Expression of Coxsackievirus and Adenovirus Receptor in Human Lung Cancer： Possible Clinical Significance

OBJECTIVE To explore the relationship between CAR and the development of human lung cancer, as well as to provide the basis for the clinical treatment of lung cancer using an adenovirus vector-based gene therapy. METHODS CAR expression was assessed immunohisto- chemically in tumoral, paraneoplastic and normal samples from 112 lung cancer patients. At the same time, the mRNA and protein expression of CAR in 32 cases were determined by RT-PCR and Western blot. The relationship between CAR expression and clinicopathologic parameters was statistically analyzed. RESULTS There was no expression of CAR in normal lung tissue but a little in paraneoplastic tissue. The positive rate was 43% in squamous cell carcinoma, and 70% in adenocarcinoma. Both were much significantly higher than that in paraneoplastic tissue. The CAR expression level in adenocarcinoma was higher than that in squamous cell cancer, mRNA expression by RT-PCR and protein expression by Western blot were consistent with immunohistochemistry results. CONCLUSION CAR is overexpressed in human lung cancer, especially in adenocarcinoma. This data offer the reliable basis for adenovirus-mediated gene therapy of lung cancer; more important, CAR may take part in the formation or development of lung cancer; this may be exploitable for the development of antibody-directed therapy in human lung cancer.

Inhibitory Effect of Coxsackie Adenovirus Receptor on Invasion and Metastasis Phenotype of Ovarian Cancer Cell Line SKOV3

Full-length coxsackie adenovirus receptor (CAR) eukaryotic expression plasmid was transfected into an ovarian cell line, SKOV3, and its effect on the change of malignant metastasis phenotype was explored. CAR mRNA and protein expression levels among 4 ovarian cancer cell lines (A2780, SKOV3, SW626, CAOV3) and the positive control 293 (a transformed human embryo kidney cell line) was detected by using semi-quantitative RT-RCR and Western blot and compared. CAR-negative SKOV3 was transfected with the eukaryotic expression plasmid containing a full-length CAR cDNA and mock-vector respectively. The positive clones were screened by G418. The biological behavior changes of positive transfected cells were gauged by colony formation in soft agar assay and cell adhesion assay. Among the cell lines, there were obviously different CAR expression levels. CAR could not be detected in SKOV3. In transfected cell group, CAR expression was enhanced obviously as compared with non-transfected or mock-transfected groups. Cell adhesion in the transfected group was promoted. The number of colony formation was reduced significantly in transfected groups (25.32±8.91) as compared with that in non-transfected group (88.75±13.98) and mock-transfected group (82.53±19.37). Among the 4 ovarian cancer cell lines, CAR expression level was variable. Exogenous CAR expression had a potential role in inhibiting the malignant metastasis phenotype of ovary cancer cells.

Expression of coxsackie and adenovirus receptor in small cell lung cancer

Objective: We explored the expression of coxsackie and adenovirus receptor (CAR) in small cell lung cancer (SCLC) tissue. Methods: CAR expression in 31 SCLC was assessed in formaldehyde-fixed, paraffin-embedded tissue according to the EnVision immunohistochemistry procedure, while 3 samples of surgical specimens of non-malignant lung disease were taken as the negative control. Results: We observed that the expression of CAR was detectable positive in all the 31 cases from the small cell lung cancer tissue, in contrasting that non-malignant lung tissue control. Conclusion: The high expression of CAR appeared in SCLC tissue indicates that it play an important role in of adenovirus vector-based gene therapy in SCLC.

Expression and Significance of Coxsackie and Adenovirus Receptor in Renal-Cell Carcinoma

OBJECTIVE To investigate the expression of Coxsackie and Adenovirus receptor （CAR） in renal-cell carcinoma and the relationship of the CAR to the biological behavior of the carcinomas. METHODS The immunohistochemical SP method was used to detect the expression of Coxsackie and Adenovirus receptor in 48 cases of renal- cell carcinoma and in 12 cases of normal renal tissue 2 cm away from the tumor tissue. RESULTS The positive rates of CAR were 100% in 12 cases of para-tumor normal renal tissue and 35.4% in 48 cases of renal-cell carcinoma respectively. The difference of CAR expression between them was significant （P〈0.05）. The grades of the tumor were as follows： 22 in Grade Ⅰ, 17 in Grade Ⅱ and 9 in Grade Ⅲ with the CAR positive rate being 54.5%, 23.5% and 11.1%, respectively. There was a negative correlation between CAR expression and tumor grading （P〈0.05）. In addition, the number of the cases in stages I to IV were 19, 13, 11 and 5 respectively, with the respective positive rates being 57.9%, 30.8%, 18.2% and 0.0%, i.e. there also was a negative relationship between CAR expression and the stage （P〈0.05）. CONCLUSION CAR expression is down-regulated in renal-cell carcinoma compared with normal tissue. The level of CAR may be a sensitive predictor of differentiation, invasion and metastasis. Loss of CAR expression correlates with the invasive phenotype in our analysis of renal-cell carcinoma.

