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ABSTRACT: Triptolide is a diterpenoid triepoxide derived from the traditional Chinese medical herb Tripterygium wilfordii. In the present study, we demonstrated that this phytochemical attenuated colon cancer growth in vitro and in vivo. Using a proteomic approach, we found that 14-3-3 epsilon, a cell cycle- and apoptosis-related protein, was altered in colon cancer cells treated with triptolide. In this regard, triptolide induced cleavage and perinuclear translocation of 14-3-3 epsilon. Taken together, our findings suggest that triptolide may merit investigation as a potential therapeutic agent for colon cancer, and its anticancer action may be associated with alteration of 14-3-3 epsilon.
Cell Biochemistry and Function 02/2012; 30(4):271-8. · 1.77 Impact Factor
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Hui Wang,
Haoran Huang,
Weidong Li,
Xuejun Jin,
Jian Zeng, Yawei Liu,
Ye Gu,
Xuegang Sun,
Ge Wen,
Yanqing Ding,
Liang Zhao
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ABSTRACT: 14-3-3ε regulates diverse biological processes and plays a significant role in the formation of malignant tumors. However, the localization and clinical significance of 14-3-3ε in colorectal cancer (CRC) have not been elucidated.
We investigated 14-3-3ε expression and its prognostic significance in CRC. CRC surgical samples were taken from 137 clinicopathologically characterized CRC cases. 14-3-3ε expression was tested by immunohistochemical assay. Separate Western blot of nuclear and cytosol preparations confirmed nuclear localization of 14-3-3ε protein.
Nuclear expression of 14-3-3ε was observed in 76.9% of normal colorectal tissue and 78.8% of all CRC samples. Statistical analysis showed that there was significant difference of nuclear 14-3-3ε expression in patients categorized according to lymph node metastasis. A trend was identified between decreasing nuclear 14-3-3ε expression in CRC and worsening clinical prognosis. Multivariate analysis showed that loss of nuclear 14-3-3ε expression was an independent prognostic indicator for patient's survival.
The current data provide evidence that 14-3-3ε is not exclusively a cytosolic protein, but is also detectable within the nucleus. Our results suggest that nuclear 14-3-3ε as a suppressor may serve as important biomarker of tumor metastasis. Loss of nuclear 14-3-3ε is closely associated with poor overall survival in CRC patients.
Journal of Surgical Oncology 11/2011; 106(3):224-31. · 2.10 Impact Factor
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ABSTRACT: T lymphoma invasion and metastasis 1 (Tiam1) is a metastasis-related gene of T lymphoma that is also involved in the metastasis of a variety of other cancers, but little research has been done yet to test its usefulness as a prognostic marker in renal cell carcinoma (RCC). In the present study, we investigated Tiam1 expression and its prognostic significance in RCC. RCC surgical tissue samples were taken from 136 patients with RCC who had been followed up for 5 years. The Tiam1 expressions in the 136 clinical samples were tested by immunohistochemical assay to find their association with prognostic significance. We found that 93 of 136 (68.4%) paraffin-embedded archival RCC biopsies showed positive expression of Tiam1. Statistical analysis showed that there was significant difference of Tiam1 overexpression in patients categorized according to lymph node metastasis. Patients with higher Tiam1 expression had shorter overall survival time, whereas patients with lower Tiam1 expression had better survival. Multivariate analysis showed that Tiam1 overexpression was an independent prognostic indicator for patient's survival. Our results suggest that Tiam1 protein is a valuable marker of RCC progression. High Tiam1 expression is associated with poor overall survival in patients with RCC.
Journal of Cancer Research and Clinical Oncology 03/2011; 137(3):393-8. · 2.56 Impact Factor
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ABSTRACT: Oxidative stress is a threat to mammalian cells. To better understand the molecular response and mechanism underlying oxidative stress, we applied two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-fight mass spectrometry analysis to identify differential nuclear protein profiling of mouse fibroblast NIH-3T3 cells exposed to mild-H₂O₂. Thirteen differentially expressed proteins were identified by MS and two of them were further validated by Western blot. The results revealed that exposure to mild-H₂O₂ for 12 h cause up-regulated expression of DJ-1, glutathione S-transferase P 1, DNA ligase I, dynamin 2, nucleophosmin, and down-regulated expression of nucleoside diphosphate kinase A, enolase-α, barrier-to-autointegration factor 1, metastasis associated protein 1, glycosytransferase-like domain containing protein 1, synaptonemal complex protein 1, alpha-centractin, bromodomain, and PHD finger containing 1 (BRPF1). Most of the identified proteins are supported as nuclear proteins localized by previous research. The findings may provide some clues to elucidate cell responses to H₂O₂ and the potential mechanism underlying protection against oxidative stress in fibroblast cells.
