protein,cytoplasm protein and nuclear protein were measured by Western blot.Results It was found that TGF-β1 could induce morphological alterations of TE13 cells from epithelial to mesenchymal and change the expressions of EMT maker E-cadherin and vimentin (t =13.56,-232.84,P ＜ 0.05),indicating the successful establishment of EMT model cells.Similar expression trends of EMT makers were observed in TE13R120cells (t=15.84,-54.54,P＜0.05).NRAGE mRNA (t=-8.73,-5.62,P＜ 0.05) and total protein in both EMT model cells and TE13R120 cells were higher than that in TE13 cells,especially for the nuclear proteins.However,no differences in NRAGE cytoplasm protein expression were found among the three groups.In addition,there were also no difference of NRAGE mRNA (t =-0.88,P ＞0.05),cytoplasm and nuclear protein between TE13R120 cells and EMT model cells.Conclusions The radioresistant cell line TE13R120 has the EMT-like phenotype that may cause cell radioresistance by changing the subcelluar localization of NRAGE.

Quantitative expression of Melanoma-associated antigen MAGE-E1 mRNA in glioma / 中国免疫学杂志

Objective:To investigate the quantitative expression of Melanoma-associated antigen MAGE-E1 mRNA in glioma, and explore its potential for immunotherapy in glioma.Methods:To establish a quantitative real-time polymerase chain reaction ( qRT-PCR) method to quantitatively determine MAGE-E1 mRNA in glioma.A total of 47 human glioma and 14 normal brain tissue specimens were analyzed.MAGE-E1 mRNA was normalized to HPRT1, a housekeeping gene.MAGE-E1/HPRT1 was used to evaluate the expression level of MAGE-E1 mRNA.Results:High-level expression of MAGE-E1 was observed in 7.1%of normal brain and 66.0%glioma tissues,which shown significant difference with P<0.05.There was irrelevant between the expression of MAGE-E1 mRNA and clinicopathogical parameters ,such as gender ,age,histological subtype and grade.Conclusion:With higher expression of MAGE-E1 in glioma tissues,it might be a potential target for immunotherapy of glioma.

Lipoxygenase-mediated oxidative metabolism and oxidative stress / 中国药理学与毒理学杂志

There is a close relationship between oxidative stress induced tissue damage and many diseases.As a multi-function oxidase,lipoxygenase can generate reactive oxygen species (ROS)by oxidatively metabolizing various endogenous and exogenous chemicals such as drugs and environmental pollutants.Lipoxygense can catalyze endogenous che mical reaction,such as arachidonic acid and pro-duce ROS.Lipoxygenase can be also activated in other ways to generate ROS by activating signal trans-duction.Moreover,some exogenous chemicals can be metabolized into highly reactive radical intermedi-ates by lipoxygenase,inducing ROS generation.The accumulation of intracellar ROS can damage intra-cellar redox balance and induce oxidative stress.That may be one of the possible toxic effect mecha-nis ms of che mical agents.

Protection of vitamin C on the cardiac injury induced by nano-titanium dioxide in mice / 中国药理学与毒理学杂志

OBJECTIVE To observe the protection of vitamin C on the cardiac injury induced by 50 nm titanium dioxide inmice.METHODS Kunming mice were ad mistered by ig of vitamin C 100,200 and 400 mg·kg -1 for 2 d.And then the mice were ad mistered by ig of nano-TiO2 2 g·kg -1 and vitamin C (100.0,200.0 and 400.0 mg·kg -1 )for 3 d,the interval of treatment with nano-TiO2 and vitamin C was 4 h.The mice were scarified 24 h later after the last ad ministration.Electrocardiogra m (ECG)was determinated by physiological recorder.The myocardial enzy mes activities in serum and superoxide dismutase (SOD)and glutathione peroxidase(GSH-Px)activities in serum and myocardial tissue were determinated by bioche mical method.Cometassay was used to detect the DNA da mage of the heart. Heart tissue was used for histopathological exa mination by HE staining.RESULTS Co mpared with the control,ECG showed higher S-T and T-wave a mplitude of nano-TiO2 2 g·kg -1 (P<0.05).The myocar-dial enzy mes activities significantly increased and activities of SOD and GSH-Px significantly decreased in nano-TiO2 group,compared with the control group(P <0.05).Cometassay showed that olive tail mo ment (OTM)was significantly increased after nano-TiO2 2 g·kg -1 ,compared with the control group (P<0.05).The histopathology showed ede ma of myocardial cells,myofibril disorders and increasing infla mmatory cells.Vita min C 100,200 and 400 mg·kg -1 can decrease S-T in ECG,OTM,myocardial enzy mes activities,increase the SOD and GSH-Px activities in serum and myocardial tissue;reduce myocardial hypertrophy and infla mmatory cells.CONCLUSION nano-TiO2 can induce myocardial injury inmice and vitamin C can alleviate the da mage.The mechanism may be associated with the antioxidant ability of vitamin C inmyocardial tissue.

