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@#Objective To investigate the effects of a disintegrin and metalloproteinase 17(ADAM17) deletion on the production of reactive oxygen species(ROS) and mitochondrial function in nasopharyngeal carcinoma(NPC) cells.Methods Three groups of ADAM1 7 interfering plasmid ADAM17 shRNA and empty plasmid ADAM17-shRNA-NC were transfected into NPC cell line(CNE1) and detected for the interference efficiency by RT-PCR and Western blot to select shRNA with the best interference effect for the follow-up experiments.The cell proliferation was detected by CCK-8 assay,while the cell growth by clone formation test,the apoptosis and changes in mitochondrial membrane potential(MMP) by flow cytometry,the level of mitochondrial oxidative damage product ROS by fluorescence microscope,the contents of oxidative stress markers MDA and SOD by malondialdehyde(MDA) kit and superoxide dismutase(SOD) kit and the expression of mitochondrial damage markers Bax/Bcl-2,cleaved-caspase 9/caspase 9,cleaved-caspase 3/caspase 3 and c-Myc by Western blot.Results ADAM17-shRNA2 group showed the best interference effect.Compared with shRNA-NC group,the proliferation rate of cell in ADAM17-shRNA 2 group decreased significantly(t=8.964,P=0.036);the number of colonies were significantly reduced(t=10.351,P=0.014);the number of apoptosis increased significantly(t=11.25,P=0.008);the fluorescence intensity representing ROS level in cells increased obviously;the mitochondrial membrane potential decreased significantly(t=9.233,P=0.013);the SOD content decreased(t=7.233,P=0.034) and MDA content increased(t=7.415,P=0.038) significantly;the levels of Bax/Bcl-2,cleaved-caspase 9/caspase 9 and cleaved-caspase 3/caspase 3 significantly increased(t=8.985,9.021 and 7.789,P=0.023,0.011 and 0.031,respectively),while the expression of c-Myc proteins significantly decreased(t=10.352,P=0.004).Conclusion Interfering with ADAM1 7 induced SOD decrease and MDA increase by promoting oxidation,thereby alleviating oxidative damage of cell membrane,which also promoted the expression level of ROS in mitochondrion,reduced MMP,inhibited cell proliferation in vitro,and promoted apoptosis.
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The erythropoietin-producing hepatocellular receptor (Eph) and its ligand ephrin are the largest of the receptor tyrosine kinases (RTKs) family in humans. Since ephrin ligands and Eph receptors are membrane-bound proteins, binding and activation of Eph/ephrin intracellular signaling pathways can only occur via direct cell-cell interaction. Eph-ephrin complexes emanate bidirectional signals that affect cells expressing Eph and ephrin, respectively. Its repulsive signaling effects include retraction, which plays an important role in many physiological and pathological processes. EphA2 has been found to have a strong association with tumors and is most widely studied. EphA2 signal transduction in tumor cells may promote or inhibit tumor, depending on the tumor microenvironment. EphA2 "canonical" signaling involves ligand binding and kinase activity; thus EphA2 "noncanonical" signaling is ligand independent and lacks kinase activity. This review summarizes the pathogenesis of EphA2 in nasopharyngeal carcinoma (NPC), including ligand independent signal and EBV infection receptor, furthermore evaluates the prospect of its potential utilization as a target for cancer therapeutics. This may provide a new method for the prevention and treatment of NPC.
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@#[Abstract] Objective: To explore the role of adhesion molecule with Ig like domain 2 (AMIGO2) in the proliferation of nasopharyn‐ geal carcinoma (NPC) cells and its mechanisms. Methods: A total of 10 NPC tissue samples and 10 normal nasopharyngeal epithelial tissue samples collected at Fujian Cancer Hospital during September 2017 and November 2017 were used for this study; in addition, NPC cell lines (CNE-1, CNE-2, SUNE-1, 6-10B, C666-1) and human immobilized nasopharyngeal epithelial cell line NP69 were also collected. The relative expression of AMIGO2 mRNAin above mentioned tissues and cell lines was detected by qPCR. Lentivirus vectors were constructed to interfere AMIGO2 mRNA expression, and qPCR was used to verify its interference efficiency. CCK-8 method, Clonal formation and Flow cytometry were performed to evaluate the effect of AMIGO2 interference on proliferation, clone formation and apoptosis of NPC cells. The influence of AMIGO2 interference on PI3K/AKT/mTOR signaling pathway and proliferation related molecular markers in NPC cells was assessed by Western blotting. Results: The results of qPCR showed that AMIGO2 was highly expressed in NPC tissues, CNE-2, and SUNE-1 cells (all P<0.01). The interference efficiency of AMIGO2 in CNE-2 and SUNE-1 cells could reach over 50%. The interfering of AMIGO2 expression significantly inhibited the proliferation and clone formation of CNE-2 and SUNE-1 cells (all P<0.01), promoted cell apoptosis (all P<0.01), reduced the phosphorylated protein expression levels of PI3K, AKT and mTOR in SUNE-1 cells (all P<0.01), as well as down-regulated the protein expressions of survivin and PCNA (all P<0.01). Conclusion: AMIGO2 may promote the proliferation as well as inhibit apoptosis of NPC cells by activating the PI3K/AKT/mTOR signaling pathway, suggesting that AMIGO2 may be a potential target for NPC therapy.
