Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells

In order to study the mechanism of the effect of heparin on apoptosis in carcinoma cells, the nasopharyngeal carcinoma cell line CNE2 was used to identify the effect of heparin on apoptosis associated with the expression of c-myc, bax, bcl-2 proteins by use of Hoechst 33258 staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), agarose gel electrophoresis, and flow cytometry, as well as Western blot analysis. The results showed that heparin induced apoptosis of CNE2 cells including the morphologic changes such as reduction in the volume, and the nuclear chromatin condensation, as well as the 'ladder pattern' revealed by agarose gel electrophoresis of DNA in a concentration-dependent manner. The number of TUNEL-positive cells was dramatically increased to 33.6+/-1.2% from 2.8+/-0.3% by treatment with heparin in different concentrations (10 to approximately 40 kU/L). The apoptotic index was increased to 32.5% from 3.5% by detecting SubG1 peaks on flow cytometry. Western blot analysis showed that levels of bcl-2, bax and c-myc were significantly overexpressed by treatment with the increase of heparin concentrations. These results suggest that heparin induces apoptosis of CNE2 cells, which may be regulated by differential expression of apoptosis-related genes.

The initial results of Epstein-Barr virus (EBV)-encoded latent membrane protein-1 (LMP-1) for screening nasopharyngeal carcinoma (NPC)

Objective: Early diagnosis of nasopharyngeal carcinoma (NPC) is an important method to improve the survival rate.However,the sensitivity and specificity of the screening protocols which was widely used in clinic now are considered to be unsatisfactory.Epstein-Barr virus (EBV)-encoded latent membrane protein-1 (LMP-1) is one of the proteins that have been suggested to be a classic oncogene with transformation properties.The current study set out to discuss the clinical significance of LMP-1 on the screening of NPC.Methods: Three hundred patients who visited our institution (Department of Radiation Oncology,Fujian Provincial Cancer Hospital,Fuzhou,China) with ENT symptoms between 2007 and 2008 were involved in this study,and all of them were agreed to be involved in this investigation.Not only did they undergo nasopharyngeal swab to obtain cells for the LMP-1 polymerase chain reaction (PCR) analysis,but also nasopharyngeal biopsy were taken to identify the diagnosis.Results: An amount of DNA that was sufficient for PCR was extracted from 243 (81%) swab samples,the positive rate of LMP-1 of those with non-nasopharyngeal carcinoma was 3.85% (4/108),which was much lower than those with nasopharyngeal carcinoma (P < 0.05).By detecting LMP-1 in nasopharyngeal swabs,NPC was diagnosed with a sensitivity of 88.15% (119 of 135 patients),specificity of 96.30% (104 of 108 patients),a positive predictive value of 95.2% (119 of 123 patients),a negative predictive value of 86.67% (104 of 120 patients),accuracy of 91.77%,and Youden index of 84.45%.Conclusion: The nasopharyngeal swab coupled with PCR-based EBV LMP-1 detection have high sensitivity and specificity,and also good repeatability,it could serve as part of the screening program for high-risk populations.

THE IMPACT OF RADIOTHERAPY COURSE LENGTH ON THE TREATMENT RESULTS OF NASOPHARYNGEAL CARCINOMA(NPC)

Analyses made among four radiotherapy schedules for NPC in order to determine whether there is an impact of radiotherapy course length on treatment results.A series of 320 NPC patients were divided into four radiation treatment branches each with a schedule, this clinical trial was non-randomized.Radiotherapy course length factor was considered with a derivative LQ model formula that biological effective dose(BED)=nd[1 +d/(α/β)]-(T-28).The four branches were:1.split-course 103 cases, with an intermediary rest of 3-4 weeks,mean total dose 70Gy/35fx,73d, BED 51.6 Gy; 2.continuous 115 cases, 72Gy/36fx, 61d, BED 62.6 Gy;3.hyperfractionation I 52 cases, 1.5 Gy b.i.d.,time interval(Ti)≥6 hr, 75Gy/49fx,57d,BED 65.5Gy;4. hyperfractionation Ⅱ 50 cases, 1.2 Gy b.i.d., Ti≥6hr,76Gy/60fx,59d,BED 63.0 Gy.Treatment results were compared with 1-, and 3-year loco-regional recurrent rates,and 1-,and 3-year survival rates, and these rates were of a negative interrelation with prolonged course duration, but of a positive one with BED valas.Continuous branch was of a course mean 12 days shorter than the spilt-course one, its treatment results were nearly 10% higher in some subgroups;and hyperfractionation branches were slightly better than continuous one.

