AIM: To validate the utility of Annexin A10 as a surrogate marker of the serrated neoplasia pathway in invasive colorectal cancers(CRCs).METHODS: A total of 1133 primary CRC patients who underwent surgical resection a...AIM: To validate the utility of Annexin A10 as a surrogate marker of the serrated neoplasia pathway in invasive colorectal cancers(CRCs).METHODS: A total of 1133 primary CRC patients who underwent surgical resection at Seoul National University Hospital between January 2004 and December 2007 were enrolled.Expression of Annexin A10 was evaluated by immunohistochemistry using tissue microarray and paired to our findings on clinicopathologic and molecular characteristics of each individual.Cp G island methylator phenotype was determined by Methy Light assay and microsatellite instability was determined by high performance liquid chromatography.KRAS and BRAF mutation status was evaluated by direct sequencing and allele-specific PCR.Univariate and stage-specific survival analyses were performed to reveal the prognostic value of Annexin A10 expression.RESULTS: Annexin A10 expression was observed in 66(5.8%) of the 1133 patients.Annexin A10 expression was more commonly found in females and was associated with proximal location,ulcerative gross type,advanced T category,N category and TNM stage.CRCs with Annexin A10 expression showed an absence of luminal necrosis,luminal serration and mucin production.CRCs with Annexin A10 expression were associated with Cp G island methylator phenotype,microsatellite instability and BRAF mutation.In survival analysis,Annexin A10 expression was associated with poor overall survival and progression-free survival,especially in stage Ⅳ CRCs.CONCLUSION: Annexin A10 expression is associated with poor clinical behavior and can be used a supportive surrogate marker of the serrated neoplasia pathway in invasive CRCs.展开更多
Aberrant expression of annexin A2-SI00A10 heterotetramer （AIIt） associated with PML/RARα fusion protein causes lethal hyperfibrinolysis in acute promyelocytic leukemia （APL）, but the mechanism is unclear. To faci...Aberrant expression of annexin A2-SI00A10 heterotetramer （AIIt） associated with PML/RARα fusion protein causes lethal hyperfibrinolysis in acute promyelocytic leukemia （APL）, but the mechanism is unclear. To facilitate the investigation of regulatory association between ANXA2 and promyelocytic leukemia/retinoic acid receptor α （PML/RARα） fusion protein, this work was performed to determine the transcription start site of ANXA2 promoter with rapid amplification of 5＇-cDNA ends analysis. Zinc-induced U937/PR9 cells expressed PML/RARα fusion protein, and resultant increases in ANXA2 transcripts and translational expressions of both ANXA2 and S100A10, while S100A10 transcripts remained constitutive. The transactivation of ANXA2 promoter by PML/RARα fusion protein was 3.29 ± 0.13 fold higher than that by control pSG5 vector or wild-type RARer. The overexpression of ANXA2 in U937 transfected with full-length ANXA2 cDNA was associated with increased S100A10 subunit, although S100A10 transcripts remained constitutive. The tPA-dependent initial rate of plasmin generation （IRPG） in zinc-treated U937/PR9 increased by 2.13-fold, and cell invasiveness increased by 27.6%. Antibodies against ANXA2, S100A10, or combination of both all remarkably inhibited the IRPG and invasiveness in U937/PR9 and NB4. Treatment of zinc-induced U937/PR9 or circulating APL blasts with all-trans retinoic acid （ATRA） significantly reduced cell surface ANXA2 and S100A10 and associated reductions in IRPG and invasiveness. Thus, PML/RARα fusion protein transactivated the ANXA2 promoter to upregulate ANXA2 and accumulate S100A10. Increased AIIt promoted IRPG and invasiveness, both of which were partly abolished by antibodies against ANXA2 and S100A10 or by ATRA.展开更多
