Current Location:home > Browse

1. chinaXiv:201605.01477 [pdf]

Quantitative Analysis of Integrin alpha 6 in Bladder Cancer Cell Lines

Lu Wei; Yang Gang-Long; Liu Chang-Mei; Guan Feng; Xue Peng
Subjects: Biology >> Biophysics >> Biochemistry & Molecular Biology

The integrins, a family of transmembrane proteins, function in cell-to-cell and cell-to-extracellular matrix (ECM) adhesive interactions, and influence cell signaling of cell growth and differentiation. Expression of integrin 6 in three bladder cancer cell lines, HCV29, KK47 and YST1 were quantitatively analyzed by LC-MS using stable isotope labeling by amino acids in cell culture (SILAC), a simple and powerful proteomic strategy. The results showed that the non-invasive bladder cancer cell line KK47 expressed the highest level of integrin alpha 6. The expression of integrin alpha 6 in invasive bladder cancer cell line YTS1 was also higher than in normal bladder epithelial cell line HCV29. Furthermore, these results were confirmed by Western blotting, qPCR, immunohistochemistry and flow cytometry. Clinical data of mRNA 1TGA6 expression pattern from open-access database ( showed the same result during bladder cancer progression. All these indicated that integrin alpha 6 is associated with the invasion progress of the bladder cancer. The preliminary data in this study may sparkle the fundamental role of integrin 6 in the research of bladder cancer.

submitted time 2016-05-12 Hits644Downloads421 Comment 0

2. chinaXiv:201605.01458 [pdf]

Quantitative Analysis of Differential Proteome Expression in Bladder Cancer vs. Normal Bladder Cells Using SILAC Method

Yang, Ganglong; Lu, Wei; Guo, Jia; Guan, Feng; Xu, Zhipeng; Li, Xiang; Sun, Chengwen; Xue, Peng
Subjects: Biology >> Biophysics

The best way to increase patient survival rate is to identify patients who are likely to progress to muscle-invasive or metastatic disease upfront and treat them more aggressively. The human cell lines HCV29 (normal bladder epithelia), KK47 (low grade nonmuscle invasive bladder cancer, NMIBC), and YTS1 (metastatic bladder cancer) have been widely used in studies of molecular mechanisms and cell signaling during bladder cancer (BC) progression. However, little attention has been paid to global quantitative proteome analysis of these three cell lines. We labeled HCV29, KK47, and YTS1 cells by the SILAC method using three stable isotopes each of arginine and lysine. Labeled proteins were analyzed by 2D ultrahigh-resolution liquid chromatography LTQ Orbitrap mass spectrometry. Among 3721 unique identified and annotated proteins in KK47 and YTS1 cells, 36 were significantly upregulated and 74 were significantly downregulated with >95% confidence. Differential expression of these proteins was confirmed by western blotting, quantitative RT-PCR, and cell staining with specific antibodies. Gene ontology (GO) term and pathway analysis indicated that the differentially regulated proteins were involved in DNA replication and molecular transport, cell growth and proliferation, cellular movement, immune cell trafficking, and cell death and survival. These proteins and the advanced proteome techniques described here will be useful for further elucidation of molecular mechanisms in BC and other types of cancer.

submitted time 2016-05-12 Hits411Downloads258 Comment 0

3. chinaXiv:201605.01413 [pdf]

Lysine Malonylation Is Elevated in Type 2 Diabetic Mouse Models and Enriched in Metabolic Associated Proteins

Du, Yipeng; Zhou, Bo; He, Xiaolong; Wei, Peng; Liu, Pingsheng; Wei, Taotao; Cai, Tanxi; Xue, Peng; Yang, Fuquan; Cai, Tanxi; Xue, Peng; Yang, Fuquan; Li, Tingting; Cai, Tanxi; Zhou, Bo; He, Xiaolong; Wei, Peng
Subjects: Biology >> Biophysics

Protein lysine malonylation, a newly identified protein post-translational modification (PTM), has been proved to be evolutionarily conserved and is present in both eukaryotic and prokaryotic cells. However, its potential roles associated with human diseases remain largely unknown. In the present study, we observed an elevated lysine malonylation in a screening of seven lysine acylations in liver tissues of db/db mice, which is a typical model of type 2 diabetes. We also detected an elevated lysine malonylation in ob/ob mice, which is another model of type 2 diabetes. We then performed affinity enrichment coupled with proteomic analysis on liver tissues of both wild-type (wt) and db/db mice and identified a total of 573 malonylated lysine sites from 268 proteins. There were more malonylated lysine sites and proteins in db/db than in wt mice. Five proteins with elevated malonylation were verified by immunoprecipitation coupled with Western blot analysis. Bioinformatic analysis of the proteomic results revealed the enrichment of malonylated proteins in metabolic pathways, especially those involved in glucose and fatty acid metabolism. In addition, the biological role of lysine malonylation was validated in an enzyme of the glycolysis pathway. Together, our findings support a potential role of protein lysine malonylation in type 2 diabetes with possible implications for its therapy in the future.

submitted time 2016-05-12 Hits885Downloads548 Comment 0

4. chinaXiv:201605.01401 [pdf]

Pro-inflammatory Macrophages suppress PPAR gamma activity in Adipocytes via S-nitrosylation

Yin, Ruiying; Fang, Li; Li, Yingjia; Wang, Nanping; Yin, Ruiying; Fang, Li; Li, Yingjia; Wang, Nanping; Xue, Peng; Li, Yazi; Chen, Chang; Guan, Youfei; Wang, Nanping; Chang, Yongsheng; Chang, Yongsheng
Subjects: Biology >> Biophysics >> Biochemistry & Molecular Biology