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Objective: To investigate the efficacy and mechanism of EGDT against NPC cell lines. Methods: MTT assay was used to assess cell proliferation inhibition of EGDT. The apoptotic induction and cell cycle arrest were detected by flow cytometry. Western blot was adopted to detect the protein levels. Quantitative Real-time PCR was used to determine the mRNA expressions. The NPC xenografts were established to evaluate the tumor growth inhibition of EGDT. Immunohistochemistry was applied to analyze the EGFR expression in the tumor tissues. Results: EGDT showed proliferation inhibition on the NPC cell, induced G0/G1 phase arrest and cell apoptosis in vitro. EGDT decreased the protein and mRNA levels of EGFR and its downstream RAF/MEK/ERK and PI3K/AKT pathways in time- and dose-dependent manner. Furthermore, EGDT also showed a sound antitumor activity in NPC xenograft in vivo. Conclusion: The treatment of EGDT displays EGFR and its mediated downstream signaling pathway blockade through decreasing the protein and mRNA levels, suggesting a promising strategy in treating human NPC.
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Objective To investigate the present status of early diagnosis and misdiagnosis of nasopharyngeal carcinoma (NPC). Methods A total of 50 patients diagnosed as NPC were recruited at the department of nasopharyngeal carcinoma in our hospital. The time from that patients felt discomfort to be diagnosed of NPC was determined bydifferent symptoms and signs. Furthermore , the status of misdiagnosis were also investigated. The spearmans rank correlation was used to analyze the correlation between the cancer stage and the time of confirmeddiagnosis , and the relationship between the rate of misdiagnosed and the rank of the hospital they visited. The χ2 test was then used to analyze the cancer stage with the time when they were diagnosed. Results Results indicated that the time when patients were diagnosed significantly correlated with pTNM stage (P < 0.05). Patients diagnosed in one month were most at stage Ⅱ, diagnosed in six months were stage III, and diagnosed after twelve months were stage Ⅳ(P < 0.05). Furthermore, we found that the rate of misdiagnosis of NPC was 12%. Moreover,the misdiagnosed rate was associated with the rank of the hospital patients visited. Discussion In conclusion , the present status of the early diagnosis of NPC is not optimistic. Most of the patients with NPC were misdiagnosed in basic medical institutions , especially in town or village health center. Thus, it is important to popularize the health knowledge about the secondary prevention of NPC and train the doctor in basic medical institutions.
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ABSTRACT:Objective To explore the effects of virus interleukin‐10 (vIL‐10 ) on different expressions of MHC‐I antigen processing “the operon” .Methods We collected nasopharyngeal carcinoma cells (CNE‐1 and CNE‐2) treated by vIL‐10 at different time points ,and detected the changes of MHC‐I antigen processing “the operons” (TAP‐1 ,TAP‐2 ,LMP‐2 ,LMP‐7 and HLA‐I) by RT‐PCR and Western blot .Results ① mRNA level :There was no difference in the expression of TAP‐1 in CNE‐1 and CNE‐2 cells at various time points .The expressions of TAP‐2 and LMP‐2 in CNE‐1 and CNE‐2 did not change at 1 ,4 ,6 ,12 h ,but downregulated and even disappeared at 24 h .The expression of LMP‐7 in CNE‐1 decreased 4 h after vIL‐10 was added ,and that in CNE‐2 decreased at 6 h .The expression of HLA‐I in CNE‐1 and CNE‐2 showed significant decrease at 24 h .② Protein expression :The expression of TAP‐1 in CNE‐1 and CNE‐2 showed significant decrease at 24 h .The expression of TAP‐2 in CNE‐1 and CNE‐2 was gradually downregulated at different time points .The expressions of LMP‐2 and LMP‐7 in CNE‐2 were gradually downregulated at different periods ,while that in CNE‐1 was only decreased at 12 h .The expression of HLA‐I in CNE‐1 and CNE‐2 was gradually downregulated ,but there was no significant difference at each period in CNE‐1 ,while the expression of HLA‐I in CNE‐2 at 24 h was significantly downregulated .Conclusion vIL‐10 can inhibit the expression of MHC‐I antigen processing “the operon” in NPC in the time‐dependent manner .