CONDITIONED MEDIUM OF HUMAN NASOPHARYNGEAL CARCINOMA EPITHELIOID CELL LINE CNE_(1) CONTAINED THE ACTIVITIES OF TRANSFORMED GROWTH-INHIBITING FACTORS

The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that the SFCM stimulated the growth of long-term serum-free cultured CNE4 cells in ac-cordence with the fact that the growth rate of long-term serum-free cultured CNE1 cells was directly proportional to the plating density. Alternatively 5% SFCM inhibited the growth of short-term serum-free cultured CNE4 cells by 51% in which the indicator cell remained the responsiveness state of growing in the serum-supplemented medium to the effector of interest. Furthermore, SFCM resulted in the inhibition of anchorage-independent growth of CNE4 cells and A431 cells. Also in soft agar test. SFCM reduced the colony formation of NRK(?),9F cells in the presence of EGF or EGF plus TGF-β. These finding suggested that CNE4 secreted autocrine growth stimulating factor(s) and growth inhibiting factor(s) in the serum-free medium, the latter strongly reverse malignant phenotypes of CNE4 and A431 cells in serum-supplemented surrounding.

Biomarkers for enhancing the radiosensitivity of nasopharyngeal carcinoma

Nasopharyngeal carcinoma(NPC) is a common head and neck malignancy. The incidence of NPC is higher in Southern China and Southeast Asia compared with Western countries. Given its high radiosensitivity, the standard treatment for NPC is radiotherapy. However, radioresistance remains a serious obstacle to successful treatment. Radioresistance can cause local recurrence and distant metastases in some patients after treatment by radiation. Thus, special emphasis has been given to the discovery of effective radiosensitizers. This review aims to discuss the biomarkers, classified according to the main mechanisms of radiosensitization, which can enhance the sensitivity of NPC cells to ionizing radiation.

Familial nasopharyngeal carcinomas possess distinguished clinical characteristics in southern China

Objective： To compare clinical characteristics between familial nasopharyngeal carcinomas（NPCs） and sporadic NPCs in Guangdong province, China, a high-risk area.Methods： Between 1991 and 2001, 993 NPC patients treated at the Cancer Center of Sun Yat-Sen University in Guangdong were randomly selected as probands. Information about NPC among the probands＇ relatives and other information were obtained from a retrospective review of the patients＇ medical records. The patients were divided into sporadic NPC, low-frequency familial NPC（one NPC patient in addition to the proband in three generations）, and high-frequency familial NPC（2 or more additional NPC patients in three generations） groups. Pathological and clinical characteristics were compared among these groups.Results： Of the 993 patients, 131（13.2%） had a familial history of NPC. The average age at diagnosis was the lowest in the high-frequency familial NPC group（39 years; P=0.048）. Although the overall survival（OS）, distant metastasis-free survival（DMFS）, and disease-free survival（DFS） rates did not differ between familial and sporadic NPCs, the locoregional recurrence-free survival（LRFS） rate increased in the order sporadic NPCs, low-frequency familial NPCs, and high-frequency familial NPCs（P=0.009）, with 5-year rates of 70%, 83%, and 87%, respectively. Multivariate analysis showed that family history of NPC was an independent favorable prognostic factor for LRFS, with adjusted hazard ratio（a HR） of 0.548, 95% CI（0.342-0.878）. The high LRFS for familial NPCs was mainly noted among young, advanced-stage patients who received continuous radiation treatment.Conclusions： Genetic factors may play an important role in the etiology of high-frequency familial NPC and underlie the early age of onset and sensitivity to radiotherapy.

