Rare esophageal carcinoma-primary adenoid cystic carcinoma of the esophagus: A case report

BACKGROUND Esophageal adenoid cystic carcinoma(EACC)is an exceedingly rare malignant tumor of the esophagus,posing significant challenges in the clinic.CASE SUMMARY This report detailed the case of a 72-year-old male whose diagnosis of EACC was confirmed through postoperative histopathological examination.The patient underwent thoracoscopy-assisted radical resection of the esophageal tumor,coupled with lymph node dissection.Pathological findings revealed an adenoid cystic carcinoma infiltrating the entire layer of the muscularis propria,locally extending into the outer membrane of the esophageal fiber,involving the cardia and exhibiting no lymph node metastasis.The patient’s condition was classified as primary EACC,T3N0M0,per the American Joint Committee on Cancer(2017;8th edition).One month after surgery,the patient received postoperative adjuvant radiation therapy.CONCLUSION In addressing the rarity and high potential for biopsy misdiagnosis of EACC,this study delved into its diagnostic methods and treatment.

Novel milestones for early esophageal carcinoma:From bench to bed

Esophageal cancer(EC)is the seventh most common cancer worldwide,and esophageal squamous cell carcinoma(ESCC)accounts for the majority of cases of EC.To effectively diagnose and treat ESCC and improve patient prognosis,timely diagnosis in the initial phase of the illness is necessary.This article offers a detailed summary of the latest advancements and emerging technologies in the timely identification of ECs.Molecular biology and epigenetics approaches involve the use of molecular mechanisms combined with fluorescence quanti-tative polymerase chain reaction(qPCR),high-throughput sequencing technology(next-generation sequencing),and digital PCR technology to study endogenous or exogenous biomolecular changes in the human body and provide a decision-making basis for the diagnosis,treatment,and prognosis of diseases.The invest-igation of the microbiome is a swiftly progressing area in human cancer research,and microorganisms with complex functions are potential components of the tumor microenvironment.The intratumoral microbiota was also found to be connected to tumor progression.The application of endoscopy as a crucial technique for the early identification of ESCC has been essential,and with ongoing advancements in technology,endoscopy has continuously improved.With the advancement of artificial intelligence(AI)technology,the utilization of AI in the detection of gastrointestinal tumors has become increasingly prevalent.The implementation of AI can effectively resolve the discrepancies among observers,improve the detection rate,assist in predicting the depth of invasion and differentiation status,guide the pericancerous margins,and aid in a more accurate diagnosis of ESCC.

Genes Expression Profile Difference in Peripheral Blood Between Esophageal Carcinoma Patients and Normal Subjects

Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early conceration associated genes. Methods： The total RNA was extracted and purified in the peripheral blood obtained from the patients with esophageal carcinoma and normal subjects. The first strand of cDNA was synthesized through retro-transcription and labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with a piece of 4096 double dot human whole gene chip. The acquired image was analyzed by microarrav suite software using a digital computer, and the intensity of ttuorescence signal and its ratio were calculated. Results： A total of 92 genes were screened out and its expression difference was more than 2 times in the peripheral blood between the patients with esophageal carcinoma and normal subjects. Among these, the expression difference of 36 genes was more than 3 times. Two human urokinase plasminogen activator surface receptor （UPAR） genes, 80K-L protein gene, human protein tyrosine-phosphatase gent arid proto-oncogene protein mRNA were significantly up-regulated, while the collagen V type （α-2 gene was markedly down-regulated. Conclusion： 80K-L protein gene, tyrosinephophatase gene, proto-oncogene protein arid the collagen V type α-2 gene might be associated with the ontogenesis, development and its metastasis in the esophageal carcinoma. The UPAR gene may play important roles in the diagnosing the micrometastasis in the peripheral blood of esophageal carcinoma.

