Journal of capillary electrophoresis最新文献

Proteomic analysis of spheroids of rhabdomyosarcoma cells cultured with decellularized muscle extracts 脱细胞肌肉提取物培养横纹肌肉瘤细胞球体的蛋白质组学分析

Journal of capillary electrophoresis Pub Date : 2022-01-01 DOI: 10.2198/jelectroph.66.1

Yooksil Sin, T. Ono, R. Tsuchiya, Rei Noguchi, Yuki Yoshimatsu, H. Kosako, T. Kondo

引用次数: 0

Use of Escherichia coli expression system for analyzing kinase motifs 利用大肠杆菌表达系统分析激酶基序

Journal of capillary electrophoresis Pub Date : 2022-01-01 DOI: 10.2198/jelectroph.66.13

Emiko Kinoshita-Kikuta, Yoko Ino, Y. Kimura, Tomoko Akiyama, E. Kinoshita, T. Koike

引用次数: 0

Electrophoretic extraction of protein complexes after separation and detection by a combined method of non-denaturing two-dimensional electrophoresis and reversible staining 电泳提取分离和检测后的蛋白质复合物，采用非变性双向电泳和可逆染色相结合的方法

Journal of capillary electrophoresis Pub Date : 2022-01-01 DOI: 10.2198/jelectroph.66.5

Karin Nakao, Y. Shimazaki

引用次数: 1

Proteogenomic approach to drug targets in osteosarcomas with different original sites 不同原发部位骨肉瘤药物靶点的蛋白质基因组学研究

Journal of capillary electrophoresis Pub Date : 2021-01-01 DOI: 10.2198/JELECTROPH.65.1

Rei Noguchi, Yuki Yoshimatsu, T. Ono, Akane Sei, T. Kondo

{"title":"Proteogenomic approach to drug targets in osteosarcomas with different original sites","authors":"Rei Noguchi, Yuki Yoshimatsu, T. Ono, Akane Sei, T. Kondo","doi":"10.2198/JELECTROPH.65.1","DOIUrl":"https://doi.org/10.2198/JELECTROPH.65.1","url":null,"abstract":"Regulation of kinase activity plays a crucial role in carcinogenesis and cancer progression. Mutations in the activity domain of kinases are extensively investigated as therapeutic targets. We examined anti-proliferative anti-cancer drugs and drug targets via the multi-omics approach: (i) comprehensive kinase activity assay, (ii) high-throughput drug screening, and (iii) genomic sequencing. Two osteosarcomas cell lines, NCC-OS1-C1 and NCC-ESOS1-C1 derived from bone and soft tissue respectively, were used. Genetic alterations were examined by NCC Oncopanel based on the next-generation sequencing technology and SNP array. One hundred kinases were monitored by the PamStation 12, an in vitro kinase assay. The anti-proliferative effects of 214 FDA-approved anti-cancer drugs were examined. Mutation of PIK3CA and deletion of CDKN2A were identified in NCC-ESOS1-C1 and druggable genetic alterations were not identified in the NCC-OS1-C1. PI3K-AKT pathway or CDKN2A inhibitors did not show significant effects on these cell lines. Comprehensive kinomic assay revealed no remarkable differences on these osteosarcoma cells (R2=0.99). The two cells shared similar kinase activity profiles for FES, FER, PDGFR-β, VEGFR2, and Wee1. Anti-proliferative effects of anti-cancer drugs on NCC-OS1-C1 and NCC-ESOS1 cells showed remarkable differences. Significant responses to romidepsin and trabectedin were observed for both. Eribulin was effective on NCCOS1-C1; ifosfamide and dacarbazine were effective on NCC-ESOS1-C1 only. Hence, investigating kinase activities and genetic alterations will lead to predict the effects of kinase inhibitors. The different status of kinase mutations, activities, and response to inhibitors should be integrated. Multi-omics experiments and data integration are crucial in understanding cancer progression and developing novel therapies.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90419237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Comprehensive miRNA expression analysis for histological subtypes of soft tissue sarcoma 软组织肉瘤组织学亚型miRNA综合表达分析

Journal of capillary electrophoresis Pub Date : 2021-01-01 DOI: 10.2198/JELECTROPH.65.13

R. Tsuchiya, Yuki Yoshimatsu, Naoto Tsuchiya, S. Ohtori, A. Kawai, T. Kondo

{"title":"Comprehensive miRNA expression analysis for histological subtypes of soft tissue sarcoma","authors":"R. Tsuchiya, Yuki Yoshimatsu, Naoto Tsuchiya, S. Ohtori, A. Kawai, T. Kondo","doi":"10.2198/JELECTROPH.65.13","DOIUrl":"https://doi.org/10.2198/JELECTROPH.65.13","url":null,"abstract":"Sarcoma is a rare mesenchymal malignancy that comprises more than 50 histological subtypes. Because of the rarity and diversity of sarcomas, their differential diagnosis is difficult, and there is still a need for biomarkers to support pathological diagnoses. Micro RNAs (miRNAs) are small noncoding RNAs that regulate the behavior of tumors, such as invasion and metastasis. The expression patterns of miRNAs reflect the origin of malignancy and are considered to be candidate biomarkers. To understand the molecular background of those histological subtypes, we investigated the miRNA expression in 89 tumor tissues of eight subtypes. The correlation coefficients between each sarcoma subtype on the basis of miRNA expression values were mostly higher than 0.7, reflecting the common mesenchymal origin. By contrast, hierarchical clustering and principal component analysis showed that three types of sarcoma with chromosomal translocation (i.e., dermatofibrosarcoma protuberans, myxoid liposarcoma, and synovial sarcoma) were grouped according to their histological subtypes, whereas five types with complex karyotypes (i.e., myxofibrosarcoma, malignant peripheral nerve sheath tumor, undifferentiated pleomorphic sarcoma, dedifferentiated liposarcoma, and pleomorphic liposarcoma) were not. Notably, the number of miRNAs whose expression pattern was unique to histological subtypes with statistical significance was higher in sarcomas with chromosome translocation than in those with complex karyotypes. Hence, it can be concluded that the miRNAs unique to histological subtypes are candidate biomarkers for the differential diagnosis of sarcomas, particularly in those with chromosomal translocation.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"122 17","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91403052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-based biotyping of silk. I. Method development 基于基质辅助激光解吸/电离飞行时间质谱的蚕丝生物分型。一、方法开发