Plasma microRNA profiling in nasopharyngeal carcinoma patients reveals miR-548q and miR-483-5p as potential biomarkers

MicroRNAs(miRNAs), which play a role in tumorigenesis, may also serve as diagnostic or prognostic biomarkers. However, studies on human miRNA profiles in plasma from nasopharyngeal carcinoma(NPC) patients are in their infancy. Here, we used microarrays to perform systematic profiling of human miRNAs in plasma from NPC patients. We subsequently used real-time quantitative polymerase chain reaction(Q-PCR) to validate miRNAs with aberrant expression that could serve as potential biomarkers. By comparing the plasma miRNA profiles of 31 NPC patients and 19 controls, 39 of 887 human miRNAs were found to be aberrantly expressed. Considering the fold change and P value, miR-548q and miR-483-5p were validated in 132 samples from 82 NPC patients and 50 controls. Moreover, high expression of miR-548q and miR-483-5p was further found in 3 NPC cell lines and clinical biopsy tissues from 54 NPC patients and 22 controls. Our results revealed that miR-548q and miR-483-5p are potential biomarkers of NPC. Combining the receiver operating characteristic(ROC) analyses of these 2 miRNAs, an area under the ROC curve(AUC) of 0.737 with 67.1% sensitivity and 68.0% specificity were obtained, showing the preliminary diagnostic value of plasma miRNAs. Moreover, most NPC patients with a poor outcome exhibited high expression(> median) of miR-548q(70.6%) and miR-483-5p(64.7%) in tissue samples, indicating their prognostic value. The high expression levels of miR-548q and miR-483-5p in plasma, cell lines, and clinical tissues of NPC patients indicate that their roles in NPC should be explored in the future.

Retrospective analysis of results of treatment for nasopharyngeal carcinoma in Macao

Background： Nasopharyngeal carcinoma （NPC） is a common malignancy in Southeast Asia, however, a full consensus has not yet been reached as to the value of comprehensive treatment for NPC. This study was designed to evaluate the epidemiological characteristics of NPC and their prognostic value, as well as the long-term efficacy of NPC treatment. Patients and methods： A total of 248 patients, with different stages of NPC, were included in this study. Results： The 5-year overall survival （OS） rates for patients in stages I, II, lII and IV were 90.48%, 76.71%, 76.89% and 33.87%, respectively （P=0.000）, while the respective 5-year progression-free survival （PFS） rates were 85.15%, 72.36%, 63.88% and 26.26% （P=0.000）. The respective 5-year OS rates, according to stage, for the group that received radiotherapy combined with chemotherapy and for the group that received radiotherapy only were as follows： stages I and II, 81.67% and 79.59% （P=0.753）; stage III, 79.91% and 70.38% （P=0.143）; stage IV,, 35.22% and 0% （P=0.000）. The respective 5-year PFS rates in these groups were as follows： stages I and II, 75.83% and 74.98% （P=0.814）; stage III, 74.08% and 42.25% （P=0.027）; stage IV,, 27.31% and 0% （P=0.000）. Conclusions： Clinical staging appears to be the most important prognostic factor for NPC. As the stage number increases, both the 5-year OS and PFS significantly decrease. Adding chemotherapy to radiotherapy was not advantageous for patients with stage I or II NPC, however the addition of chemotherapy to radiotherapy significantly improved OS and PFS in patients with stage IV NPC. The addition of chemotherapy improved PFS, but not OS in patients with stage III NPC.

The clinical effects of concurrent radiochemotherapy in cases suffered from stages III to IVa nasopharyngeal carcinoma

