Acquisition of useful sero-diagnostic autoantibodies using the same patients’sera and tumor tissues

Kobayashi, Makoto; Nagashio, Ryo; Ryuge, Shinichiro; Murakami, Yousuke; Yanagita, Kengo; Nakashima, Hiroyasu; Matsumoto, Toshihide; Jiang, Shi Xu; Saegusa, Makoto; Satoh, Yukitoshi; Masuda, Noriyuki; Sato, Yuichi

doi:10.2220/biomedres.35.133

Cited by 8 publications

(4 citation statements)

References 25 publications

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“…Sample preparation for this analysis was described previously (Kobayashi et al, 2014) and tricine 2-DE was performed with minor modification. The first-dimensional isoelectric focusing gel was produced with carrier ampholyte at pH 3-10 (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA).…”

Section: Tricine Two-dimensional Gel Electrophoresis and Protein Identificationmentioning

confidence: 99%

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S100A16 is a Prognostic Marker for Lung Adenocarcinomas

Saito¹,

Kobayashi²,

Nagashio³

et al. 2015

Asian Pacific Journal of Cancer Prevention

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Background: Many functional molecules controlling diverse cellular function are included in low-molecular weight proteins and peptides. Materials and Methods: To identify proteins controlling function in lung adenocarcinomas (AC), we performed two-dimensional gel electrophoresis employing tricine-SDS polyacrylamide in the second dimension (tricine 2-DE). This system was able to detect proteins under 1 kDa even with posttranslational modifications. To confirm the utility of detected proteins as novel tumor markers for AC, we performed immunohistochemical analysis using 170 formalin-fixed and paraffin-embedded lung AC tissues. Results: Tricine 2-DE revealed that five proteins including S100A16 were overexpressed in lung AC-derived cells compared with lung squamous cell carcinoma, small cell carcinoma, and large cell neuroendocrine carcinomaderived cells. Immunohistochemically, S100A16 showed various subcellular localization in lung cancer tissues and a membranous staining status was correlated with the T-factor (P=0.0008), pathological stage (P=0.0015), differentiation extent (P=0.0001), lymphatic invasion (P=0.0007), vascular invasion (P=0.0001), pleural invasion (P=0.0087), and gender (P=0.039), but not with the age or smoking history. More importantly, membranous staining of S100A16 was significantly correlated with a poorer overall survival of either stage I (P=0.0088) or stage II / III (P=0.0003) lung AC patients, and multivariate analysis confirmed that membranous expression of S100A16 was an independent adverse prognostic indicator (P=0.0001). Conclusions: The present results suggest that S100A16 protein is a novel prognostic marker for lung AC.

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Section: Tricine Two-dimensional Gel Electrophoresis and Protein Identificationmentioning

confidence: 99%

“…Protein spots that changed more than 1.5-fold between AC cells and other cells progressed to in-gel digestion. In-gel digestion and protein identification were conducted as previously described (Kobayashi et al, 2014).…”

Section: Tricine Two-dimensional Gel Electrophoresis and Protein Identificationmentioning

confidence: 99%

S100A16 is a Prognostic Marker for Lung Adenocarcinomas

Saito¹,

Kobayashi²,

Nagashio³

et al. 2015

Asian Pacific Journal of Cancer Prevention

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“…The generation of monoclonal antibodies was previously described [3] [4]. The antigen of one antibody designated as Ku-Lad-001 was identified by immunoprecipitation (IP) and MALDI-TOF/TOF MS analysis and characterized by immunoblotting (IB) and immunohistochemistry (IHC) [6]. Recombinant CANX protein and Venus protein as negative controls with FLAG-tag were prepared using a wheat germ cell-free system [7].…”

