期刊：
BioMed Research International,2012年2012(3):510418-510429 ISSN：2314-6133
通讯作者：
Li Cui
作者机构：
[Li Cui; Chen Zhu-Chu; Xiao Zhi-Qiang; Xu Yan; Peng Fang; Duan Chao-Jun; Tang Can-E] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Prote, Changsha 410008, Hunan, Peoples R China.;[Xu Yan] Judicial Police Gen Hosp, Changsha 410004, Hunan, Peoples R China.;[Cao Lan-Qin] Cent S Univ, Xiangya Hosp, Dept Gynecol & Obstet, Changsha 410008, Hunan, Peoples R China.;[Jin Long-Yu] Cent S Univ, Xiangya Hosp 3, Dept Cardiothorac Surg, Changsha 410013, Hunan, Peoples R China.;[Zeng Gu-Qing] Univ S China, Sch Med, Dept Gen Intro Surg, Hengyang 421001, Peoples R China.
通讯机构：
[Li Cui] C;Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Prote, Changsha 410008, Hunan, Peoples R China.
摘要：
Objective. To investigate the differential protein profile of human lung squamous carcinoma (HLSC) and normal bronchial epithelium (NBE) and provide preliminary results for further study to explore the carcinogenic mechanism of HLSC. Methods. Laser capture microdissection (LCM) was used to purify the target cells from 10 pairs of HLSC tissues and their matched NHBE, respectively. A stable-isotope labeled strategy using iTRAQ, followed by 2D-LC/Q-STAR mass spectrometry, was performed to separate and identify the differential expression proteins. Results. A total of 96 differential expression proteins in the LCM-purified HLSC and NBE were identified. Compared with NBE, 49 proteins were upregulated and 47 proteins were downregulated in HLSC. Furthermore, the expression levels of the differential proteins including HSPB1, CKB, SCCA1, S100A8, as well as S100A9 were confirmed by western blot and tissue microarray and were consistent with the results of quantitative proteomics. Conclusion. The different expression proteins in HLSC will provide scientific foundation for further study to explore the carcinogenic mechanism of HLSC.
摘要：
To identify a novel lung adenocarcinoma (AdC) biomarker, iTRAQ-tagging combined with 2D LC-MS/MS analysis was used to identify differentially expressed plasma membrane (PM) proteins in primary lung AdCs and paraneoplastic normal lung tissues (PNLTs). As a result, 36 differentially expressed membrane proteins were identified. Two differential PM proteins flotillin-1 and caveolin-1 were selectively validated by Western blotting. As there has been no report on the association of flotillin-1 with lung AdC, immunohistochemistry was further performed to detect the expression of flotillin-1 in the archival tissue specimens including 42 cases of PNLTs, 62 cases of primary, lung AdCs with lymph node metastasis (LNM AdCs), and 46 cases of primary lung AdCs without lymph node metastasis (non-LNM AdCs), and the correlation of flotillin-1 expression levels in lung AdCs with clinicopathological features and clinical outcomes were evaluated. The results showed that up-regulation of flotillin-1 expression in lung AdCs was significantly correlated with advanced clinical stage, lymph node metastasis, increased postoperative relapse and decreased overall survival. Cox regression analysis revealed that the expressional level of flotillin-1 was an independent prognostic factor. The data suggest that flotillin-1 is a potential novel biomarker for lymph node metastasis and prognosis of lung AdC, and flotillin-1 up-regulation might play an important role in the pathogenesis of lung AdC. (C) 2012 Elsevier B.V. All rights reserved.
摘要：
Mutations in the PKHD1 gene result in autosomal recessive polycystic kidney disease (ARPKD) in humans. To determine the molecular mechanism of the cystogenesis in ARPKD, we recently generated a mouse model for ARPKD that carries a targeted mutation in the mouse orthologue of human PKHD1. The homozygous mutant mice display hepatorenal cysts whose phenotypes are similar to those of human ARPKD patients. By littermates of this mouse, we developed two immortalized renal collecting duct cell lines with Pkhd1 and two without. Under nonpermissive culture conditions, the Pkhd1(-/-) renal cells displayed aberrant cell cell contacts and tubulomorphogenesis. The Pkhd1(-/-) cells also showed significantly reduced cell proliferation and elevated apoptosis. To validate this finding in vivo, we examined proliferation and apoptosis in the kidneys of Pkhd1(-/-) mice and their wildtype littermates. Using proliferation (PCNA and Histone-3) and apoptosis (TUNEL and caspase-3) markers, similar results were obtained in the Pkhd1(-/-) kidney tissues as in the cells. To identify the molecular basis of these findings, we analyzed the effect of Pkhd1 loss on multiple putative signaling regulators. We demonstrated that the loss of Pkhd1 disrupts multiple major phosphorylations of focal adhesion kinase (FAK), and these disruptions either inhibit the Ras/C-Raf pathways to suppress MEK/ERK activity and ultimately reduce cell proliferation, or suppress PDK1/AKT to upregulate Bax/caspase-9/caspase-3 and promote apoptosis. Our findings indicate that apoptosis may be a major player in the cyst formation in ARPKD, which may lead to new therapeutic strategies for human ARPKD. (C) 2010 Elsevier Inc. All rights reserved.