Objective： The aim of the study was to investigate the clinical effects of concurrent radiochemotherapy in treating locally advanced （Stages III-IVa） nasopharyngeal carcinomas （NPCs）. Methods： A total of 95 patients who suffered from nasopharyngeal carcinoma （Stages III-IVa） was divided into two groups： Group concurrent radiochemotherapy （Group CCRT, n = 49） and Group radiotherapy （Group RT, n = 46）. The two groups were both delivered conventional fractionated radiotherapy, while Group CCRT was delivered three cycles chemotherapy of PF （DDP ＋ 5-Fu） regimen or PLF （DDP ＋ 5-Fu ＋ CF） regimen. Results： The complete remission rate and total remission rate of Group CCRT were higher than those of Group RT, and the differences were of statistical importance （X2 = 4.72-7.19, P 〈 0.05）. The one-year overall survival （OS） rate was calculated by life table method, in Group CCRT, it was higher than that of Group RT and the difference was of statistical importance （X2 = 4.24, P 〈 0.05）. The 3-year overall survival （OS） rate, nasopharyngeal control rate and cervical lymph nodes＇ control rate of Group CCRT were all higher than those of Group RT and the differences were of statistical importance （X2 = 4.28-4.40, P 〈 0.05）. The 5-year overall survival （OS） rate and metastasis-free rate of Group CCRT were higher than those of Group RT and the differences were of statistical importance （X2 = 3.96-8.26, P 〈 0.05）. The incidence rates of acute toxicities in Group CCRT were obviously higher than those in Group RT, and the difference of gastrointestinal reaction was of statistical importance （X2 = 11.70, P 〈 0.05）. Conclusion： This study has demonstrated that concurrent radiochemotherapy can improve the remission rate, overall survival rate and locally control rate. The toxicities of concurrent radiochemotherapy can be tolerated by the patients.

The Research Progress of Long Noncoding RNAs in Nasopharyngeal Carcinoma

Long noncoding RNA (lncRNA) is a leader of the degree of more than 200 nucleotides, almost do not have the function of protein coding endogenous RNA molecules. Recent studies show that, lncRNA is not encoded protein, but it has a wide range of biological functions, and lncRNA in human diseases, especially in oncology, more and more attention has been paid to its role. Nasopharyngeal carcinoma (NPC) is a common malignant tumor of the head and neck in South China, which poses a serious threat to people’s health and life. Studies found that lncRNA is widely involved in the invasion, metastasis and prognosis of nasopharyngeal carcinoma (NPC). In this article, we will review the research progress of lncRNA in nasopharyngeal carcinoma.

Radiation therapy concurrent with weekly cisplatin therapy for loco-regionally advanced nasopharyngeal carcinoma:outcomes of a clinical trial

Objective:The purpose of this study was to define the maximum tolerated dose(MTD) by describing the doselimiting toxicity(DLT) of weekly cisplatin concurrently with conventional plus 3-dimensional conformal radiotherapy(CT + 3DCRT) in patients with loco-regionally advanced nasopharyngeal carcinoma(NPC).Methods:Patients with loco-regionally advanced NPC(III/IVa stage) were enrolled into a dose-escalating study.Toxicity was graded according to Common Terminology Criteria for Adverse Events version 3.0(CTCAE v3.0).MTD was defined when 2 of 6 patients developed DLT.The starting dose of cisplatin was 15 mg/m2/w,with a subsequent dose escalation of 5 mg/m2/w in cohorts of 3 new patients.CT + 3DCRT was given to the nasopharynx and the upper neck;the lower neck was treated by a single anterior field irradiation.The prescription dose was 70-80 Gy by 35-40 fractions to the nasopharynx gross tumor,and 66-70 Gy by 33-35 fractions to the positive neck lymph nodes.Results:From Jun.2008 to Sep.2009,24 patients received complete chemoradiotherapy,and all of them were eligible for toxicity evaluation.On the first five dose levels of 15 mg/m2/w and 35 mg/m2/w,no patient experienced DLT.On the next dose level of 40 mg/m2/w,1 patient experienced DLT of grade 3 myelosuppression for 1.4 weeks,and among the additional 3 patients,no one developed DLT.On the seventh dose level of 45 mg/m2/w,all the patients developed grade 3 myelosuppression for more than 1 weeks,and the dose-escalating trial stopped.The 23(95.8%) patients achieved clinical complete remission(CR) in the local site;22(91.7%) achieved CR in the regional site,and all patients got CR 3 months later.After a median follow-up of 16.4 months,1 patient developed liver metastases 2 months later,1 patient developed bone metastases 10 months later and 22 kept disease-free survival.Conclusion:The MTD of cisplatin weekly with concurrent CT + 3DCRT for local advanced NPC is 40 mg/m2/w,with myelosuppression as DLT.