Section: Generation Of Monoclonal Antibodies and Antigen Identificatimentioning

confidence: 99%

Calnexin is a novel sero-diagnostic marker for lung cancer

Kobayashi¹,

Nagashio²,

Jiang³

et al. 2015

Lung Cancer

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To develop sero-diagnostic markers for lung cancer, we generated monoclonal antibodies using lung adenocarcinoma (AC)-derived A549 cells as antigens by employing the random immunization method. Hybridoma supernatants were immunohistochemically screened for antibodies with AMeX-fixed and paraffin-embedded A549 cell preparations. Positive clones were monocloned twice through limiting dilutions. From the obtained monoclonal antibodies, one designated as KU-Lad-001 was recognized as calnexin (CANX) based on immunoprecipitation and MADLI TOF/TOF-MS analysis. To evaluate the utility of this antibody as a sero-diagnostic marker for lung cancer, we performed reverse-phase protein array analysis with samples of 195 lung cancer patients and 100 healthy controls. The CANX expression levels were significantly higher in lung cancer patients than in healthy controls (P<0.0001), and the area under the curve of ROC was 0.980, with 96.9% specificity and 99.0% sensitivity. Furthermore, since CANX was also detected in stage I disease, the serum CANX levels should be applicable markers discriminating lung cancer patients from healthy controls and possibly used in the detection of early lung cancer. To our knowledge, the present results provide evidence that CANX may be a novel sero-diagnostic marker for lung cancer.

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“…S100A16, an S100 protein, located in the 1q21 region of human chromosome, was isolated from astrocytoma. S100A16 is widely expressed in specific tissue types such as adipose tissue and brain [ 12 ]. Additionally, S100A16 is strongly associated with the prognoses of several tumors.…”

Section: Introductionmentioning

confidence: 99%

Calbindin S100A16 Promotes Renal Cell Carcinoma Progression and Angiogenesis via the VEGF/VEGFR2 Signaling Pathway

Wang

Tang

et al. 2022

Contrast Media & Molecular Imaging

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Purpose. Recent research has indicated that the calcium-binding protein S100A16 promotes carcinogenesis and tumor growth in several forms of cancer. e objective of this study was to examine the relationship between S100A16 and renal cell cancer. Methods. By using e Cancer Genome Atlas (TCGA) database, the differentially expressed gene S100A16 was identified, and its appearance and link to the prognosis of persons with renal cancer were confirmed. Cox regression was used in multivariate analysis, and a nomogram was developed for internal validation. e correlation between S100A16 and immune cells was analyzed in the TIMER database. Moreover, the potential mechanism of action was investigated utilizing GO and KEGG enrichment analyses. Proliferation, migration, and angiogenesis were investigated in vitro, and the involvement of S100A16 in the undesirable biological events of renal cell carcinoma (RCC) was further explored. Results. S100A16 was the differentially expressed molecule identified through database screening. Malignant tissues showed higher S100A16 expression than noncancerous tissues, and S100A16 expression was mostly localized in the cytoplasm. According to the TCGA and KM-plotter datasets, patients with RCC and low S100A16 expression had superior OS, PFI, and DSS. e C-index of the nomogram was 0.754 (0.726-0.782), and the accuracy of the prediction model was high. e TIMER database shows that the expression of S100A16 is associated with immune infiltration and may play an important role in promoting tumor cell immune escape in the RCC tumor microenvironment. S100A16 may influence the biological processes of RCC via the VEGF/VEGFR2 signaling route and PI3K-Akt signaling pathway and through P53 alteration and cell cycle according to the gene enrichment technique. In vitro cytological experiments demonstrated that S100A16 knockdown can inhibit the proliferation and migration of renal cancer cells and the expression levels of VEGF, VEGFR2, and phosphorylated AKT within renal cancer cells, thereby inhibiting angiogenesis in renal cancer cells and resulting in a poor prognosis of RCC. Conclusion. A decrease in S100A16 expression may dramatically increase the OS, PFI, and DSS of patients with RCC and may thus be used as a biomarker for predicting RCC. It may be associated with the immune infiltration of RCC and play a crucial role in the immune evasion of tumor cells within the RCC microenvironment. Intervention of s100a16 can promote the progression and angiogenesis of renal cell carcinoma through the VEGF/VEGFR2 signal transduction pathway and lead to poor prognosis of renal cell carcinoma. ese findings suggest a potential target for the development of anticancer strategies for renal cancer.

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Acquisition of useful sero-diagnostic autoantibodies using the same patients’sera and tumor tissues

Cited by 8 publications

References 25 publications

S100A16 is a Prognostic Marker for Lung Adenocarcinomas

S100A16 is a Prognostic Marker for Lung Adenocarcinomas

Calnexin is a novel sero-diagnostic marker for lung cancer

Calbindin S100A16 Promotes Renal Cell Carcinoma Progression and Angiogenesis via the VEGF/VEGFR2 Signaling Pathway

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