Monoclonal Antibodies to Esophageal Cancer–Related Gene2 Protein

Huang, Ge; Wang, Debin; Guo, Liping; Zhao, Ning; Li, Yuanyuan; Lu, Shih Hsin

doi:10.1089/hyb.2005.24.86

Cited by 9 publications

(5 citation statements)

References 17 publications

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“…Forty‐eight hours after transient transfection, cell lysates were acquired and Western blot was performed as described previously using mouse anti‐ ECRG2 monoclonal antibody generated by our laboratory [19]. The results were scanned by LAS4000 imaging system (Fuji Film, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

MicroRNA‐1322 regulates ECRG2 allele specifically and acts as a potential biomarker in patients with esophageal squamous cell carcinoma

Zhang

Zhao

Wang

et al. 2012

Molecular Carcinogenesis

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A short tandem repeat (STR) polymorphism in the 3'UTR region of esophageal cancer-related gene 2 (ECRG2, also known as SPINK7) has been widely reported to be associated with the incidence and the prognosis of esophageal squamous cell carcinoma (ESCC). This study explores how the microRNA binding to the STR region affects ECRG2 expression in ESCC. Dual-luciferase reporter assays were used to verify the effects of the four microRNAs (miR-580, miR-1182, miR-1272, and miR-1322) predicted to bind the STR region of the ECRG2 3' untranslated region (UTR). The expression of identified effective microRNA was then analyzed in 44 paired ESCC and adjacent normal tissues and 402 case-controlled serum samples (divided into a discovery group and an independent validation group) by real-time RT-PCR assay. We found that only miR-1322 could significantly down-regulate the ECRG2 with TCA3 allele (P < 0.01), but it could not down-regulate the ECRG2 with TCA4 allele significantly (P > 0.05). MiR-1322 was also expressed significantly higher in ESCC tissue and serum samples than in controls (both P < 0.01). Additionally, serum levels of miR-1322 yielded an under receiver operating characteristic (ROC) curve area of 0.847 (95% CI, 0.795-0.890) for discriminating ESCCs from healthy controls in the discovery group and a similar result was obtained in the validation group (under ROC area is 0.845; 95%CI, 0.780-0.897). We conclude that miR-1322 can regulate ECRG2 in an allele-specific manner and that serum levels of miR-1322 can serve as a potential diagnostic biomarker for patients with ESCC.

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Section: Methodsmentioning

confidence: 99%

MicroRNA‐1322 regulates ECRG2 allele specifically and acts as a potential biomarker in patients with esophageal squamous cell carcinoma

Zhang

Zhao

Wang

et al. 2012

Molecular Carcinogenesis

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“…Reagents-Monoclonal anti-ECRG2 antibody was produced by our laboratory (22,23). Human uPA and uPAR monoclonal antibodies were purchased from American Diagnostica (Greenwich, CT).…”

Section: Methodsmentioning

confidence: 99%

ECRG2 Regulates Cell Migration/Invasion through Urokinase-type Plasmin Activator Receptor (uPAR)/β1 Integrin Pathway

