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Meixiang Li,
Cui Li,
Danjuan Li,
Yuanjie Xie,
Jinfeng Shi,
Guoqing Li,
Yongjun Guan,
Maoyu Li,
Pengfei Zhang,
Fang Peng, Zhiqiang Xiao,
Zhuchu Chen
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ABSTRACT: Recently, the tumor microenvironment is increasingly recognized as playing an important role in cancer proliferation, invasion, and metastasis. To screen stroma-associated proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, laser capture microdissection (LCM) and quantitative proteomic analysis were employed to assess different protein expression of the stroma between NPC and normal nasopharyngeal mucosa (NNM). In this study, periostin was identified to be significantly up-regulated in NPC stroma compared with NNM stroma and the result was further confirmed by Western blotting. Immunohistochemistry showed that over-expression of periostin was frequently observed in the stroma of NPC and matched lymph node metastases (LNM) compared with the stroma of NNM. Statistical analysis showed over-expression of periostin was significantly associated with advanced clinical stage (P < 0.001) and lymph node metastasis (P < 0.001) and decreased overall survival (P < 0.001) in NPC. Cox regression analysis indicated over-expression of periostin was an independent prognostic factor. Furthermore, ectopic expression of periostin was used to examine its effect on invasiveness of NPC cell in vitro and the result showed that periostin was able to promote invasiveness of NPC cell. In conclusion, periostin expression is correlated with tumor stage, lymph node metastasis, and patient survival. Periostin is a potential biomarker for the differentiation and prognosis of NPC, and it might play an important role in the progression of NPC.
Clinical and Experimental Metastasis 06/2012; · 3.52 Impact Factor
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ABSTRACT: To explore whether oncogenes DJ-1 and HSP27 are associated with invasiveness of human pituitary adenoma.
Total proteins were extracted from samples of 20 invasive and 20 non-invasive pituitary adenomas and the expression of DJ-1 and HSP27 was analyzed by Western blot. The correlation of DJ-1and HSP27 with the invasiveness of pituitary adenoma was analyzed.
The strong positive rates of DJ-1 and HSP27 in the 20 invasive pituitary adenoma were 70% (14/20) and 80% (16/20), respectively. The invasive group had significantly higher expression of DJ-1 and HSP27 proteins than the noninvasive group [10% (2/20), 10% (2/20), respectively]. There was a positive correlation between the expression of DJ-1, HSP27 proteins and the invasiveness of pituitary adenoma as judged by the Spearman rank correlation test (P<0.05).
The proliferative activity and abnormal expression of oncogenes DJ-1 and HSP27 may play a significant role in tumorigenesis and progression of pituitary adenoma. There was a significant correlation between the expression of DJ-1 and HSP27 and the invasiveness of pituitary adenoma.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 05/2012; 37(5):481-4.
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ABSTRACT: In this paper, metabolic fingerprints were obtained from 102 nasopharyngeal carcinoma (NPC) patients and 107 healthy adults by gas chromatography-mass spectrometry (GC/MS). Partial least squares-discriminant analysis (PLS-DA) has revealed a pattern recognition discriminating the patients from controls, which sensitivity is 89.72% (96/107) and specificity is 85.29% (87/102). Furthermore, double-blind experiment was carried out and satisfactory results were obtained (total correct rate 87.30%). In addition, metabolites that most strongly influence this separation were obtained. The results indicated that a metabonomic approach is feasible and efficient and deserves further evaluation as a potential novel strategy for the detection of nasopharyngeal carcinoma.
Analytical Letters 05/2011; 44(8):1473-1488. · 1.02 Impact Factor
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ABSTRACT: To provide molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer. Differentially expressed proteins in left sided colon cancer and right sided colon cancer were screened by proteomic technique.
Tissue samples including left sided colon cancer and right sided colon cancer were collected and preserved in the -80 degree refrigerator. In the first part of our experiment, protein was separated by 2-dimensional gel electrophoresis (2-DE) and the images of the gels were acquired by the scanner and then analyzed to find the differentially expression protein-spots in different groups. The peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database. Differentially expressed proteins were assayed by RT-PCR,Western blot, and immunohistochemical method.
Altogether 55 differentially expressed protein spots were screened and 21 spots of them were identified. Compared with the right sided colon cancer, 14 proteins were up-regulated and 7 proteins down-regulated including HSP27 in the left sided colon cancer. HSP27 expressed higher in the right sided colon cancer than in the left sided colon cancer.