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Objective:To explore the effect of family history of cancer on clinical features and prognostic factors in nasopharyngeal car-cinoma (NPC) patients. Methods:The clinical data of 89 NPC patients with a family history of cancer and 388 NPC patients without a family history of cancer were retrospectively reviewed. Univariate and multivariate survival analyses were performed to identify possi-ble prognostic factors. Results:The clinical characteristics of NPC patients with and without family history of cancer were compared. The gender, age, TNM stage, pathological type, and hemoglobin radiotherapy concentration before treatment did not significantly dif-fer between the two groups (P>0.05). NPC patients with a family history of cancer had better 3-year overall survival than those with-out family history of cancer (91.6%vs. 85.5%), but no statistically significant difference was observed (P=0.211). Both univariate and multivariate analyses showed that T, N, and TNM stages were the important prognosis factors affecting 3-year overall survival (OS), progression free survival (PFS), and distant metastasis-free survival (DMFS) of NPC (P0.05). Conclusion:NPC patients with family history of cancer had better 3-year OS than those without family history of cancer, but no statistically significant observation was found. Large T stage or high lymph node stage contributed to poor survival of NPC. Family history of cancer had no significant in-fluence on the survival of NPC patients.
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Objective:To retrospectively investigate the regulation of cervical and posterior to level V (PLV) lymph node metastasis on clinical target delineation in radiotherapy for nasopharyngeal carcinoma (NPC). Methods:A total of 428 NPC cases from February 2013 to April 2016 were subjected to enhanced CT scan from the base of the skull to the clavicle for pathological diagnosis. A deputy chief physician and an attending physician assessed the nodal distribution in each level in accordance with the RTOG guidelines proposed in 2013. The central point of the metastatic lymph nodes of PLV in the patients were recreated proportionally on the CT images of a stan-dard patient with N0 NPC in reference to the normal anatomy of the PLV area. SPSS 19.0 was used to analyze the correlation between PLV and the other levels. Moreover, the nodal location and characteristics of PLV were analyzed. Results:Among the 428 patients, 381 (89.0%) showed nodal involvement. The top four metastatic probabilities were presented as follows:Ⅱb (75.2%),Ⅶa (60.3%),Ⅱa (59.6%), andⅢ(42.0%). Up to 21 (4.9%) patients exhibited nodal involvement of PLV with 32 nodes. The mean vertical distance of all central points of PLV from the anterior border of the trapezius was 16 mm. Correlation analysis indicated the nodal involvement of PLV with the ipsilateral level Va (P=0.001). Conclusion:NPC showed a high probability of nodal metastasis. Nodes were mostly metasta-sized from the upper to the lower level, as well as from the proximal to the distal area. The leap metastasis rate was very low. The nod-al involvement of PLV correlated with the ipsilateral metastasis of level Va. Thus, the ipsilateral delineation of the posterior border of level V should be contoured to 25 mm far from the anterior surface of the trapezius during the nodal involvement of level Va.
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Objective To explore the immune function of the whole body and the tumor site of human nasopharyngeal carcinoma (NPC) patients as well as its correlation with CCR4. Methods The ratios of CD4~+CD25~+T cells and CCR4+cells to lymphocytes were measured by flow cytometry in tissues (25 cases) and peripheral blood (35 cases) from nasopharyngeal carcinoma patients, and then statistical analysis was made. Results The ratios of CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC were higher than those in the control group (P<0.05). In NPC the ratios of the two cells in tissue were higher than in peripheral blood respectively (P<0.05), but there was no such difference between tissue and peripheral blood in the control group (P>0.05). The ratio of CD4~+CD25~+T cells in tissue at stage Ⅲ+Ⅳ was higher than that at stage Ⅰ+Ⅱ of NPC (P<0.05), but there was no such difference between the two stages in peripheral blood in NPC. There was a positive correlation between CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC (P<0.05). Conclusion NPC patients have different immunosuppression, and there is even more severe immunosuppression in the tumor site. The ratio of CD4~+CD25~+T cells is correlated with the stage of NPC. Therefore, as NPC progresses to later stages, the percentage of CD4~+CD25~+T cells is increased, which is correlated with poor prognosis. CCR4 plays an important role in reactant of CD4~+CD25~+T cells to tumor sites, and is resistant to immunosurveillance.