Cheng

Shen

Yin

et al. 2009

Journal of Biological Chemistry

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ECRG2 is a novel gene that shows sequence similarity to KAZAL-type serine protease inhibitor. We have previously demonstrated that ECRG2 inhibits migration/invasion of lung cancer PG cells. However, the mechanism by which ECRG2 performs these activities is a compelling question. Urokinase-type plasmin activator (uPA) binding to uPAR induces migration/ invasion through multiple interactors including integrins. In this study, we found that ECRG2 binds specifically to the kringle domain of uPA. Moreover, we demonstrated that ECRG2 forms a complex with uPA⅐uPAR, that such a complex modifies the dynamical association of uPAR with ␤1 integrins, and that disruption inhibits Src/MAP (mitogen-activated protein) kinase pathway, resulting in suppression of cell migration/invasion in an in vitro Matrigel migration/invasion assay. Conversely, depletion of ECRG2 markedly enhanced the association of uPAR with ␤1 integrins, elevated basal Src/MAP kinase activation, and stimulated HT1080, MDA-MB-231, and MCF-7 cell migration/invasion. Together, our results provide evidence that ECRG2 is involved in the regulation of migration/invasion through uPA/uPAR/␤1 integrins/Src/MAP kinase pathway and may represent a novel therapeutic target for cancer.Tumor cells must invade through the adjacent basement membrane into surrounding tissues and then migrate to and invade the vasculature to metastasize to distant sites (1). The processes of tumor cell migration and invasion involve a dynamic interaction between the tumor cells and the extracellular matrix and are regulated by multiple cytokines and growth factors, integrins, cell-cell adhesion molecules/communication, matrix-degrading enzymes,

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“…Generation of ECRG2 Antibody-We generated anti-ECRG2 monoclonal antibodies by using the C-terminal of ECRG2 peptide (KSNGRVQFLHDGSC) as immunogen (28). 217-258 encoding portions of human ECRG2 were amplified using Pfu polymerase (Stratagene), then cloned into pGEX-4T-3 (GE Healthcare) as BamHI/NotI restriction fragments.…”

Section: Methodsmentioning

confidence: 99%

“…Glutathione S-transferase fusion proteins were expressed in BL21 Escherichia coli cells by induction with 1 mM isopropyl 1-thio-␤-Dgalactopyranoside at 37°C then purified using glutathioneSepharose beads (GE Healthcare) according to the manufacturer's instructions. The anti-ECRG2 monoclonal antibody was generated following procedures described previously (28). Briefly, mice were immunized with glutathione S-transferase-ECRG2.…”

Section: Methodsmentioning

confidence: 99%

ECRG2 Disruption Leads to Centrosome Amplification and Spindle Checkpoint Defects Contributing Chromosome Instability

Cheng

Shen

Yang

et al. 2008

Journal of Biological Chemistry

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Cancer cells contain an abnormal number of chromosomes (aneuploidy), which is a prevalent form of genetic instability in human cancers. Abnormal amplification of centrosomes and defects of spindle assembly checkpoint are the major causes of chromosome instability in cancer cells. Here we present biochemical evidence to suggest a role of ECRG2, a novel tumor suppressor gene, in maintaining chromosome stability. ECRG2 localized to centrosomes during interphase and kinetochores during mitosis. Further analysis revealed that ECRG2 participates in centrosome amplification in a p53-dependent manner. Depletion of ECRG2 not only destabilized p53, down-regulated p21, and increased the cyclin E/CDK2 activity, thus initiating centrosome amplification, but also abolished the ability of p53 localize to centrosomes. Overexpression of ECRG2 restored the p53-dependent suppression of centrosome duplication. Furthermore, ECRG2-depleted cells show severely disrupted spindle phenotype but fail to maintain the mitotic arrest due to minimal BUBR1 protein levels. Taken together, our results indicate that ECRG2 is important for ensuring centrosome duplication, spindle assembly checkpoint, and accurate chromosome segregation, and its depletion may contribute to chromosome instability and aneuploidy in human cancers.

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Monoclonal Antibodies to Esophageal Cancer–Related Gene2 Protein

Cited by 9 publications

References 17 publications

MicroRNA‐1322 regulates ECRG2 allele specifically and acts as a potential biomarker in patients with esophageal squamous cell carcinoma

MicroRNA‐1322 regulates ECRG2 allele specifically and acts as a potential biomarker in patients with esophageal squamous cell carcinoma

ECRG2 Regulates Cell Migration/Invasion through Urokinase-type Plasmin Activator Receptor (uPAR)/β1 Integrin Pathway

ECRG2 Disruption Leads to Centrosome Amplification and Spindle Checkpoint Defects Contributing Chromosome Instability

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