There are differentially expressed proteins in left sided colon cancer and right sided colon cancer, especially difference in HSP27 expression at mRNA and protein level, which may be molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 04/2011; 36(4):277-85.
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ABSTRACT: This study aimed to explore the mechanism of multi-drug resistance (MDR) in 5-fluorouracil (5-FU)-induced breast cancer cell MCF-7.
MCF-7 cells were exposed in stepwise escalating concentration of 5-FU to develop the resistant cell line, MCF-7/5-FU. Biological and molecular characteristics of the cells were studied through MTT, flow cytometry, real-time PCR, western-blot, and the global protein profiles between MCF-7/5-FU and parental MCF-7 were compared using proteomic approach. Then some of the differentially expressed proteins were validated by western-blot. In addition, the role of 14-3-3sigma was validated using gene transfection.
Drug resistance of MCF-7/5-FU cells to 5-FU, MX, cDDP, ADM, TAXOL all increased significantly compared with MCF-7 cells and that maybe related to BCRP, but not MDR1 and MRP1. Differentially expressed proteins between MCF-7/5-FU and MCF-7 cells were identified; 12 proteins were up-regulated and 18 proteins were down-regulated in MCF-7/5-FU cells. Expressive levels of some proteins in western-blot validation were consistent with the results in proteomic analysis. Enforced 14-3-3sigma expression can increase the sensitivity of MCF-7/5-FU cells to 5-FU and cDDP.
MDR of MCF-7/5-FU likely associated with differentially expressed proteins and 14-3-3sigma may play a positive role in chemotherapy. These findings may provide theoretical support for the prediction of chemotherapeutic response and reverse of MDR.
Journal of Cancer Research and Clinical Oncology 10/2010; 136(10):1477-88. · 2.56 Impact Factor
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ABSTRACT: To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia.
Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins.
F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.
Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 07/2010; 35(7):641-8.
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Zhefeng Xiao,
Guoqing Li,
Yongheng Chen,
Maoyu Li,
Fang Peng,
Cui Li,
Feng Li,
Yanhui Yu,
Yongmei Ouyang, Zhiqiang Xiao,
Zhuchu Chen
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ABSTRACT: Formalin-fixed, paraffin-embedded (FFPE) tissue specimens represent a potentially valuable resource for protein biomarker investigations. In this study, proteins were extracted by a heat-induced antigen retrieval technique combined with a retrieval solution containing 2% SDS from FFPE tissues of normal nasopharyngeal epithelial tissues (NNET) and three histological types of nasopharyngeal carcinoma (NPC) with diverse differentiation degrees. Then two-dimensional liquid chromatography-tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling was employed to quantitatively identify the differentially expressed proteins among the types of NPC FFPE tissues. Our study resulted in the identification of 730 unique proteins, the distributions of subcellular localizations and molecular functions of which were similar to those of the proteomic database of human NPC and NNET that we had set up based on the frozen tissues. Additionally, the relative expression levels of cathepsin D, keratin8, SFN, and stathmin1 identified and quantified in this report were consistent with the immunohistochemistry results acquired in our previous study. In conclusion, we have developed an effective approach to identifying protein changes in FFPE NPC tissues utilizing iTRAQ technology in conjunction with an economical and easily accessible sample preparation method.
Journal of Histochemistry and Cytochemistry 06/2010; 58(6):517-27. · 2.72 Impact Factor
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ABSTRACT: To determine the significance and expression of S100A9 and NMP238 in cervical carcinoma with different concurrent chemoradiotherapy sensitivities.
Fresh carcinoma tissues were collected from untreated cervical carcinoma patients and preserved at -80 degree. The tissues were classified into 2 groups: a high sensitivity group (HS) and a low sensitivity group (LS) according to their response to concurrent chemoradiotherapy. Protein was separated by 2-dimensional gel electrophoresis (2-DE). Peptide mass fingerprintings (PMF) were acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database. Differential expressed proteins were assayed by Western blot and immunohistochemistry.
Most of the gels were clear and were successfully and reproductively analyzed. Intensity and rate of S100A9 expression were higher in the HS group than in the LS group,and those of NMP238 expression were higher in the LS group than in the HS group.
S100A9 and NMP238 expression is associated with concurrent chemoradiotherapy sensitivity in cervical carcinoma.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 01/2010; 35(1):45-51.
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ABSTRACT: To search for lymph node metastasis-associated proteins in human lung squamous carcinoma (hLSC).