Clinical observation of nedaplatin concurrent with radiotherapy for mid-advanced nasopharyngeal carcinoma and esophageal carcinoma

Objective: The aim of our study was to evaluate the short-term efficacy and the toxic reaction of nedaplatin concurrent with radiotherapy for mid-advanced nasopharyngeal carcinoma and esophageal carcinoma. Methods: Thirty-four patients were confirmed diagnosis with cancer by pathologic results. All patients were given 6MV X-ray for radiotherapy, Dt 66-70 Gy/33-35 f/6-7 w, concurrently administrated nedaplatin (30 mg/m2) once a week (6 times). Results: A total 34 patients were enrolled, of whom 33 patients were available for objective response, 1 patient of esophageal cancer quit for allergic reaction. The response rate (RR) of nedaplatin-contained therapy for nasopharyngeal carcinoma and esophageal carcinoma were 90.0% and 76.9%, respectively. The major toxic reaction was bone marrow suppression observed in 25 patients (73.5%), in which grade III aleukocytosis was observed in 3 patients (8.8%), grade III + IV thrombocytopenia in 3 patients (8.8%). And 6 patients (17.6%) showed gastrointestinal tract reaction. There were 4 patients with radiation esophagitis in the 13 patients with esophageal carcinoma. Conclusion: Nedaplatin can increase the therapeutic effect of radiation. Its incidence rate of bone marrow suppression is high, but the gastrointestinal tract reaction and renal toxicity is low and mild.

Clinical features, outcomes and treatment-related pneumonitis in elderly patients with esophageal carcinoma

AIM: To investigate the clinical features and prognoses of elderly patients with esophageal carcinoma and to compare the effects of radiotherapy and rates of treatment-related pneumonitis (TRP) between elderly and non-elderly patients.

Expression of Livin and vascular endothelial growth factor in different clinical stages of human esophageal carcinoma

AIM： To investigate the role of Livin and vascular endothelial growth factor （VEGF） in human esophageal carcinoma, and analyze its relationship to clinical stages.METHODS： Expression of Livin in fresh esophageal cancer tissues was detected by immunohistochemistry （IHC）, Western blotting and reverse transcriptasepolyrnerase chain reaction （RT-PCR）, and VEGF by Western blotting and RT-PCR. All statistical analyses were performed by SPSS version 11.0. RESULTS： Livin positivity was also significantly correlated with tumor stages, increasing with tumor progression. Expression of Livin and VEGF increased with the process of esophageal carcinoma. In the fourth clinical stage, expression of Livin and VEGF was the most significant. Expression of Livin was positively correlated with VEGF. CONCLUSION： Over-expression of Livin and VEGF contributes to the pathogenesis of esophageal carcinoma.

Neoadjuvant chemoradiotherapy for resectable esophageal carcinoma:A meta-analysis

AIM:To compare neoadjuvant chemoradiotherapy and surgery with surgery alone for resectable esophageal carcinoma. METHODS:We used MEDLINE and EMBASE databases to identify eligible studies and manual searches were done to ensure no studies were missed.Trial validity assessment was performed and a trial quality score was assigned. RESULTS:Eleven randomized controlled trials(RCTs) including 1308 patients were selected.Neoadjuvant chemoradiotherapy significantly improved the overall survival compared with surgery alone.Odds ratio(OR) [95%confidence interval(CI),P value],expressed as neoadjuvant chemoradiotherapy and surgery vs surgery alone,was 1.28(1.01-1.64,P=0.05)for 1-year survival,1.78(1.20-2.66,P=0.004)for 3-year survival,and 1.46(1.07-1.99,P=0.02)for 5-year survival.Postoperative mortality increased in patients treated by neoadjuvant chemoradiotherapy(OR: 1.68,95%CI:1.03-2.73,P=0.04),but incidence of postoperative complications was similar in two groups (OR:1.14,95%CI:0.88-1.49,P=0.32).Neoadjuvant chemoradiotherapy lowered the local-regional cancer recurrence(OR:0.64,95%CI:0.41-0.99,P=0.04), but incidence of distant cancer recurrence was similar (OR:0.94,95%CI:0.68-1.31,P=0.73).Histological subgroup analysis indicated that esophageal squamous cell carcinoma did not benefit from neoadjuvantchemoradiotherapy,OR(95%CI,P value)was 1.16(0.85-1.57,P=0.34)for 1-year survival,1.34 (0.98-1.82,P=0.07)for 3-year survival and 1.41 (0.98-2.02,P=0.06)for 5-year survival. CONCLUSION:Neoadjuvant chemoradiotherapy can raise the survival rate of patients with esophageal adenocarcinoma.

Expression and significance of heat shock protein 70 and glucose-regulated protein 94 in human esophageal carcinoma

AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70 and grp94 in 78 human esophageai cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis. RESULTS: Both esophageai cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageai carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8% (56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageai cancer and adjacent normal tissues (P<0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P<0.01). CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70 is related to the differentiation of esophageai carcinoma.