Backgroud in tumors the process of apoptosis occurs over an interval of time after chemotherapy, it is important to determine the best time for detecting apoptosis by in vivo imaging. In this study, we evaluated the d...Backgroud in tumors the process of apoptosis occurs over an interval of time after chemotherapy, it is important to determine the best time for detecting apoptosis by in vivo imaging. In this study, we evaluated the dynamics and feasibility of imaging non-small cell lung cancer （NSCLC） apoptosis induced by paclitaxel treatment using a 99Tcm-labeled Annexin V recombinant with ten consecutive histidines （Hislo-Annexin V） in a mouse model. Methods 99Tcm-His10-Annexin V was prepared by one step direct labeling; radio-chemical purity （RCP） and radio-stability was tested. The binding of 99Tcm-His10-Annexin V to apoptotic cells was validated in vitro using camptothecin-induced Jurkat cells. In vivo bio-distribution was determined in mice by dissection. The human H460 NSCLC tumor cell line （H460） tumor-bearing mice were treated with intravenous paclitaxel 24, 48 and 72 hours later. 99Tcm-His10-Annexin V was injected intravenously, and planar images were acquired at 2, 4 and 6 hours post-injection on a dual-head gamma camera fitted with a pinhole collimator. Tumor-to-normal tissue ratios （T/NT） were calculated by ROI analysis and they reflected specific binding of 99Tcm-His10-Annexin V. Mice were sacrificed after imaging. Caspase-3, as the apoptosis detector, was determined by flow cytometry, and DNA fragmentation was analyzed by the terminal deoxynucleotidytransferase mediated dUTP nick-end labeling （TUNEL） assay. Nonspecific accumulation of protein was estimated using bovine serum albumin （BSA）. The imaging data were correlated with TUNEL-positive nuclei and caspase-3 activity. Results 99Tcm-His10-Annexin V had a RCP 〉98% and high stability 2 hours after radio-labeling, and it could bind to apoptotic cells with high affinity. Bio-distribution of 99Tcm-His10-Annexin V showed predominant uptake in kidney, relatively low uptake in myocardium, liver and gastrointestinal tract, and rapid clearance from blood and kidney was observed. The TINT was significantly increased after paclitaxel treatment展开更多
基金Supported by Grant from Basic Science Research Program through the National Research Foundation(NRF)funded by the Ministry of Education,No.2013R1A1A2059080a grant from the Korean Health Technology R&D Project,Ministry of Health&Welfare,No.HI13C1804+2 种基金Priority Research Centers Program through the NRF funded by the Ministry of Education,Science and Technology,No.2009-0093820the NRF grant funded by the Ministry of Science,ICT,and Future planning,No.2011-0030049a grant of the Korea Health Technology R&D Project,Ministry of Health&Welfare,Republic of Korea,No.HI14C1277."
文摘AIM: To validate the utility of Annexin A10 as a surrogate marker of the serrated neoplasia pathway in invasive colorectal cancers(CRCs).METHODS: A total of 1133 primary CRC patients who underwent surgical resection at Seoul National University Hospital between January 2004 and December 2007 were enrolled.Expression of Annexin A10 was evaluated by immunohistochemistry using tissue microarray and paired to our findings on clinicopathologic and molecular characteristics of each individual.Cp G island methylator phenotype was determined by Methy Light assay and microsatellite instability was determined by high performance liquid chromatography.KRAS and BRAF mutation status was evaluated by direct sequencing and allele-specific PCR.Univariate and stage-specific survival analyses were performed to reveal the prognostic value of Annexin A10 expression.RESULTS: Annexin A10 expression was observed in 66(5.8%) of the 1133 patients.Annexin A10 expression was more commonly found in females and was associated with proximal location,ulcerative gross type,advanced T category,N category and TNM stage.CRCs with Annexin A10 expression showed an absence of luminal necrosis,luminal serration and mucin production.CRCs with Annexin A10 expression were associated with Cp G island methylator phenotype,microsatellite instability and BRAF mutation.In survival analysis,Annexin A10 expression was associated with poor overall survival and progression-free survival,especially in stage Ⅳ CRCs.CONCLUSION: Annexin A10 expression is associated with poor clinical behavior and can be used a supportive surrogate marker of the serrated neoplasia pathway in invasive CRCs.