Journal of capillary electrophoresis Pub Date : 2021-01-01 DOI: 10.2198/jelectroph.65.39

H. Kajiwara, R. Murakami, K. Nakajima

引用次数: 0

Drug screening and kinase activity profiling of a novel patient-derived cell line of clear cell ovarian carcinoma 一种新型透明细胞卵巢癌患者来源细胞系的药物筛选和激酶活性分析

Journal of capillary electrophoresis Pub Date : 2021-01-01 DOI: 10.2198/jelectroph.65.23

Rei Noguchi, Yuki Yoshimatsu, Akane Sei, H. Yoshida, Tomoyasu Katou, T. Kondo

{"title":"Drug screening and kinase activity profiling of a novel patient-derived cell line of clear cell ovarian carcinoma","authors":"Rei Noguchi, Yuki Yoshimatsu, Akane Sei, H. Yoshida, Tomoyasu Katou, T. Kondo","doi":"10.2198/jelectroph.65.23","DOIUrl":"https://doi.org/10.2198/jelectroph.65.23","url":null,"abstract":"Clear cell carcinoma (CCC) is a rare subtype of ovarian cancer resistant to standard platinum chemotherapy, which leads to a poor prognosis for patients with CCC. Kinases are targets for anticancer drugs; few studies have profiled kinase activity to identify kinase inhibitors as novel anticancer drugs. In this study, we aimed to identify novel anticancer drugs for the treatment of CCC with comprehensive kinase activity assay and drug screening. Using ascites from a 51-year old patient, we established and characterized the NCC-cOV1-C1 cell line. We screened the antiproliferative effects of 152 small anticancer compounds and conducted comprehensive kinase activity assays with the PamStation12 platform. The NCC-cOV1-C1 cells harbor copy number variation of HFN1β amplification, and exhibit constant growth, spheroid formation, and invasion capability. NCC-cOV1-C1 cells responded remarkably to idarubicin HCl and vorinostat. The kinase activity assay revealed that SRC and EGFR were highly activated in NCC-cOV1-C1 cells; the SRC inhibitor dasatinib and the EGFR inhibitor lapatinib exhibited antiproliferative effects and down-regulation of downstream signaling. The NCC-cOV1-C1 cell line will be a useful tool for basic and preclinical study of CCC, and the clinical utility of idarubicin HCl, vorinostat, dasatinib, lapatinib is worthy of further investigation.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83538150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Gel electrophoresis for phosphorylated proteins: a brief introduction 磷酸化蛋白凝胶电泳:简要介绍

Journal of capillary electrophoresis Pub Date : 2020-01-01 DOI: 10.2198/jelectroph.64.13

Rei Noguchi, Yooksil Sin, T. Kondo

{"title":"Gel electrophoresis for phosphorylated proteins: a brief introduction","authors":"Rei Noguchi, Yooksil Sin, T. Kondo","doi":"10.2198/jelectroph.64.13","DOIUrl":"https://doi.org/10.2198/jelectroph.64.13","url":null,"abstract":"SUMMARY Protein phosphorylation is a key mechanism that regulates cellular physiological functions such as proliferation, migration, cell cycle progression, apoptosis, and differentiation. Aberrations in kinase activity and subsequent dysregulation of protein phosphorylation occur in the process of carcinogenesis and cancer progression and are considered to be therapeutic targets and biomarkers in oncology. Gel electrophoresis has versatile utility in the study of protein phosphorylation. Phosphorylated proteins can be enriched prior to gel electrophoresis, and the proteins separated by gel electrophoresis are visualized by colorimetric methods or western blotting with antibodies, which specifically detect phosphorylation. Phosphorylated proteins migrate differently from non-phosphorylated proteins in gels containing substrates with affinity for phosphorylation. All these methods can be combined in multiple ways, generating unique data from different viewpoints. The identification of separated proteins can be achieved by mass spectrometry, making it possible to integrate protein and genetic data. Peptide array allow the evaluation of kinase activity in heterogeneous samples. As protein phosphorylation and kinase activity are under the regulation of multiple mechanisms and their statuses influence the structures and functions of the other proteins, a multidisciplinary approach is required, and gel electrophoresis will play an important role in the study of protein phosphorylation.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86985357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A dot-blot-staining method for detecting phosphoproteins with a Phos-tag Aqua fluorescent dye 一种用Phos-tag Aqua荧光染料检测磷蛋白的点印迹染色方法

Journal of capillary electrophoresis Pub Date : 2020-01-01 DOI: 10.2198/jelectroph.64.7

Emiko Kinoshita-Kikuta, K. Akayama, E. Kinoshita, T. Koike

引用次数: 0

Production of dye-binding esterase reactor after separation and detection using combined native isoelectric focusing and blue native electrophoresis 采用天然等电聚焦和蓝色天然电泳相结合分离检测后的染料结合酯酶反应器的生产

Journal of capillary electrophoresis Pub Date : 2020-01-01 DOI: 10.2198/jelectroph.64.1

Y. Shimazaki, Hikari Nakamaru

引用次数: 1