RELATIONSHIP BETWEEN LYMPH NODE METASTASES IN ESOPHAGEAL CARCINOMA AND ITS PROGNOSIS

Objective: To study the relationship between lymph node metastases in esophageal carcinoma and its prognosis. Methods: We obtained 1500 resected lymph nodes from the specimen of 86 patients with resected esophageal carcinoma and checked these lymph nodes by routine histopathology. Additiionally, frozen tissue sections of 540 lymph nodes classified as tumor-free by routine histopathology were screened for micrometastases by immunohistology with the monoclonal antibody Ber-EP4. Results: Forty-two patients (49%) had pN0 disease, and 61 patients (71%) had lymph node micrometastases detected by immunohistochemistry, skip metastases detected by routine histopathology were present in 26%(11/42) of pN0 and 41%(18/44) of pN1 patients. Skipping of micrometastases detected by immunohistochemistry was found in 71%(61/86). Twenty-six of 42 patients (62%) with tumor staged as pN0 and 35 of 44 patients (80%) with stage pN1 had nodal micrometastasis. The presence of micrometastases was associated with a significantly decreased relapse-free time and overall survival (P<0.0001 and P=0.004, respectively). Conclusion: Lymph node skip metastases are a frequent event in esophageal carcinoma. Extensive lymph node sampling, in conjunction with immunohistochemical detection, will lead to accurate staging and prognosis.

Anti-tumor Effect and Its Mechanisms of Ursolic Acid on Human Esophageal Carcinoma Cell Eca-109 in Vivo

Objective： To investigate the anti-tumor effect and possible mechanisms of ursolic acid on human esophageal carcinoma in vivo. Methods： Atransplanted tumor model by injecting Eca-109 cells into subcutaneous tissue of BALB/c nude mice was established. 40 nude mice bearing tumors were randomly divided into 4 groups and 0.2 ml saline or 0.2 ml ursolic acid （25-100 mg·kg^-1·d^-1） was injected into abdominal cavity respectively once everyday and lasted for fourteen days. The changes of tumor volume were measured continuously and tumor inhibition rate was calculated. The morphological changes of apoptosis were observed by electron microscope. The expressions of COX-2, bcl-2 and Bax protein in transplanted tumors were detected by immunohistochemistry. At last the PGE2 level of transplanted tumors was detected byradioimmunoassay. Results： Treatment of nude mice with 25, 50, or 100 mg·kg^-1·d^-1 of ursolic acid significantly inhibited the growth of the human esophageal carcinoma tumor in nude mice and induced Eca-109 cells apoptosis as demonstrated by electron microscopy analyses. The expressions of COX-2 and bcl-2 in the transplanted tumors were decreased in ursolic acid groups, while the Bax increased. The PGE2 level of transplanted tumors was decreased in ursolic acid groups with adose-relatedmanner. Conclusion： Ursolic acid has anti-tumor effects against human esophageal carcinoma cells in vivo, which are likely mediated via induction of tumor cell apoptosis and inhibition of COX-2 and PGE2.

Atrial fibrillation after surgery for esophageal carcinoma:Clinical and prognostic significance

AIM： To retrospectively evaluate the clinical relevance, perioperative risk factors, outcome of different pharmacological prophylaxis, and short-term prognostic value of atrial fibrillation （AF） after surgery for esophageal carcinoma. METHODS： We retrospectively studied 63 patients with AF after surgery for esophageal carcinoma in comparison with 126 patients without AF after esophagectomy during the same time. Postoperative AF incidence was related to different clinical factors possibly involved in its occurrence and short-term survival. RESULTS： A strong relationship was observed between AF and postoperative hypoxia, history of chronic obstructive pulmonary disease （COPD）, postoperative thoracic-gastric dilatation, age older than 65 years, male sex and history of cardiac disease. No difference was observed between the two groups with regard to shortterm mortality and length of hospital stay. CONCLUSIONS： AF occurs more frequently after esophagectomy in aged and male patients. Other factors contributing to postoperative AF are history of COPD and cardiac disease, postoperative hypoxia and thoracicgastric dilatation.