Biomarkers in rats for kidney damage characteristics of arsenism due to coal burning and benchmark dose analysis / 中国药理学与毒理学杂志

OBJECTIVE Study the kidney toxic effects caused by burning coal endemic arsenism in rats,application bench mark dose (BMD) method to investigate the bench mark dose of urinary arsenic (UAs)and the changes in bio markers of renal function.METHODS Wistar rats were fed for 90 d with arsenic 0,25,50,100 mg·kg -1 conta minated feed.Urinary arsenic,kidney arsenic and renal function indicators were determined,and routine pathological and fibrosis of kidney were exa mined.UAs as the exposure bio marker,Uβ2-MG,UNAG and UALB for the effect bio markers,application bench mark dose method to calculate the BMD and BMDL of UAs for each effect bio markers.RESULTS UAs,KAs, Uβ2-MG,UNAG,UALB levels of rats in arsenic 100 mg·kg -1 group were increased than normal group (P <0.05);In light microscope,the results of HE staining of rat kidney in all arsenic dose groups showed infla mmatory cell infiltration,renal tubular epithelial cell swelling,renal interstitial capillary dila-tion,congestion and other varying degrees pathological changes,and the results of masson staining showed varying degrees of tubulointerstitial fibrosis;UAs as the exposure bio marker,Uβ2-MG,UNAG, UALB for the effects of mark,the BMD and BMDL of UAs for Uβ2-MG,UNAG,UALB were calculated, the BMD values were 998.9,1213.5,1386.9 μg·g -1 Cr,the BMDL values were 660.5,803.6 and 909. 4 μg·g -1 Cr,respectively.CONCLUSION Burning coal arsenic pollution can cause kidney da mage in rats,mini mal change nephropathy may be the pri mary pathological in the coal arsenic conta mination of kidney da mage.The BMD and BMDL of UAs were 998.9,660.5 μg·g -1 Cr,the early changes of renal function of burning coal arsenism in rats;it is reco mmended to use the more sensitive bio markers Uβ2-MG to calculate the biological exposure li mits on renal injury caused by arsenic.

Study of killing effect on liver cancer cells by transfected dendritic cell vaccine of recombinant plasmid pchIL-18-MAGE in vitro / 中国免疫学杂志

Objective:To observe lethal effect on liver cancer cells by the DCs transfected with recombinant plasmid pchIL -18-MAGE.Methods:Cultivate dendritic cells in vitro ,and recombinant plasmid pchIL-18-MAGE was constructed which was transfected into DCs.Expression of IL-18 and MAGE-1 on the transfected DCs were detected by RT-PCR and Western blot.The diversity of DCs phenotypes after transfect were analyzed by flow cytometry.Different killing effects to the target cells by lymphocytes which were stimulated by DC and those by non-transfected DC were detected.IFN-γin the cellular supernatant of the transfected DCs was inspected by ELISA.Results:pchIL-18-MAGE transfected DCs expressed higher level of CD 83,CD1a,CD86,CD80,HLA-DR,and expressed as a mature DC phenotype.pchIL-18-MAGE group′s killing activity was the highest ( P<0.05 ) .Conclusion: The DC vaccine pchlL-18-MAGE can induce better killing effect compared with other experimental groups.

Detection of Rare Cancer Cells in the Blood by RNA Extraction of Filtered Mononuclear Cells and Reverse Transcription-PCR / 임상검사와정도관리

BACKGROUND: To enhance the cancer cell detection rate in blood, we tried to detect cancer cells by blood filtration, RNA extraction and reverse transcription (RT)-PCR in the filtered mononuclear cells (MNCs). METHODS: From the specimens of whole blood and filtered MNCs, RNA was extracted by the guanidinium isothiocyanate buffer method. Filtration efficiency was evaluated by measurement of leukocyte count, red cell count, and hemoglobin level. To compare the RNA extraction efficiency between whole blood and filtered MNCs, the followings were examined: (1) RNA electrophoresis, (2) hTERT, survivin, plakophilin, LunX, and MAGE A1-6 RT-PCR, and (3) the detection limit of added SNU484 cells in the blood by MAGE A1-6 RT-PCR. Finally blood specimens of 13 lung cancer patients were used to detect cancer cells by LunX and MAGE A1-6 RT-PCR with filtered MNCs. RESULTS: The filtration method revealed 0%, 92.8% and 95.1% filtration rates for leukocyte, red cell, and hemoglobin, respectively. Contamination of concentrated genomic DNA was observed in the electrophoresis of RNA extracted from whole blood. Positive rates of hTERT, survivin and plakophilin were higher in the filtered MNCs. The filtration method detected 1 SNU484 cell/mL, and the blood samples of 4 (30.8%) lung cancer patients showed positive results for RT-PCR. CONCLUSIONS: For the detection of cancer cells in the blood, the filtration method was very efficient, and LunX and MAGE A1-6 genes would be useful for the detection of blood cancer cells in patients with lung cancer.