Yi Qi Jie Du Decoction Inhibits Proliferation and Induces Apoptosis of Nasopharyngeal Carcinoma Stem Cells Through Mitochondrial Apoptosis Pathway

Objective To assess the effects of Yi Qi Jie Du Decoction(YQJDD)on nasopharyngeal carcinoma stem cells(NPC-SCs)and investigate the underlying mechanism.Methods NPC-SCs were collected in serum-free culture system and identified by the sphere formation assay.The effect of YQJDD on the proliferation of NPC-SCs was detected using the Cell Counting Kit-8 assay.The change of mitochondrial membrane potential(ΔΨm)in NPC-SCs following treatment with YQJDD was investigated by JC-1 staining.The levels of apoptosisassociated proteins were examined using western blot analysis.Results YQJDD significantly inhibited the proliferation of NPCSCs in dose-and time-dependent manners.JC-1 staining revealed that YQJDD lowered theΔΨm of NPC-SCs in a timedependent manner.After treatment with YQJDD,the expression of cleaved caspase-3,-7 and-9,cleaved poly-ADP ribose polymerase,P21 and P53 were increased,while the expression of Survivin in NPC-SCs was decreased.However,the expression of cleaved caspase-8 remained nearly unchanged.Conclusions YQJDD inhibits the growth and induces apoptosis of NPC-SCs via an intrinsic apoptosis pathway.

Detection of Survivin mRNA in nasopharyngeal carcinoma by real-time fluorescence quantitative RT-PCR

Objective： To establish the method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in nasopharyngeat carcinoma （NPC） tissues. Methods： The total RNA was extracted from NPC cell line CNE-2 and tissues with Trizol and then been transcribed reversely to cDNA, a method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in NPC tissues had been established, in which chronic nasopharyn-gitis patients＇ nasopharynx tissues treated as control group. Results： The expression of Survivin mRNA all could be detected either in CNE-2 cells, NPC tissues or in chronic nasopharyngitis patients＇ nasopharynx tissues, and there was higher the expression level of Survivin mRNA in NPC tissues than which in chronic nasopharyngitis patients＇ nasopharynx tissues, the difference was significant （P 〈 0.01）. The expression of Survivin mRNA could be detected both in stage Ⅰ ＋ Ⅱ and stage Ⅲ ＋ Ⅳ NPC, and there was no significant difference in relative quantifications of gene expression between these two groups （P 〉 0.05）. There was no relationship between Survivin mRNA expression and age and sex of NPC patients （P 〉 0.05）. Conclusion： Real time fluorescence quantitative RT-PCR is a rapid, effective and high sensitive method for detecting the expression of Survivin mRNA in NPC tissues. The overexpression of Survivin mRNA may play some roles in pathogenesis of NPC.

Expressions of genes related to genome stability and DNA repair in nasopharyngeal carcinoma clustering families

Objective： The aim of the study was to observe the expressions of genes related to genome stability and DNA repair in the members of nasopharyngeal carcinoma （NPC） clustedng families. Methods： In the Zhongshan City where there is highly incidence rate of NPC, we chose the members of the NPC clustering families as objects, and the patients of nasopharyngitis and NPC as the control group. We isolated the RNA from the nasopharyngeal tissue, and synthesized its cRNA, the genome stability and DNA repair genes chip technique, chemiluminescent detection and real-time fluorescence quantita- tive technique were used to examine the genome stability and DNA repair genes in the nasopharyngeal tissue. Results： More genome stability and DNA repair genes were up-regulated in the members of the NPC clustering families than the NPC patients, and the range of up-regulated was high, with the over up-regulated 100 times genes including TEP1, MSH4, PMS2LI. Fewer genome stability and DNA repair genes were down-regulated in the members of the NPC clustering families than the NPC patients, the ubiquitin genes almost were down-regulated, the results also could be confirmed by real-time fluorescence quantitative PCR. Conclusion： There are specially expression character of genome stability and DNA repair genes in the members of NPC clustering families.