Laser capture microdissection (LCM) was used to purify the target cells from lung primary tumor and matched lymph node metastatic tumor in hLSC. Two-dimensional gel electrophoresis (2-DE) was performed to separate the total proteins of microdissected tumor cells from lung primary tumor and matched lymph node metastatic tumor. PDQuest software was applied to analyze 2-DE images. Differential protein spots between the two types of tissues were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). The expression of Rho-GDIalpha, one of the differential proteins, in the microdissected lung primary tumor cells (LPTC) and matched lymph node metastatic tumor cells (LNMTC) was detected by Western blot.
In the present study, 2-DE patterns of microdissected LPTC and LNMTC were established, and 22 differential proteins in the above two tissues were identified, of which 14 were down-regulated in LNMTC and 8 were up-regulated in LNMTC.
The 22 differential proteins may play some roles in the process of lymph node metastasis in hLSC, and the data provide new clues for metastasis-associated biomarker screen and mechanism of hLSC.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 12/2009; 34(12):1182-8.
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ABSTRACT: To determine the effect of Ad-E1A gene therapy on in vivo radiosensitivity to nasopharyngeal carcinoma.
CNE-2Z cells (2 x 10(5)) were subcutaneously injected into nude mice to develop tumor (1-3 mm) 6 days later. The tumor-bearing mice were then randomly divided into 6 groups (10 mice per group) for PBS treatment or treatment with radiotherapy, Ad-E1A, or Ad-beta-gal alone or radiotherapy in combination with Ad-E1A or Ad-beta-gal. The mice were treated with Ad-E1A or Ad-beta-gal (5 x 10(9) plaque forming units) by intratumoral injection twice weekly for 2 weeks at beginning of week 2. The mice treated with radiotherapy in combination with Ad-E1A or Ad-beta-gal received 2 Gy radiotherapy daily for 5 days following the first week of treatment with Ad-E1A or Ad-beta-gal. Control mice received PBS therapy or radiotherapy only after tumor cells were injected. When the size of tumor exceeded 2 cm, the mice were killed and the tumors underwent immunohistochemical analysis for VEGF and CD34 expression and TUNEL assay for apoptosis.
The growth delay time was longest in the Ad-E1A plus radiotherapy group. Tumors treated with Ad-E1A plus radiotherapy were 4.7-fold smaller than those treated with radiotherapy alone and 5.3-fold smaller than those treated with Ad-E1A alone. The survival rate of tumor-bearing mice treated with Ad-E1A plus radiotherapy was significantly higher than that of other treatment groups. The vessel density and the VEGF expression were significantly lower in tumors treated with Ad-E1A plus radiotherapy than those treated with radiotherapy alone, Ad-E1A alone, Ad-beta-gal alone, or Ad-beta-gal plus radiotherapy (P<0.01). TUNEL staining revealing apoptosis can be detected in the Ad-E1A group, radiotherapy group, Ad-E1A plus radiotherapy group, and more apoptosis can be detected in tumors treated with Ad-E1A plus radiotherapy than those of other treatment groups.
E1A gene therapy can effectively enhance the nasopharyngeal carcinoma sensitivity to the radiotherapy by down-regulating VEGF expression and inducing apoptosis.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 09/2009; 34(8):744-51.
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ABSTRACT: To establish high resolution, reproducible 2-dimensional electrophoresis (2-DE) profiles of invasive and non-invasive pituitary adenoma tissues and to identify differentially expressed proteins between the invasive and non-invasive tissues.
The proteome from invasive and non-invasive pituitary adenomas tissues was dissected and analyzed by: (1) immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis, (2) silver staining, (3) imageMaster 2-D software analysis, (4) peptide mass fingerprint based (PMS) on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and (5) database comparison.
High-resolution 2-D patterns of invasive and non-invasive pituitary adenoma tissues were successfully produced and repeated 3 times for each sample. An average of 1 080+/-24 and 1 035+/-28 spots were detected for invasive and non-invasive pituitary adenoma tissues, respectively. Additionally, 975+/-45 and 918+/-56 spots were found to have an average matching rate of 90.3% and 88.7% for invasive and non-invasive tissues, respectively. The spot positional deviation was (1.563+/-0.259) mm for IEF and (1.088+/-0.206) mm for SDS-PAGE. A total of 99 spots of differential expression were matched between the invasive and non-invasive pituitary adenoma tissues. Thirty differential proteins, some of which were involved in the regulation of cells cycle and signal transduction, were initially characterized by PMS.