A novel DNA vaccine constructed by heat shock protein 70 and melanoma antigen-encoding gene 3 against tumorigenesis.

Melanoma antigen-encoding gene 3 (MAGE-3) is an ideal candidate for a tumor vaccine although its potency need to be increased. Heat shock proteins (HSPs) represents a potential approach for increasing the potency of DNA vaccines. In the present study, a fusion DNA vaccine composed of Mycobacterium tuberculosis HSP70 and MAGE-3 was constructed and used to immunize C57BL/6 mice against B16 or B16-MAGE-3 tumor cells. The results show that the HSP70-MAGE-3 fusion DNA vaccine enhanced the frequency of MAGE-3-specific cytotoxic T-cells as compared to the MAGE-3 DNA vaccine or the HSP70/MAGE-3 cocktail DNA vaccine (P<0.05). In conclusion, the results indicate that the HSP70-MAGE-3 fusion DNA vaccine can strongly activate MAGE-3 specific cellular immunological reactions and thus significantly inhibit the growth of B16-MAGE-3 tumors, improving the survival of tumor-bearing mice, and the HSP70-MAGE-3 fusion DNA vaccine has a significant therapeutic effect on the tumors that express MAGE-3 antigens.

Identification and Immune response of Murine MAGE-3 Derived MHC-Ⅰ/MHC-ⅡRestricted Peptide Epitope / 中山大学学报(医学科学版)

[Objective] To design routine MAGE-3 derived MHC-I/MHC-II restricted peptide epitope, which containing CD4~+-CD8~+ T cell epitope peptides antigen. [Methods] The epitope peptides were made through computer simulation designing, and peptide epitopes qualification tests were performed after the synthesis of peptide antigen, ELISPOT and cell-toxic analysis were used to evaluate the proliferation ability and cytokine-release ability of peptide-stimulated T cell. [Results] The sequence of obtained MAGE-3 derived restricted epitope peptide was FITC-YEEYYPLIFLDNDQETMETSEEEEYEEYYPLIF, of which the purity ≥ 90% tested by high performance liquid chromatography. MAGE-3 epitope peptide antigen could induce T lymphocyte proliferation, and induce T lymphocyte to secret IFN-γ, which higher than that of the control group (49 vs. 6 spots/10~6, P≤0.05 ). MAGE-3 epitope peptide could induce cytotoxic T lymphocytes to cause 42% of MFC cell lysis rupture, higher than control group (P≤0.05). [Conclusion] CD4~+-CD8~+ T cell epitope MAGE-3 peptide antigen showed considerable immunological effect in vitro, and such a peptide antigen can work as therapeutic polypeptide vaccine for H-2K~K mice gastric cancer which express MAGE-3 antigen.

Melanoma antigen-A family in tumor / 国际肿瘤学杂志

As a tumor-specific antigen highly expressed in various types of tumors, MACE-A does not exist in normal adult tissues, except for testis and placenta. Therefore MAGE-A antigens are regarded. tumor specific antigen,and have significant significance for cancer immunotherapy.

Use of Dentritic Cells Pulsed with HLA-A2-restricted MAGE-A1 Peptide to Generate Cytotoxic T Lymphocytes against Malignant Glioma / 华中科技大学学报（医学）（英德文版）

This study developed a novel approach of targeting malignant glioma with pMAGE-A1278-286-specific cytotoxic T lymphocytes (CTLs) induced from the peripheral blood mononuclear cells of healthy donors by multiple stimulations with human leukocyte antigen (HLA)-A2-restricted pMAGE-A1278-286 peptide-pulsed dentritic cells. Cytotoxic assays were performed by the colorimetric CytoTox 96 assay to analyze cytotoxic activity of the induced CTLs against various target cells. The induced CTLs showed approximately 45% specific lysis against T2pMAGE-A 1278-286 (pMAGE-A 1278-286 peptide pulsed T2 cells) and U251 (HLA-A2+, MAGE-A 1 +)at an effector:target ratio of 40:1, and approximately 5% cytolysis against T2pHIV, A172 (HLA-A2,MAGE-AI+), K562 and T2 cells without being pulsed with peptide at any effector:target ratio. The specific killing activity of the induced CTLs against T2pMAGE-A1278-286 and U251 was much more obvious than in any other control group (P＜0.05). The cytotoxic activity against the T2pMAGE-A1278-286 and U251 was significantly eliminated by anti-HLA class I mAb W6/32.These results suggest that pMAGE-A1278-286 epitope may serve as a surrogate tumor antigen target of specific immunotherapy for treating HLA-A2 patients with malignant glioma.