Peroxisome proliferator-activated receptor-gamma (PPAR gamma) is a ligand-activated nuclear receptor and plays an essential role in insulin signaling. Macrophage infiltration into adipose tissue is a character of metabolic inflammation and closely related to insulin resistance in type 2 diabetes. The mechanism by which pro-inflammatory macrophages cause insulin resistance remains to be elucidated. Here we showed that coculture with macrophages significantly suppressed the transcriptional activity of PPAR gamma on its target genes in 3T3-L1 preadipocytes and diabetic primary adipocytes, depending on inducible nitric oxide synthase (iNOS). We further showed that PPAR gamma underwent S-nitrosylation in response to nitrosative stress. Mass-spectrometry and site-directed mutagenesis revealed that S-nitrosylation at cysteine 168 was responsible for the impairment of PPAR gamma function. Extended exposure to NO instigated the proteasome-dependent degradation of PPAR gamma. Consistently, in vivo evidence revealed an association of the decreased PPAR gamma protein level with increased macrophage infiltration in visceral adipose tissue (VAT) of obese diabetic db/db mice. Together, our results demonstrated that pro-inflammatory macrophages suppressed PPAR gamma activity in adipocytes via S-nitrosylation, suggesting a novel mechanism linking metabolic inflammation with insulin resistance. (C) 2015 Elsevier Inc. All rights reserved.

submitted time 2016-05-12 Hits592Downloads395 Comment 0

5. chinaXiv:201605.01392 [pdf]

Proteomic Comparison and MRM-Based Comparative Analysis of Metabolites Reveal Metabolic Shift in Human Prostate Cancer Cell Lines

Shu, Qingbo; Cai, Tanxi; Chen, Xiulan; Xue, Peng; Zhu, Nali; Xie, Zhensheng; Wei, Shasha; Niu, Lili; Yang, Fuquan; Shu, Qingbo; Cai, Tanxi; Chen, Xiulan; Xue, Peng; Zhu, Nali; Xie, Zhensheng; Wei, Shasha; Niu, Lili; Yang, Fuquan; Shu, Qingbo; Zhang, Qing
Subjects: Biology >> Biophysics

One of the major challenges in prostate cancer therapy remains the development of effective treatments for castration-resistant prostate cancer (CRPC), as the underlying mechanisms for its progression remain elusive. Previous studies showed that androgen receptor (AR) is crucially involved in regulation of metabolism in prostate cancer (PCa) cells throughout the transition from early stage, androgen-sensitive PCa to androgen-independent CRPC. AR achieves such metabolic rewiring directively either via its transcriptional activity or via interactions with AMP-activated protein kinase (AMPK). However, due to the heterogeneous expression and activity status of AR in PCa cells, it remains a challenge to investigate the links between AR status and metabolic alterations. To this end, we compared the proteomes of three pairs of androgen-sensitive (AS) and androgen-independent (AI) PCa cell lines, namely, PC3-AR(+)/PC3, 22Rv1/Du145, and LNCaP/C42B, using an iTRAQ labeling approach. Our results revealed that most of the differentially expressed proteins between each pair function in metabolism, indicating a metabolic shift between AS and AT cells, as further validated by multiple reaction monitoring (MRM)-based quantification of nucleotides and relative comparison of fatty acids between these cell lines. Furthermore, increased adenylate kinase isoenzyme 1 (AK1) in AS relative to AT cells may result in activation of AMPK, representing a major regulatory factor involved in the observed metabolic shift in PCa cells.

submitted time 2016-05-12 Hits768Downloads451 Comment 0

6. chinaXiv:201605.01292 [pdf]

Comparison of Hydrazide Chemistry and Lectin Affinity Based Enrichment Methods for N-glycoproteomics

Xue Peng; Zhou Yue; Ding Xiang; Wang Jun; Xie Zhen-Sheng; Yang Fu-Quan; Xue Peng; Zhou Yue; Wang Jun; Zhou Yue
Subjects: Biology >> Biophysics >> Biochemistry & Molecular Biology

Glycosylation is one of the most common and important post-translational modifications of proteins. Identification of large-scale N-linked glycoprotein is a very important aspect in glycoproteomics research. The N-glycopeptide enrichment is a key step in high-throughput identification of N-glycosylation site. Lectin enrichment and hydrazide chemistry are the two widely used N-glycopeptides enrichment methods. Each method can only enrich certain types of glycopeptides. It has been reported that the two methods are highly complementary, but few studies compared overlaps of glycosties from the two methods. In this paper, using HepG2 cells, we systematically compared the performance of hydrazide chemistry and lectins enrichment methods. The results showed that although the hydrazide method with glycopeptides enrichment efficiency of 76.7%, far higher than the 54.6% lectin enrichment method, 825 glycoprotein and 1 879 N-glycosylation sites identified with the lectin method was significantly more than 522 glycoprotein and 1 014 glycosylation sites enriched by the hydrazide method. Moreover, the two methods did not show significant complementary, together, only 853 glycoproteins and 1 959 N-glycosylation sites were identified. The overlapping results of identified N-glycosylation sites and N-glycoproteins from the two methods show that lectins enrichment method was better than hydrazide chemistry method.

submitted time 2016-05-11 Hits648Downloads413 Comment 0

  [1 Pages/ 6 Totals]