Down-regulation of Coxsakie and Adenovirus Receptor during Embryo Implantation

In this study,real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression.Both localization and quantity were evaluated in the uteri ob-tained at days post coitus (dpc) 2.5,4.5,6.5,8.5.Outcome of PCR was assessed by 2-ΔΔCt method.Im-age Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohisto-chemical sections.We found relatively weak CAR expression in the mouse uteri during implantation window.PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference).At dpc 8.5,CAR expression was increased slightly again.It is concluded that during implantation,the expression of CAR mRNA and protein is declined,resulting in the impairment of tight junction between cavity epithelium cells.After implantation window closure,CAR appears again to maintain epithelium stability.CAR might play an important role during embryo implantation procedure.

Expression of Coxsackie and Adenovirurus Receptor and its Significance in Human Lung Cancer

OBJECTIVE To study the relationship between the coxsackie and adenovirus receptor （CAR） and the development of human lung cancer. To optimize adenovirus vector-based gene therapy.METHODS The expression of CAR in 112 cases of lung cancer was examined using immunohistochemistry. At the same time, the relationship between CAR expression and clinicopathologic characteristics was analyzed,RESULTS ：lhere is a little expression of CAR in normal lung tissue. Compared with paraneoplastic epithelial tissue of the lung, the expression of CAR is generally up-regulated in tumor tissues showing a significant dif- ference （P〈0.01）. The positive rate of CAR expression in squamous cell carcinoma was 43.1%, and in adenocarcinoma 70.2%, with the difference between the two rates being statistically significant （P〈0.01）. Compared to the paraneoplastic tissues, the difference in CAR positive expression was 35.4% for squamous cell carcinoma and 38.3% for adenocarcinoma. But the difference in different stages of squamous cell carcinoma had no statistical significance （P〉0.05）. However, the expression of CAR was at a high level in the bronchioalveolar carcinomas as 80.4% were CAR positive. This research showed that there was a specially high expression of CAR in adenocarcinomas.CONCLUSION CAR is expressed in human lungs at a low level and up-regulated in the tumor tissues, suggesting that there is a relationship between adenocarcinoma and CAR. This research provides a basis for planning a regimen of gene therapy using an adenovirus vector,

Correlations among Persistent Viral Infection,Heart Function and Chinese Medicine Syndromes in Dilated Cardiomyopathy Patients

Objective：To investigate the correlations among persistent viral infection,heart function and Chinese medicine（CM） difined-syndromes in patients with dilated cardiomyopathy（DCM）.Methods：Fifty patients with DCM in the First Affiliated Hospital of Zhejiang Chinese Medical University from October 2009 to December 2011 were selected as the research subjects,and 30 healthy people were simultaneously selected as the normal control group to detect persistent viral infections after admission.The CM syndrome type and grade of heart function were then evaluated.The expression level of Coxsackie adenovirus receptor（CAR） was detected using the flow cytometry（FCM） technique,coxsackie virus RNA（CVB-RNA） using reverse transcription polymerase chain reaction（RTPCR）,and the plasma brain natriuretic peptide（BNP） level with a Triage meter plus diagnosis instrument.Finally,the parameters such as left ventricular end diastolic diameter（LVEDd） and left ventricular ejection fraction（LVEF） were measured by ultrasonic cardiogram.Person correlation analysis was used for measured data,Spearman correlation analysis for rating data,and the Chi-square test for numerical data.Results：CVB-RNA was positive in 22 patients（44%） with DCM,while only 6 cases（20%） were CVB-RNA-positive in the normal control group,with a significant difference between the two groups（P〈0.01）.The expression level of CAR was significantly elevated in the DCM group compared with the normal control group（P〈0.01）.In CVB-RNA-positive patients（22 cases）,the expression level of CAR was significantly higher than in CVB-RNA-negative patients（28 cases;P〈0.01）.In the DCM patients,there was a positive correlation between the CAR expression and the BNP level（r=0.34,P〈0.05）,while no significant difference was found between the CAR expression and the LVEF and LVEDd（r=-0.32,0.30,P〉0.05）.There was no clear correlation between virus infection and the CM syndrome types in DCM patients（r=-0.22,P〉0.05）.According to the sequence of syndrome types：phlegm → qi deficiency → blood stasis → hydroretention with asthenic yang（from low to high）,a positive correlation was existed between the BNP levels and CM syndrome types（r=0.139,P〈0.05）.Conclusion：The expression of CAR on the surface of white cells could be used to detect persistent viral infection.The expression level of CAR and heart function in DCM patients were highly correlated.The expression level of BNP may serve as an objective index for differentiating CM syndromes for patients with DCM.