文摘Aberrant expression of annexin A2-SI00A10 heterotetramer （AIIt） associated with PML/RARα fusion protein causes lethal hyperfibrinolysis in acute promyelocytic leukemia （APL）, but the mechanism is unclear. To facilitate the investigation of regulatory association between ANXA2 and promyelocytic leukemia/retinoic acid receptor α （PML/RARα） fusion protein, this work was performed to determine the transcription start site of ANXA2 promoter with rapid amplification of 5＇-cDNA ends analysis. Zinc-induced U937/PR9 cells expressed PML/RARα fusion protein, and resultant increases in ANXA2 transcripts and translational expressions of both ANXA2 and S100A10, while S100A10 transcripts remained constitutive. The transactivation of ANXA2 promoter by PML/RARα fusion protein was 3.29 ± 0.13 fold higher than that by control pSG5 vector or wild-type RARer. The overexpression of ANXA2 in U937 transfected with full-length ANXA2 cDNA was associated with increased S100A10 subunit, although S100A10 transcripts remained constitutive. The tPA-dependent initial rate of plasmin generation （IRPG） in zinc-treated U937/PR9 increased by 2.13-fold, and cell invasiveness increased by 27.6%. Antibodies against ANXA2, S100A10, or combination of both all remarkably inhibited the IRPG and invasiveness in U937/PR9 and NB4. Treatment of zinc-induced U937/PR9 or circulating APL blasts with all-trans retinoic acid （ATRA） significantly reduced cell surface ANXA2 and S100A10 and associated reductions in IRPG and invasiveness. Thus, PML/RARα fusion protein transactivated the ANXA2 promoter to upregulate ANXA2 and accumulate S100A10. Increased AIIt promoted IRPG and invasiveness, both of which were partly abolished by antibodies against ANXA2 and S100A10 or by ATRA.
文摘Backgroud in tumors the process of apoptosis occurs over an interval of time after chemotherapy, it is important to determine the best time for detecting apoptosis by in vivo imaging. In this study, we evaluated the dynamics and feasibility of imaging non-small cell lung cancer （NSCLC） apoptosis induced by paclitaxel treatment using a 99Tcm-labeled Annexin V recombinant with ten consecutive histidines （Hislo-Annexin V） in a mouse model. Methods 99Tcm-His10-Annexin V was prepared by one step direct labeling; radio-chemical purity （RCP） and radio-stability was tested. The binding of 99Tcm-His10-Annexin V to apoptotic cells was validated in vitro using camptothecin-induced Jurkat cells. In vivo bio-distribution was determined in mice by dissection. The human H460 NSCLC tumor cell line （H460） tumor-bearing mice were treated with intravenous paclitaxel 24, 48 and 72 hours later. 99Tcm-His10-Annexin V was injected intravenously, and planar images were acquired at 2, 4 and 6 hours post-injection on a dual-head gamma camera fitted with a pinhole collimator. Tumor-to-normal tissue ratios （T/NT） were calculated by ROI analysis and they reflected specific binding of 99Tcm-His10-Annexin V. Mice were sacrificed after imaging. Caspase-3, as the apoptosis detector, was determined by flow cytometry, and DNA fragmentation was analyzed by the terminal deoxynucleotidytransferase mediated dUTP nick-end labeling （TUNEL） assay. Nonspecific accumulation of protein was estimated using bovine serum albumin （BSA）. The imaging data were correlated with TUNEL-positive nuclei and caspase-3 activity. Results 99Tcm-His10-Annexin V had a RCP 〉98% and high stability 2 hours after radio-labeling, and it could bind to apoptotic cells with high affinity. Bio-distribution of 99Tcm-His10-Annexin V showed predominant uptake in kidney, relatively low uptake in myocardium, liver and gastrointestinal tract, and rapid clearance from blood and kidney was observed. The TINT was significantly increased after paclitaxel treatment