Value of endoscopic methylene blue and Lugol's iodine double staining and detection of GST-n and telomerase in the early diagnosis of esophageal carcinoma

AIM：To explore the expressions of GST-π and telomerase activity in esophageal carcinoma and premalignant lesions and to investigate the value of endoscopic methylene blue （MB） and Lugol＇s iodine double staining. METHODS： Seventy-two patients with esophagopathy were sprayed endoscopically with MB and Lugol＇s iodine in proper order and the areas stained blue and brown, and the area between the blue and brown stains were obtained. Depending on the pattern of mucosal staining, biopsy specimen was obtained. GST-π and telomerase activity in specimens were examined by immunohistochemistry and PCR-based silver staining telomeric repeat amplification protocol, respectively. RESULTS： After MB and Lugol＇s iodine staining, the area between both the colors was obtained in 64 of the 72 patients and the areas were stained blue and brown in all of the 72 patients. Association test of two simultaneous ordinal categorical data showed a correlation between the esophageal mucosal staining and the esophageal histology （P〈0.005）. The expression of GST-π and telomerase activity in esophageal carcinoma and premalignant lesions increased. The expression of GST-π and telomerase activity in dysplasia and carcinoma was significantly higher than that in normal epithelium （P〈0.005）. The expression in hyperplasia was slightly higher than that in normal epithelium. With the lesions progressing from low- to moderate- to high-grade dysplasia, the positive rate increased （P〈0.025）. Expression of GST-π was correlated with that of telomerase activity in dysplasia and carcinoma （φ= 0.4831, P〈0.005;φ= 0.3031, P〈0.025, respectively）; but there was no correlation between them in normal epithelium and hyperplasia. CONCLUSION： The expression of GST-π and telomerase may be an early event in the carcinogenesis of esophagus. They may play an induced and synergistic role with each other in the carcinogenesis of esophagus. Endoscopic MB and Lugol＇s iodine double staining and detection of GST-π and telomerase activity may contribute to the early diagnosis of esophageal carcinoma.

Clinical application of metallic stents in treatment of esophageal carcinoma

AIM: To evaluate the effects of self-expanding metal stents (SEMS) in patients with malignant esophageal obstruction and to analyze their prognosis and complications. METHODS: Seventy-four metallic stents were placed under fluoroscopic guidance in 66 patients with esophageal obstruction secondary to carcinoma, of whom, 6 cases were complicated by fistula. RESULTS: After seventy-two stents were successfully used in 66 cases without any severe complications (technical successful rate was 97%), the dysphagia score improved from 3.3±0.6 to 0.8±0.5 (P<0.01), and life quality improved significantly in all these patients. All fistulae were sealed immediately after coated stents were inserted in the six patients. New stents were placed in two patients: the stent migrated more than 2 cm, in one patients and the stent slipped into stomach in the other. Minor bleeding was found only in 28 patients during the operation. Reobstruction was found in 12 patients, but was successfully cured under endoscopy. The survival rate was 78%, 57% and 11% for 6 mo, 1 year and 2 years respectively. CONCLUSION: Placement of SEMS is a simple, safe, quick and efficient surgical method for treating esophageal carcinoma obstruction. It may be used mainly as a palliative treatment of esophageal obstruction secondary to carcinoma.

Effects of Oxymatrine on the Apoptosis of Human Esophageal Carcinoma Eca109 Cell Line and Its Mechanism

The effects of Oxymatrine （Oxy） on the proliferation and apoptosis of human esophageal carcinoma Ecal09 cell line and the mechanism were investigated. The human esophageal carcinoma Eca 109 celis were cultured in vitro. The Oxy-induced apoptosis of Eca 109 cells was assayed by using flow cytometry. The expressions of p-ERKII2, Cyclin D1, p21^waf/cipl, Bax and Bcl-2 were detected by Western blot. Flow cytometry revealed that Oxy could induce the apoptosis of Eca l09 cells. Western blot showed that Oxy of different concentrations suppressed the expressions of p-ERK1/2, Cyclin D1 and Bcl-2, but up-regulated the expression of p21waf/cip1 and Bax, and the ratio of Bax/Bcl-2 was increased. It was suggested the Oxy could induce the apoptosis of Eca l09 cells, which might be related to the upregulation of p21waf/cip1 and the downregulation of p-ERK1/2, Cyclin D1 and p21^waf/cip1. The possible pathway may be related to Bcl-2/Bax.