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Background and purpose: There have been no reports on the clinical and biological significance of the co-expression of inducible nitric oxide synthase (iNOS) and β-catenin (β-cat) in has.pharyngeal carcinoma(NPC). This study was aimed to investigate the expression of iNOS and β-cat in NPC, to analyze their interrelation, and to explore their roles in the carcinogenesis, development, invasion, and metastasis of NPC. Methods: The expression of iNOS and β-cat was examined in 50 poorly differentiated NPC and 15 normal nasopharyngeal epithelium tissue by immunohistochemical staining (SP method). None of the patients had received radiotherapy and chemotherapy. The results of immunostaining were observed by two pathologists independently, using double blind scoring method. iNOS and β-cat expression were categorized by the extent and intensity of staining using a semiquantitative method. Results: No iNOS expression was observed in 15 normal nasopharyngeal epithelium, but β-cat expression was located in cytomembrane in normal samples. The positive rate of iNOS protein expression was 74.0% (37/50) in NPC tissues. The expression of iNOS was statistically different among the NPC group and normal nasopharyngeal epithelium group, T_(1-2) group and T_(3-4) group, metastatic lymph nodes group and no metastatic lymph nodes group. There was a great quantity of β-cat expression in cytoplasm, little or no β-cat expression in cytomembrane and nucleus in NPC tumorous tissues. Statistical analysis indicated that strong positive expression of β-cat in cytoplasm was significantly higher in NPC than those of normal nasopharyngeal epithelium. The positive expression of β-cat was significantly correlated with clinical stage, invasion and lymph gland metastasis. Neither iNOS nor β-cat was significantly different between different ages and genders. There was a positive relationship between iNOS and β-cat expression in NPC tissues (r=0.394, P=0.005). Conclusion: The positive expressions of iNOS and β-cat in cytoplasm were significantly correlated with clinical stage, invasion and metastasis, and were not significantly different in terms of ages and genders of the patients. There was a positive relationship between the expression of iNOS and β-cat in NPC tissues. Co-overexpression of iNOS and β-cat may play an important role in carcinogenesis, development, invasion, and metastasis of NPC.
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Background and purpose:Concurrent chemo-radiotherapy(CCRT) was considered the best treatment plan for advanced nasopharyngeal carcinoma(NPC),but there was no uniform conclusion as to which category of patients and which chemotherapy associated radiotherapy would have the best therapeutic effect. As the standard treatment plan for advanced NPC, DDP concurrent chemo-radiotherapy was recommended by some scholars. DDP can raise the expression of inducible nitric oxide synthase(iNOS) protein and synthesize nitric oxide (NO) with anti-tumor effects, so we considered whether the therapeutic effect could be predicted and the corresponding treatment plan could be selectived to detect the iNOS expression in the pretherapy NPC tissues.The purpose of this study was to investigate the relation between the expression of iNOS protein and the nasopharyngeal tumor with complete response or with residue after DDP concurrent chemo-radiotherapy, so that the most appropriate plan of treatment can be adopted and the complete response rate of nasopharyngeal tumor can be raised. Methods:All patients were poorly differentiated NPC.The expression of iNOS protein was examined in 30 patients of nasopharyngeal tumor with complete response and 30 patients with residual tumor after DDP concurrent chemo-radiotherapy by immunohistochemical staining (SP method).None of the patients had received radiotherapy and chemotherapy.Results:Immunohistochemical examination revealed that iNOS expression in the NPC tissues was located in both the nucleus and cytoplasm of the tumorous tissues. The intensity of iNOS expression was stronger in the nucleus than in the cytoplasm of the tumorous tissues.The positive rates of iNOS protein expressions were 71.67%(43/60) in NPC tissues. It was 86.67% and 53.33% in 30 tumors with complete response and with residual tumor, respectively. The difference was statistically significant.The rate of iNOS strong postive expressions in the group of residual tumors was higher than that of the group with complete response. It was statistically different,but weak and moderate postive expressions did not have statistical difference.Conclusions:According to the difference of iNOS expression, it is a valuable method to select the most appropriate plan of treatment and the complete response rate of nasopharyngeal tumor can be raised.