Cell Biochemistry and Function 10/2010; 28(7):578-84. · 1.77 Impact Factor
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ABSTRACT: Gut-derived endotoxin and pathogenic bacteria have been proposed to be an important causative factor of morbidity and death during heatstroke. However, the molecular changes underlying heat stress-induced small intestine lesions have not yet been well characterized.
A heatstroke model was established in mice, and the thermal response and pathologic changes in the small intestine were examined during heat stress. Proteins extracted from the small intestine of the heated and control mice were separated by two-dimensional (2D) gel electrophoresis, and different protein spots were further identified by peptide mass fingerprint. Targeted proteins were further verified by Western blot and immunohistochemistry analysis.
Pathologic changes in the small intestine during heat stress were found to be substantial. Using 2 D gel proteomics we identified 14 proteins that were regulated differentially in the small intestine of the mice subjected to heat stress. These 14 identified proteins, seven were down-regulated and the other seven were up-regulated, appeared to be involved in metabolism, chaperone, cell skeleton, defense, signal transduction, DNA repair, and recombination. Using Western blot and immunohistochemical analysis, we further confirmed that down-regulated expression of intestinal fructose 1,6-bisphosphatase (FBP) correlated to the severity of small intestine lesions during heat stress and cooling treatment.
Our results identified 14 differentially expressed proteins, which may contribute to the understanding of molecular mechanisms underlying intestinal injury during heatstroke. Furthermore, intestinal FBP, one of the seven down-regulated proteins, may function as a potential marker for prognosis of gut dysfunction.
Journal of Surgical Research 10/2010; 173(2):332-40. · 2.25 Impact Factor
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Liang Zhao,
Hui Wang,
Chao Liu, Yawei Liu,
Xiaoyan Wang,
Shuang Wang,
Xuegang Sun,
Jianming Li,
Yongjian Deng,
Yong Jiang,
Yanqing Ding
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ABSTRACT: LIM and SH3 protein 1 (LASP-1), initially identified from a cDNA library of metastatic axillary lymph nodes of breast cancer patients, is a specific focal adhesion protein involved in numerous biological and pathological processes. The overexpression of LASP-1 has been described in several types of cancers, but the role of LASP-1 in colorectal cancer (CRC) is unknown. In a previous study, comparative proteomic analysis was performed and LASP-1 was identified as a CRC-associated protein in those patients with CRC.
Using immunohistochemistry, we analysed LASP-1 protein expression in 126 clinicopathologically characterised CRC cases. Using gene transfection and RNA interference, we investigated the effects of LASP-1 overexpression and depletion on tumor cellular behavior in vitro and in vivo. Using 2-D DIGE, we analysed the effect of the presence and absence of LASP-1 gene on protein expression profiles of CRC cells.
Overexpression of LASP-1 was found in metastatic CRC tissues (p=0.002), and its expression level was closely correlated with overall survival of patients with CRC (p=0.002). RNA interference-mediated silencing of the LASP-1 gene in SW620 CRC cells inhibited cell proliferation and migration significantly. However, gene transfection-mediated overexpression of LASP-1 in SW480 CRC cells resulted in aggressive phenotypes of cancer cells and promoted cancer growth and metastasis. Furthermore, both overexpression and silencing of the LASP-1 gene caused a very similar protein expression pattern in different CRC cell lines. The identified LASP-1-modulated proteins, including some key cellular molecules, were involved in various biological processes.
The results show that LASP-1 might be a promising target in the treatment of patients with CRC with growth and metastasis of CRC.