Serum transforming growth factor-β1 level reflects disease status in patients with esophageal carcinoma after radiotherapy

AIM: To evaluate the relationship between changes in serum transforming growth factor β1 (TGFβ1) level and curative effect of radiotherapy (RT) in patients with esophageal carcinoma. METHODS: Ninety patients with histologically confi rmed esophageal carcinoma were enrolled. Serum samples for TGFβ1 analysis were obtained before and at the end of RT. An enzyme-linked immunosorbent assay was used to measure serum TGFβ1 level. Multivariate analysis was performed to investigate the relationship between disease status and changes in serum TGFβ1 level. RESULTS: Serum TGFβ1 level in patients with esophageal carcinoma before RT was signifi cantly higher than that in healthy controls (P < 0.001). At the end of RT, serum TGFβ1 level was decreased in 67.82% (59/87) of the patients. The overall survival rate at 1, 3 and 5 years was 48.28% (42/87), 19.54% (17/87) and 12.64% (11/87), respectively. Main causes of death were local failure and regional lymph node metastasis. In patients whose serum TGFβ1 level decreased after RT, the survival rate at 1, 3 and 5 years was 61.02% (36/59), 28.81% (17/59) and 18.64% (11/59), respectively. The survival rate at 1 year was 17.86% (5/28) in patients whose serum TGFβ1 level increased after RT, and all died within 18 mo (P < 0.01). CONCLUSION: Serum TGFβ1 level may be a useful marker for monitoring disease status after RT in patients with esophageal carcinoma.

Expression level of beta-catenin is associated with prognosis of esophageal carcinoma

AIM： To examine the association of beta-catenin with the clinicopathologic features and prognosis of esophageal squamous cell carcinoma （ESCC）. METHODS： Beta-catenin mRNA expression level in 40 ESCC patients （28 males and 12 females, age range 38-82 years, median 60 years） was analyzed by real- time PCR. Beta-catenin mRNA expression levels in tumor cells were categorized as weaker （level 1） or equal to or stronger （level 2） than those in endothelial cells. We examined the correlation between the beta-catenin expression and the clinicopathological factors and prognosis of ESCC patients. RESULTS： Level 2 beta-catenin expression was found in 29 patients. ESCC with level 2 expression had a higher rate of lymphnode metastasis （0.0776±0.0369 vs 0.3413±0.1803, P 〈 0.001） and deeper tumor invasion （0.0751±0.0356 vs 0.3667±0.1928, P 〈 0.001）, and a poorer survival rate （P = 0.0024） than ESCC with level I expression. CONCLUSION： Beta-catenin expression in ESCC is of great importance.

Expression of miR-1304 in patients with esophageal carcinoma and risk factors for recurrence

BACKGROUND Esophageal carcinoma is a malignant gastrointestinal tumor with a very poor prognosis.MicroRNA(miR)-1304 is a newly discovered non-coding RNA,which shows differential expression in other cancers,and its clinical value in esophageal carcinoma remains unclear.AIM To explore the expression of miR-1304 in patients with esophageal carcinoma and its clinical value.METHODS The expression of miR-1304 in patients with esophageal carcinoma was analyzed based on the data on miR in esophageal carcinoma downloaded from The Cancer Genome Atlas database.Quantitative real-time polymerase chain reaction was adopted to determine the expression of miR-1304 in the tissues and serum of patients.The clinical diagnostic value of miR-1304 and independent factors for recurrence and prognosis of esophageal carcinoma were then analyzed.The potential target genes of miR-1304 were predicted,and then analyzed based on gene ontology,Kyoto Encyclopedia of Genes,and Genomes,and protein-protein interaction.RESULTS The expression of miR-1304 in the tissues and serum of patients with esophageal carcinoma increased,and was also increased according to the database.Patients with high expression of miR-1304 suffered increased rates of tumor≥3 cm,low differentiation and stage II+III.miR-1304 had a diagnostic value in identifying esophageal carcinoma,tumor size,differentiation and TNM stage.Tumor size,differentiation,TNM stage,and miR-1304 were independent risk factors for recurrence of esophageal carcinoma,and they had certain predictive and diagnostic value for the recurrence of esophageal carcinoma.Seventy-eight patients showed a 3-year survival rate of 38.46%,and patients with high expression of miR-1304 had a relatively lower survival rate.Multivariate analysis revealed that tumor size,differentiation,recurrence and miR-1304 were independent factors for the prognosis of patients.MiRTarBase,miRDB,and Targetscan predicted 20 target genes in total.Gene ontology enrichment analysis found 18 functions with aP<0.05,and Kyoto Encyclopedia of Genes,and Genomes analysis found 11 signal pathways with aP<0.05.String analysis of protein co-expression found 269 relationship pairs,of which co-expression with epidermal growth factor was the most common.CONCLUSION miR-1304 can be used as a potential indicator for the diagnosis and recurrence of esophageal carcinoma and for survival of patients with this disease.