The acquisition of well-resolved and reproducible 2-D patterns of invasive and non-invasive pituitary adenoma tissues and the identification of differentially expressed proteins provides a proteome database for invasive pituitary adenomas.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 08/2009; 34(7):569-75.
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ABSTRACT: To establish the protein expression map of nasopharyngeal carcinoma (NPC), and provide a basis for proteomic study of NPC.
Laser capture microdissection (LCM) was used to isolate cancer cells from NPC tissues. Two-dimensional gel electrophoresis(2-DE) was used to separate the total proteins of LCM purified NPC cells. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) was performed to identify the proteins, and bioinformatics was used to construct the 2-DE database of NPC proteome.
A 2-DE reference map of NPC was established. On the 2-DE map, a total of (1 312+/-30) protein spots were detected, and 427 protein spots representing 241 non-redundant proteins were identified. The 2-DE database of NPC proteome was constructed. These data could be accessed at our website (http://www.xyproteomics.org).
A protein expression profile of LCM purified NPC tissues has been established for the first time, which provides useful information and source for proteomic study of NPC.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 07/2009; 34(6):481-6.
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ABSTRACT: This study was designed to use comparative proteomics technology to find the differentially expressed proteins between human lung adenocarcinoma and paired normal tumor-adjacent lung tissues. The total proteins of 20 human lung adenocarcinoma tissues and paired normal tumor-adjacent lung tissues were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE) and Coomassie Blue staining. The differentially expressed proteins were analyzed with image analysis software and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and quadrupole time of flight mass spectrometry (Q-TOF-MS/MS). (1) Well-resolved, highly reproducible 2-DE patterns of human lung adenocarcinoma and paired normal tumor adjacent lung tissues were obtained. (2) PDQUEST 2D image analysis software was used to analysis 20 cases of lung adenocarcinoma and paired normal lung tissues, 1006 +/- 54 spots were matched among tumor tissues and normal lung tissues. Twenty-eight differentially expressed spots were screened. (3) Twenty non-redundant differentially expressed proteins were identified by mass spectrometry, thirteen proteins were up-regulated, and seven proteins were down-regulated, some proteins were involved in the regulation of cell signal transduction or metabolized enzyme of cell. (4) To validate the results screened by proteome research, immunohistochemistry was used to validate several lung adenocarcinoma differentially expressed proteins including 14-3-3 sigma, annexin 1, and manganese superoxide dismutase. The results showed that these three proteins were really differentially expressed between lung adenocarcinoma and paired normal lung tissues. These results will provide scientific foundation and new clue for the research of human lung adenocarcinoma.
Medical Oncology 05/2009; 27(2):346-56. · 2.14 Impact Factor
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ABSTRACT: Nasopharyngeal carcinoma (NPC) is a high-incidence malignancy in Southern China and Southeast Asia. Although mutation of p53 tumor-suppressor gene is a rare event in NPC, NPC has a high frequency of overexpressed/accumulated p53 protein, which was reported to be dysfunction or inactivation in most of NPC. We report here a functional characterization of p53 in an undifferentiated NPC cell line CNE2. To elucidate the biological function of p53, we employed the RNA interference (RNAi) approach to knockdown the endogenously expressed p53 in CNE2 cells. Interestingly, suppression of p53 expression in CNE2 cells was associated with significant down-regulation of p21WAF1/CIP1 expression and decreased HDM2 protein level in both steady state and genotoxic stress induced by ionizing radiation (IR). Consistent with these biochemical data were the accelerated cell cycle progression and the increased proliferation rate, suggesting that p53 retained growth inhibitory activity in CNE2 cells. Indeed, down-regulation of p53 in CNE2 enhanced the ability of CNE2 cells to grow anchorage-independently in vitro and to develop tumors in vivo. Together with the radioresistance acquired by CNE2sip53 cells, our data indicate that in contrast to a previous study, p53 in this NPC cell line remains functional, which may have an important therapeutical implication.
International Journal of Oncology 05/2009; 34(4):1017-27. · 2.40 Impact Factor
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ABSTRACT: To detect the methylation and expression of glioma pathogenesis-related protein 1(GLIPR1) gene in the acute myeloid leukemia (AML) cell lines and bone marrow cells from AML patients, and to determine the relationship between promoter methylation and expression of GLIPR1.