摘要：
Radiotherapy is the primary treatment for nasopharyngeal cancer (NPC), and p53 is closely associated with the radiosensitivity of cancer, but the molecular mechanisms of p53-mediated radioresponse in NPC remains unclear. We previously established NPC CNE2sip53 cell line with p53 knockdown and paired control cell line CNE2/pSUPER, which provides a cell model system to investigate mechanisms of p53-mediated radioresponse in NPC. In this study, we first compared the radiosensitivity of CNE2sip53 and CNE2/pSUPER by a clonogenic survival assay, cell growth assay, and Hoechst 33258 staining and flow cytometry analysis of apoptotic cells. The results showed that the radiosensitivity of CNE2sip53 was significantly lower than that of CNE2/pSUPER, indicating that p53 plays a role in mediating NPC radiosensitivity. To search for the proteins associated with the p53-mediated radioresponse in NPC, a proteomic approach was performed to identify the radioresponsive proteins in CNE2sip53 and CNE2p/SUPER, respectively, and then the difference of radioresponsive proteins in CNE2sip53 and CNE2p/SUPER was compared. As a result, 14 differential radioresponsive proteins were identified in the two cell lines, 4 proteins of which were conformed by Western blot. Among them, 9 and 5 proteins were identified solely from CNE2p/SUPER and CNE2sipS3, respectively. Furthermore, protein protein interaction analysis showed that 7 differential radioresponsive proteins identified only in CNE2p/SUPER were related to p53 protein. Our results suggest that the differential radioresponsive proteins unique to CNE2p/SUPER may be involved in p53-mediated radioresponse in NPC, which will be helpful for elucidating the mechanisms of p53-mediated NPC cellular response to radiotherapy. (C) 2011 Elsevier B.V. All rights reserved.
期刊：
Medical Oncology,2010年27(1):134-144 ISSN：1357-0560
通讯作者：
Chen, Zhu-chu
作者机构：
[Yao, Hui-xin; Li, Dan-Juan; Li, Cui; Xiao, Zhi-qiang; Li, Mao-yu; Duan, Chao-jun; Peng, Fang; Zhang, Peng-fei; Li, Mei-xiang; Chen, Zhu-chu] Cent S Univ, Key Lab Canc Prote, Chinese Minist Hlth, Xiangya Hosp, Changsha 410008, Hunan, Peoples R China.;[Yao, Hui-xin; Li, Feng; Li, Mei-xiang; Chen, Zhu-chu] Cent S Univ, Canc Res Inst, Xiangya Sch Med, Changsha 410078, Hunan, Peoples R China.;[Li, Mei-xiang] Univ S China, Dept Histol & Embryol, Hengyang 421001, Hunan, Peoples R China.;[Chen, Yong-heng] Univ So Calif, Los Angeles, CA USA.;[Chen, Zhu-chu] Cent S Univ, Key Lab Canc Prote, Chinese Minist Hlth, Xiangya Hosp, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
通讯机构：
[Chen, Zhu-chu] C;Cent S Univ, Key Lab Canc Prote, Chinese Minist Hlth, Xiangya Hosp, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
关键词：
Nasopharyngeal carcinoma;Stroma;Laser capture microdissection;Two-dimensional gel electrophoresis;Mass spectrometry
摘要：
The stroma surrounding cancer cell population is increasingly recognized as playing an important role in cancer proliferation, invasion, and metastasis. To identify the stromal proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, differences in protein expression of the stroma from NPC and normal nasopharyngeal epithelium tissues (NNET) were assessed using a comparative proteomic approach combined with laser capture microdissection (LCM). LCM was performed to purify stromal cells from NPC and NNET, respectively. Proteins between the pooled microdissected tumor and normal stroma were separated by two-dimensional electrophoresis (2-DE) and differential proteins were identified by mass spectrometry (MS). Sixty differential proteins between normal stroma (NS) and tumor stroma (TS) were identified, and the expression of CapG protein was further confirmed by western blotting and immunohistochemical analysis. Our results will be helpful to study the role of stroma in the NPC carcinogenesis and may provide helpful clues for pathogenesis, early diagnosis, and progression of NPC.