THE RELATIONSHIP BETWEEN THE INVASION OF PARAPHARYNGEAL SPACES AND THE INVOLVEMENT OF CERVICAL NODES AND POSTERIOR GROUP CRANIAL NERVES IN NASOPHARYNGEAL CARCINOMA

Two hundred cates of nasopharyngeal carcinoma (NPC) admitted to this department from Feb. 1985 to May. 1988 were analysed according to the CT scanning and clinical findings of the primary lesions prior to radiotherapy. The results showed that involvement of parapharyngeal space was very common in NPC, about 80% (160/200 cases) ; particularly unilateral or bilateral retro-styloid spaces, about 69.5% (139/200 cases). It was proposed that patients with NPC had a high Incidence of ipsilateral cervical node metastasis. Contralateral cervical node metastasis was rare. The development of cervical node metastasto in NPC has two modes: one Is direct Infiltration of the retro-stylold space by the lesion; the other Is along the nasopharyngeal lymphatic rete. The data also showed that patients with NPC who presented symptoms of Ⅸ- Ⅲ cranial nerve paralyses always had ipsilateral or bilateral retro- styloid space Infiltrations.

Clinical role of ^(18)F-FDG PET/CT-based simultaneous modulated accelerated radiotherapy treatment planning for locally advanced nasopharyngeal carcinoma

Objective The aim of this study was to compare the long-term local control, overall survival, and late toxicities of positron emission tomography/computed tomography （PET/CT）-guided dose escalation radio- therapy versus conventional radiotherapy in the concurrent chemoradiotherapy treatment of locally ad- vanced nasopharyngeal carcinoma （NPC）. Methods Atotal of 48 patients with stage IIl-IVa NPC were recruited and randomly administered PET/CT- guided dose escalation chemoradiotherapy （group A） or conventional chemoradiotherapy （group B）. The dose-escalation radiotherapy was performed using the simultaneous modulated accelerated radiotherapy technique at prescribed doses of 77 gray （Gy） in 32 fractions （f） to the gross target volume （GTV）： planning target volume （PTV） 1 received 64 Gy/32 f, while PTV2 received 54.4 Gy/32 f. Patients in group B received uniform-dose intensity-modulated radiotherapy, PTV1 received 70 Gy/35 f and PTV2 received 58 Gy/29 f. Concurrent chemotherapy consisted of cisplatin [20 mg/m2 intravenous （IV） on days 1-4] and docetaxel （75 mg/m2 IV on days 1 and 8） administered during treatment weeks 1 and 4. All patients received 2-4 cycles of adjuvant chemotherapy of the same dose and drug regimen. Results The use of fluorine-18-fluorodeoxyglucose （18F-FDG） PET/CT significantly reduced the treat- ment volume delineation of the GTV in 83.3% （20/24） of patients. The 5-year local recurrence-free survival rates of the two groups were 100% and 79.2%, respectively （P = 0.019）. The 5-year disease free survival （DFS） rates were 95.8% and 75.0%, respectively （P = 0.018）. The 5-year local progression-free survival and DFS rates were significantly different. The 5-year overall survival （OS） rates were 95.8% and 79.2%, re- spectively. Differences in OS improvement were insignificant （P = 0.079）. Late toxicities were similar in the two groups. The most common late toxicities of the two arms were grade 1-2 skin dystrophy, xerostomia, subcutaneous fibrosis, and hearing loss. There were no cases of grade 4 late toxicity. Conclusion The use of 18F-FDG PET/CT-guided dose escalation radiotherapy is well tolerated and can reduce local recurrence rates for patients with locally advanced NPC compared to conventional chemora- diotherapy.

Induction of cytotoxic T lymphocyte respones in vivo after immunotherapy with dendritic cells in patients with nasopharyngeal carcinoma

The aim of the present study was to determine the efficacy of immunotherapy with dendritic cells to elicit EBV-specific CTL-immunity in advanced cases of EBV-positive patients with nasopharyngeal carcinoma （NPC） and to determine the safety and toxicity of this preparation. Nine cases of histologically confirmed patients with NPC undergoing treatment with radiological therapy were enrolled in this study. Dendritic cells, generated in vitro from blood monocytes of patients were cultured and matured with cytokines and then infected with recombinant adenovims vaccine containing EBV-latent membrane protein-2 （Ad-LMP2）. On 9 days＇ cultivation of cells, the matured DCs were harvested, irradiated with 60^Co and then injected intradermally to patients with NPC. The injections were performed 3 times totally. After immunization, the CTL responses were assayed by means of cytotoxicity and epitope-specific IFN-γ production. The results of this trial showed that all patients could tolerate this kind of treatment without any side effect, during which marked increase of LMP2-specific CIL-responses could be demonstrated in 5 patients of this group. And the level of IgA/VCA antibody decreased in 8 of 9 patients, thus accounting for a better prognosis for these patients. All patients will be followed up for another one year. At least, the present work shows that intradermal vaccination with autologons DCs infected with recombinant Ad-LMP2 adenovirus is a safe procedure in NPC patients, in which this procedure can enhance the LMP2-specific CTL responses in patients. These data are encouraging to develop more effective vaccine strategies for the treatment of nasopharyngeal carcinoma.