Five leukemia cell lines, 54 bone marrows from the newly diagnosed AML patients, 48 bone marrows from the acute lymphoblastic leukemia (ALL )patients, 40 bone marrows from the chronic myeloid leukemia (CML) patients,35 bone marrows from control patients, and 8 bone marrows from the complete remission AML patients were collected. RT-PCR and methylation-PCR (MSP) were used to detect the mRNA expression and promoter methylation of GLIPR1, respectively, and the relationship between them was analyzed.
The level of GLIPR1 mRNA in the AML cell lines was lower than that in the chronic myeloid leukemia (CML) and ALL cell lines, whereas the methylation level of GLIPR1 in the former was higher than that in the latter. The level of GLIPR1 mRNA in the AML cell lines was significantly increased, but had no obvious changes in the CML and ALL cell lines after 5-aza-2dC treatment. The mRNA level of GLIPR1 in the AML bone marrows (0.38+/-0.20)was obviously lower than that in the ALL bone marrows (0.76+/-0.18), CML bone marrows (0.80+/-0.14), and control bone marrows(0.85+/-0.12). The level of GLIPR1 mRNA in the bone marrows with complete remission AML was obviously higher than that in the AML bone marrows before the treatment (0.78+/-0.13 vs. 0.36+/-0.20); but there was no obvious difference between the ALL bone marrows and the control bone marrows, and the CML bone marrows and the control bone marrows (both P>0.05). The positive rate of GLIPR1 gene methylation in the AML bone marrows (81.5%) was obviously higher than that in the ALL bone marrows(37.5%), CML bone marrows (27.5%) and the control bone marrows(14.3%). The positive rate of GLIPR1 gene in the bone marrows with complete remission AML was obviously lower than that in the bone marrows before the treatment (12.5% vs. 75.0%), but there was no obvious difference between the ALL bone marrows and between the control bone marrows,and the CML bone marrows and the control bone marrows (both P>0.05). There was a negative correlation between the mRNA level and methylation status of GLIPR1 in the AML bone marrows.
GLIPR1 expression is downregulated or even lost by promoter methylation in AML, and the expression and methylation level of GLIPR1 gene may have some significance in evaluating the curative effect of AML patients.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 05/2009; 34(5):388-94.
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Huixin Yao,
Zhiqiang Zhang, Zhiqiang Xiao,
Yongheng Chen,
Cui Li,
Pengfei Zhang,
Meixiang Li,
Yingfu Liu,
Yongjun Guan,
Yanhui Yu,
Zhuchu Chen
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ABSTRACT: A quantitative proteomic approach was used to discover potential protein markers associated with lymph node metastasis (LNM) in human lung squamous carcinoma (LSC). Laser capture microdissection was performed to purify LSC cells with LNM (LNM LSC) and LSC without LNM (non-LNM LSC). The differentially expressed proteins between pooled microdissected non-LNM LSC and LNM LSC cells were identified by two-dimensional difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). 14 proteins were found to be differentially expressed between non-LNM LSC and LNM LSC. Among these proteins, ten proteins were overexpressed in LNM LSC compared with non-LNM LSC, and four proteins were downregulated in LNM LSC. Some of these identified proteins (Annexin A2, HSP27, CK19, and 14-3-3sigma) were further confirmed by Western blotting and immunohistochemical analysis. These results show the value of LCM coupled with 2D-DIGE in identifying potential markers for lymph node metastasis of LSC, and also provide further insights into the prognosis of LSC.
Lung cancer (Amsterdam, Netherlands) 01/2009; 65(1):41-8. · 3.14 Impact Factor
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ABSTRACT: To determine the effect of Ad-E1A gene therapy on the radiosensitivity of nasopharyngeal carcinoma cell by downregulating the expression of VEGF in vitro.
The human nasopharyngeal carcinoma CNE-2Z cell lines were investigated. The recombinant adenovirus vector containing E1A gene was used for this study. After CNE-2Z cells was treated with PBS, Ad-beta-gal and Ad-E1A for 48 h, the three groups were irradiated in different doses at 0, 2.4, 6, 8 and 10 Gy, the cytotoxicity was determined by MTT assay and cell cycle was analysis by flow cytometry. The VEGF expression were evaluated by RT-PCR assay and immunocytochemical analysis.