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In order to screen EGFR-regulated secreted proteins in human nasopharyngeal carcinoma(NPC), and to reveal the role and mechanism of epidermal growth factor receptor(EGFR) in the pathogenesis of NPC. NPC cell line CNE2 cells were cultured in serum-free medium and stimulated by transforming growth factor-? (TGF-?) for 24 h in experimental group. Control CNE2 cells were cultured at the same condition but without TGF-? stimulation. The culture medium of control and experimental cells was desalted and concentrated through ultrafiltration to prepared the total secreted proteins. Two-dimensional gel electrophoresis (2-DE) was used to separate the secreted proteins of control and experimental cells, PDQuest software was applied to analyze 2-DE images, and the differential protein spots between the control and experimental cells were identified by desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The 2-DE patterns of the secreted proteins of TGF-? stimulated and un-stimulated CNE2 cells were established, 22 differential proteins spots between the two groups of cells were found, and 8 non-redundant proteins were identified with MALDI-TOF-MS, the functions of which were involved in invasion, metastasis, apoptosis and proliferation of cancer cells. The data will be valuable for further to study the role and mechanism of EGFR in the pathogenesis of NPC.
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Objective To study the expression and activation of signal transducer and activator of transcription 3(STAT3) in human nasopharyngeal carcinoma (NPC) tissues and its relation with the expression of latent membrane protein 1(LMP1) encoded by Epstein-Barr virus.Methods The expressions of LMP1,STAT3 and phosphated STAT3(p-STAT3) in 45 cases of NPC and 27 cases of chronic nasopharyngitis were studied by immunohistochemical method.Correlation between protein expressions was analyzed.Results The positive rates of LMP1,STAT3 and p-STAT3 in NPC were significantly higher than those in chronic nasopharyngitis(P0.05).Conclusion There are overexpression and abnormal activation of STAT3 protein in NPC tissues.LMP1 may play a role in the activation of STAT3.
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Objective To study the value of CT perfusion in differential diagnosis of local recur and local radiofibrosis of nasopharyngeal carcinoma(NPC) at pre-and post-radiotherapy.Methods Dynamic CT scan was performed in 71 objectives: 14 in local recur group,22 in radiofibrosis group,15 in pre-radiotherapy NPC group and 20 in control group,the time-density curve(TDC) and the data of perfusion parameters were recorded and analyzed statistically.Results The TDC feature between local recur group,pre-radiotherapy NPC group and radiofibrosis group,control group had difference.The average of blood flow(BF) had significant differences by two by two comparison in four groups(P
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Objective To improve the cognition of abnormal changes of the medial soft tissue of styloid process (MSTSP) in nasopharyngeal carcinoma after radiotherapy. Methods CT scans of nasopharynx in 39 cases with NPC that had change at the MSTSP in NPC after radiotherapy were performed.CT findings of recurrence and non-recurrence at MSTSP were studied and compared carefully.Results In the recurrence cases, the MSTSP was thickening and compactness in all cases, in the non-recurrence cases ,only 37.04% (10/27) were thickening and compactness at MSTSP. In all the cases which had thickening and compactness of the MSTSP, the borders were slightly protruding in 9 cases and the borders were straight in 3 cases in the recurrence cases, while the borders were straight in 7 cases and the borders were slightly hollow in 3 cases,and there wasn't any case appeared as protruding of borders in the non-recurrence cases. All the cases were dealt with statistic methods and had remarkable difference between them(P
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One hundred and thirty five patients with carcinoma of the nasopharyx were treated by radiation therapy in the Department of Radiation Oncology, Yonsei Cancer Center, Yonsei University between August 1977 and July 1987. Of the 30 patients omitted: 8 had distant metastases at initial diagnosis or during radiotherapy; 18 patients refused or did not received a full course of radiation therapy, and four had not been confirmed histologically. The remaining 105 patients were analyzed to determine the incidence and patter of distant metastases. Diagnosis of distant metastases was made based on clinical signs and radiography, even though histologic confirmation was not made. Twenty-six patients developed distant metastases after definite irradiation of nasopharyx and neck, an incidence rate of 24.8%. The common sites of distant metastases were, in descending order, bone, lung, liver, and brain. There was a strong correlation between Ho's N stage and distant metastases rate. But sex, age, histologic subtype (squamous cell and undifferentiated cell), AJC T and N stage, treatment modalities (radiotherapy alone and radiotherapy combined with chemotherapy) were not significant. Of those patients who developed distant metastases, 80.8% were discovered within 2 years of their radical radiotherapy. The prognosis for nasopharyngeal carcinoma patients developing distant metastases was poor: median survival was nine months and 80% of those patients died within two years of the initial diagnosis of distant metastasis.