Increased N-terminal pro-brain natriuretic peptide level predicts atrial fibrillation after surgery for esophageal carcinoma

AIM: To evaluate the value of plasma N-terminal pro- brain natriuretic peptide (NT-proBNP) level for predicting postoperative atrial fibrillation (AF) in patients undergoing surgery for esophageal carcinoma. METHODS: NT-proBNP levels were measured in 142 patients 24 h before and 1 h after surgery for esophageal carcinoma. All patients having a preoperative cardiac diagnosis by electrocardiogram (ECG), remained under continuous monitoring for at least 48 h after surgery, and then underwent clinical cardiac evaluation until discharge. RESULTS: Postoperative AF occurred in 11 patients (7.7%). AF patients were significantly older (69.6 ± 12.2 years vs 63.4 ± 13.3 years, P = 0.031) than non-AF patients. There were no significant differences in history of diabetes mellitus, sex distribution, surgical approach, anastomosis site, intraoperative hypotension and postoperative fever. The preoperative plasma NT-proBNP level was significantly higher in patients who developed postoperative AF (121.3 ± 18.3 pg/mL vs 396.1 ± 42.6 pg/mL, P = 0.016). After adjustment for age, gender, chronic obstructive pulmonary disease (COPD), history of cardiac diseases, hypertension, postoperative hypoxia and thoracic-gastric dilation, NT-proBNP levels were found to be associated with the highest risk factor for postoperative AF (odds ratio = 4.711, 95% CI = 1.212 to 7.644, P = 0.008).CONCLUSION: An elevated perioperative plasma BNP level is a strong and independent predictor of postoperative AF in patients undergoing surgery for esophageal carcinoma. This finding has important implications for identifying patients at higher risk of postoperative AF who should be considered for preventive antiarrhythmic therapy.

In vivo anti-tumor effect of hybrid vaccine of dendritic cells and esophageal carcinoma cells on esophageal carcinoma cell line 109 in mice with severe combined immune deficiency

AIM： To develop a fusion vaccine of esophageal carcinoma cells and dendritic cells （DC） and observe its protective and therapeutic effect against esophageal carcinoma cell line 109 （EC109）. METHODS： The fusion vaccine was produced by fusing traditional polyethyleneglycol （PEG）, inducing cytokine, sorting CD34＋ magnetic microbead marker and magnetic cell system （MACS）. The liver, spleen and lung were pathologically tested after injection of the fusion vaccine. To study the therapeutic and protective effect of the fusion vaccine against tumor EC109, mice were divided immune group and therapeutic group. The immune group was divided into P, E, D and ED subgroups, immunized by phosphate buffered solution （PBS）, inactivated EC109, DC and the fusion vaccine respectively, and attacked by EC109 cells. The tumor size, weight, latent period and mouse survival period were recorded and statistically analyzed. The therapeutic group was divided into four subgroups： P, inactivated EC109, D and ED subgroups, which were attacked by EC109 and then treated with PBS, inactivated EC109, DC, and EC109-DC respectively. Pathology and flow cytometry were also used to study the therapeutic effect of the fusion vaccine against EC109 cells.RESULTS： Flow cytometry showed that the expression of folate receptor （FR）, EC109 （C）, Des （D） in human nasopharyngeal carcinoma cell line （HNE1） （B） was 78.21%, 89.50%, and 0.18%, respectively. The fusion cells （C） were highly expressed. No tumor was found in the spleen, lung and liver after injection of the fusion vaccine. Human IgG was tested in peripheral blood lymphocytes （PBL）. In the immune group, the latent period was longer in EC109-DC subgroup than in other subgroups, while the tumor size and weight were also smaller than those in ED subgroup. In the therapeutic group, the tumor size and weight were smaller in ED subgroup than in P, inactivated EC109 and DC subgroups. CONCLUSION： Fusion cells are highly expressed not only in FR but also in CD80. The fusion vaccine has a distinctive protective effect against tumor EC109 and can inhibit the growth of tumor in mice, and its immune protection against tumor attack is more significant.

Integration of human papillomavirus 18 DNA in esophageal carcinoma 109 cells

AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.