Significant cell deaths by IR were observed in a dose dependent manner in the three group CNE-2Z cells. After transduction of the E1A gene into CNE-2Z cells, the sensitivity of these cells to radiation was enhanced than the PBS treated group and Ad-beta-gal treated group. Cell growth inhibition in Ad-E1A group by IR was strongly enhanced than Ad-beta-gal treated group and PBS treated group. RT-PCR assay and immunocytochemical analysis showed VEGF expression was downregulated in Ad-E1A treated group.
E1A gene therapy can effectively enhance the nasopharyngeal carcinoma cell sensitivity to the radiotherapy by down-regulating VEGF expression. These findings may pave the way for efficient radiation-gene therapy to NPC in future.
Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 11/2008; 22(20):933-6.
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ABSTRACT: Latent membrane protein 1 (LMP1) of Epstein-Barr virus has been identified to be crucial in inducing cell transformation. However, the mechanism of LMP1-mediated epithelial cell transformation remains unclear. In this study, nasopharyngeal epithelial cells NP69 were infected with retrovirus with gene encoding wild type LMP1 or mutational LMP1 defective in binding to tumor necrosis factor receptor-associated death domain (TRADD). The NP69-LMP1(TRADD) lost some malignant phenotypes compared with the NP69-LMP1(WT). We performed proteomic approach to gain the differential protein expression profile associated with LMP1-mediated epithelial cell transformation. Furthermore, the differential expressional levels of partial identified proteins were confirmed by Western blot and real-time RT-PCR. Some were known to be related to the development of LMP1-induced transformation, and some were new LMP1-associated proteins. These data are valuable for further study of the mechanism of LMP1 in human nasopharyngeal carcinoma and provide some new clues for investigating other LMP1-associated tumors.
Molecular and Cellular Biochemistry 08/2008; 314(1-2):73-83. · 2.06 Impact Factor
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ABSTRACT: Persistent hepatitis C virus (HCV) infection can cause liver cirrhosis and hepatocellular carcinoma. Non-structural protein 3 (NS3), an important part of HCV, has been implicated in the life cycle of the virus and interacts with host cellular proteins. In this study, we investigated the effect of NS3 protein on cell tranformation and related protein alteration in human hepatocyte QSG7701 cells. The results indicated that stable expression of the NS3 protein in QSG7701 cells induced transformed characters with reduced population doubling time, anchorage-independent growth and tumor development. Fifteen differentially-expressed proteins were separated and identified using 2-D electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry. Western blot analysis confirmed that the increase of phospho-p44/42 and phospho-p38 proteins was associated with transformed cells. These results supported the view that HCV NS3 protein plays a transforming role and provided some clues to elucidate the carcinogenesis mechanism of HCV-related hepatocellular carcinoma.
Acta Biochimica et Biophysica Sinica 11/2007; 39(10):751-62. · 1.38 Impact Factor
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ABSTRACT: In order to conduct a comparative proteomics study of human nasopharyngeal carcinoma (NPC) to understand the molecular mechanisms that participate in the formation of NPC, the two-dimensional gel electrophoresis (2-DE) reference map of human NPC tissue proteome was described. To provide a high level of reproducibility between gels and accurately array each protein expressed in NPC tissue proteome, the two-dimensional polyacrylamide gel electrophoresis system, modified colloidal Coomassie Brilliant Blue staining method and ImageMaster 2D Platinum image analysis software were used. The NPC 2-DE maps show that high quality and good reproducibility of the 2-DE gel pattern was attained. An average total of 1,100 protein spots were separated by 2-DE, visualized by a modified colloidal Coomassie Brilliant Blue staining method. A synthesized 2-DE reference gel was acquired after detailed analysis of the NPC 2-DE gel maps, and 216 medium to high abundant spots were identified as landmark spots of NPC 2-DE gel, which expressed on >75% of gels. To provide an unambiguous identification of the landmark spots in gels, MALDI-TOF, ESI-Q-TOF mass spectrometry and database search were used to identify the proteins expressed in NPC tissue proteome. Between the 216 landmark spots, all proteins were identified with MALDI-TOF at first, 41 of which were identified with both MALDI-TOF and ESI-Q-TOF. All identified proteins were classified in terms of their subcellular localization and physiological function with information from SWISS-PROT and NCBI websites. According to our knowledge this is the first 2-DE reference map of human NPC. This reference map will serve as a basis for further studies of human NPC and the reference map data will be used to expand the proteome database of human NPC, which can be accessed in our website (http://www.xyproteomics.org/).
International Journal of Oncology 05/2007; 30(5):1077-88. · 2.40 Impact Factor