Gut 09/2010; 59(9):1226-35. · 10.11 Impact Factor
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ABSTRACT: Propofol, an anesthetic drug, has been shown to exhibit antioxidant and anti-inflammatory properties in vitro and in vivo. Hypercoagulopathy is a common clinical feature of sepsis, but the effects of propofol on the coagulation system in septic conditions are unclear. Using the gel-based comparative proteomic approach, together with Western blot analysis, ELISA, antithrombin III activity assay, and blood coagulation test, the effect of propofol on serum proteomic profiles in endotoxemic rats was examined. We identified that serum platelet factor-4 (PF4), an endogenous pro-coagulant, was up-regulated in LPS-challenged rats (p<0.001). Endotoxemia also resulted in hypercoagulopathy as evidenced by the shortening of thromboplastin time and thrombin time. Administration of propofol attenuated LPS-stimulated PF4 release and partially reversed the effect of LPS on thromboplastin time (p=0.0012) and thrombin time (p=0.0072). We demonstrated that propofol reduces serum levels of PF4 and partially corrects the hypercoagulopathy associated with endotoxemia in rats.
Biochimica et Biophysica Acta 09/2010; 1804(9):1895-901. · 4.66 Impact Factor
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Jinghua Liu,
Shanchao Zhao,
Jing Tang,
Zhijie Li,
Tianyu Zhong, Yawei Liu,
Dengyu Chen,
Mingzhe Zhao,
Yusheng Li,
Xiaowei Gong,
Peng Deng,
Jiang H Wang,
Yong Jiang
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ABSTRACT: It has been well documented that both endogenous inflammatory mediator advanced glycation end products (AGEs) and exogenous inflammatory inducer lipopolysaccharide play key roles in the initiation and development of inflammatory diseases. However, the combined inflammation-stimulatory effect of AGEs and lipopolysaccharide on endothelial cells, and, furthermore, the underlying signal transduction pathways involved, have not been fully elucidated. We found that in vitro co-stimulation with AGE-human serum albumin (HSA) and lipopolysaccharide exhibits a synergistic effect on proinflammatory cytokine/chemokine interleukin-6, interleukin-8 and monochemoattractant protein-1 production in human umbilical vein endothelial cells. Similar to lipopolysaccharide, AGE-HSA stimulation induced mitogen-activated protein kinase phosphorylation and nuclear factor-kappaB nuclear translocation in human umbilical vein endothelial cells, which was further enhanced by a combination of the two stimulants. Pharmacological inhibitions of each individual signaling pathway, including p38, extracellular signal-regulated kinase 1/2, Jun N-terminal kinase and nuclear factor-kappaB, revealed that activation of all of these four pathways is necessary for the effective induction of interleukin-6, interleukin-8 and monochemoattractant protein-1 by both AGE-HSA and lipopolysaccharide. These results suggest that AGEs and lipopolysaccharide cooperatively induce proinflammatory cytokine/chemokine production by activating mitogen-activated protein kinases and nuclear factor-kappaB in endothelial cells, thus amplifying the inflammatory response and resulting in tissue damage.
FEBS Journal 08/2009; 276(16):4598-606. · 3.79 Impact Factor
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ABSTRACT: The GDP dissociation inhibitors (GDIs) are pivotal regulators of Rho GTPases, which are essential for tumor progression, particularly in the area of metastasis. One member of GDIs was identified as RhoGDI (Rho GDP-dissociation inhibitor alpha, or RhoGDIalpha), but little is known about this protein in tumors. In this study, we used comparative proteomic analysis to show that RhoGDI is markedly up-regulated in metastatic colorectal cancer (CRC). The elevated level of RhoGDI protein in metastatic CRC was confirmed by Western blot at the tissue ( n = 24) and cell ( n = 6) levels. Further, we analyzed RhoGDI protein expression in 126 clinicopathologically characterized CRC cases by immunohistochemistry. Statistical analysis showed that there were significant differences of RhoGDI overexpression in patients categorized according to tumor invasion ( p = 0.018), lymph node metastasis ( p = 0.001) and clinical stage ( p = 0.009). A trend was also identified between high expression of RhoGDI and shorter overall survival ( p = 0.013). In the present work, we also analyzed the effect of RhoGDI on CRC cell line. Gene transfection-mediated overexpression of RhoGDI in HT29 cells, containing a low detectable level of endogenous RhoGDI, resulted in a significant increase in cell proliferation and motility in vitro. These data suggest that RhoGDI may promote CRC progression and metastasis by stimulating tumor cell growth and migration.
Journal of Proteome Research 09/2008; 7(9):3994-4003. · 5.11 Impact Factor