摘要：
14-3-3 sigma is a potential tumor suppressor, and loss of 14-3-3 sigma expression plays an important role in carcinogenesis and metastasis. To explore the possible mechanism of 14-3-3 sigma in nasopharyngeal carcinoma (NPC) invasion and metastasis, targeted proteomic analysis was performed on 14-3-3 sigma-associated proteins from NPC cells. As the results, 112 proteins associated with 14-3-3 sigma were identified, and four 14-3-3 sigma-interacted proteins: keratin 8, epidermal growth factor receptor (EGFR), small GTP-binding protein RAB7, and p53 were confirmed by coimmunoprecipitation and Western blot analysis. The 14-3-3 sigma-associated proteins could be grouped into eight clusters based on their molecule functions. Protein-protein interaction (PPI) analysis indicated that 14-3-3 sigma/EGFR/keratin 8 interactions may be involved in the invasion and metastasis of NPC. 14-3-3 sigma/EGFR/keratin 8 could form complexes in NPC cells. 14-3-3 sigma downregulation in NPC may lead to the overexpression of EGFR and keratin 8, which increases the invasion ability of NPC cells possibly by activating the downstream signal molecules and reorganizing cytoskeleton. The data suggest that the biological functions of 14-3-3 sigma in NPC are diversified, and 14-3-3 sigma could inhibit the in vitro invasive ability of NPC cells possibly through 14-3-3 sigma/EGFR/keratin 8 interaction. Crown Copyright (C) 2009 Published by Elsevier Ltd. All rights reserved.
作者：
Liu, Y. F.;Xiao, Z. Q.;Li, M. X.;Li, M. Y.;Zhang, P. F.;...
期刊：
JOURNAL OF PATHOLOGY,2009年217(1):54-64 ISSN：0022-3417
通讯作者：
Chen, Z-C
作者机构：
[Li, C.; Zhang, P. F.; Yi, H.; Xiao, Z. Q.; Li, F.; Liu, Y. F.; Yao, H. X.; Chen, Z-C; Li, M. X.; Li, M. Y.] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, Changsha 410008, Hunan, Peoples R China.;[Li, F.; Liu, Y. F.; Yao, H. X.; Chen, Z-C; Li, M. X.] Cent S Univ, Xiangya Sch Med, Canc Res Inst, Changsha 410078, Hunan, Peoples R China.;[Li, M. X.] Univ S China, Dept Histol & Embryol, Hengyang, Peoples R China.;[Chen, Y. H.] Univ So Calif, Los Angeles, CA USA.;[Chen, Z-C] Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
通讯机构：
[Chen, Z-C] C;Cent S Univ, Xiangya Hosp, Chinese Minist Hlth, Key Lab Canc Proteom, 87 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
摘要：
Clinical observations have shown a link for the high comorbid rate between smoking and psychiatric disorders, including anxiety disorders. However, little is known about the neural mechanism underlying the progression from nicotine dependence to an anxiety disorder. A deficit in fear extinction in general is considered to contribute to anxiety disorders. The aim of the present study is to investigate the effects of chronic nicotine on fear extinction in rats. Rats were administrated s.c. nicotine twice per day for 14 days. Two weeks after the last injection rats received a cued or contextual fear conditioning session. Twenty-four hours and 48 h after conditioning, rats received an extinction training session and an extinction test session, respectively. Percent freezing was assessed during all phases of training. In the cued task, prior chronic nicotine did not affect the acquisition of fear response or the within-session fear extinction, but impaired the between-session fear extinction. In the contextual task, the same nicotine treatment schedule did not affect the acquisition of fear response or the within- and between-session fear extinction, but enhanced the retention of fear conditioning. This prior chronic nicotine-induced deficit in cued fear extinction and/or enhanced fear to context may be one of the critical components that contribute to the progression from nicotine dependence to an anxiety disorder.