ASSOCIATION OF pIgR POLYMORPHISMS WITH NASOPHARYNGEAL CARCINOMA

Objective： Epstein-Barr virus （EBV） associated nasopharyngeal carcinoma （NPC） is an important squamous cell cancer endemic in southern China and Southeast Asia. pIgR （polymeric immunoglobulin receptor） gene plays central roles during immune responses and EBV inflammatory and therefore is a good candidate susceptibility gene for NPC. This study is to evaluated potential associations between pIgR gene and NPC susceptibility. Methods： Sequencing was used to identify multiple single nucleotide polymorphisms （SNPs） within the exon regions of pIgR in Guangdong population. Four SNPs were genotyped in 528 NPC patients and 408 normal individuals to perform case-control study. Results： There was no statistical difference in the allele frequencies of each SNP （P〉0.05）. After categorized into 2 groups by age of 45 y, in the group of age below 45 the minor allele T frequency of C888oT was 7%, whereas 4% in controls, with significant difference （P〈0.05）. The Odds Ratio （OR=1.84） also showed higher risk of NPC with individuals carried the minor alleles. Conclusion： The result has proved that SNP C8880T is associated with NPC susceptibility and pIgR gene might play a certain role in oncogenesis and development of NPC.

Local definitive intensity-modulated radiation therapy recommended for patients initially diagnosed with nasopharyngeal carcinoma with distant metastasis after an effective systemic chemotherapy

Objective The aim of the study was to propose a hypothesis that local definitive intensity-modulated radiation therapy(IMRT) should be recommended for initially diagnosed metastatic nasopharyngeal carcinoma(NPC) and demonstrate its feasibility.Methods Recently published papers on local definitive radiotherapy for initially diagnosed metastatic NPC were reviewed to propose a hypothesis.Results Several studies revealed the survival benefits of adding local definitive radiotherapy to the systemic chemotherapy in patients initially diagnosed with metastatic NPC.Conclusion We suggested that local definitive IMRT should be recommended in patients initially diagnosed with NPC with distant metastasis after an effective systemic chemotherapy, which may possibly prolong their survival time and potentially treat the disease.

Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method

Latent mermbrane protein 1(LMP1)is known as an oncoprotein in nasopharyngeal carcinoma(NPC)cells,which is considered to have a strong association with growth,invasion and metastasis of NPC cells through lipid rafts.Met hy-A-cyclodextrin(M3CD)can disrupt lipid rafts through cholesterol depletion.In this study,we revealed that MICD induced apoptosis in two kinds of NPC cells including CNE1 cells,a LMP1 negative nasopharyngeal squamous carcinoma cell line,and CNE1-LMP1 cells,a LMP1-positive nasopharyngeal squamous carcinoma cell line.Furthermore,the impact of MBCD on LMP1 was investigated by fuorescence resonance energy transfer(FRET)method in NPC cells.Synchronized tempo spatial and spectral detection of LMP1/LMP1 interaction were performed using fluorescence microscope and spectrograph.FRET efficiency indicated that LMP1/LMP1 interaction gradully enhanced after M9CD treatment.MTT assays showed that M8CD caused strong cytotoxicity in CNE1 cells,but cauused relatively weaker cytotoxicity in CNE1-LMP1 cells,which indicated that LMP1 may regulate sensitivity of NPC cells to MBCD.Then,detection of cleaved caspase-3 in two kinds of NPC cells indicated that LMP1 may inhibit activation of caspase 3.These results strongly suggested that MBCD can induce apoptosis of NPC cells,but enhancing of LMP1/LMP1 interaction may likely resist apoptosis through inhibiting cleavage of caspase-3.