Association of chemokine and chemokine receptor expression with the invasion and metastasis of lung carcinoma.

The chemokine system has been reported to be utilized and manipulated by tumor cells in order to promote local tumor growth and distant dissemination. The present study aimed to investigate the expression of three chemokine ligand-receptor axes in lung carcinoma tissues. Tumor and healthy normal tissue samples were obtained from 120 lung carcinoma patients following surgical resection. Immunohistochemistry and reverse transcription quantitative polymerase chain reaction were used in order to identify the protein and messenger (m)RNA expression of chemokines, including chemokine (C-X-C motif) ligand (CXCL)12/stromal cell-derived factor (SDF)-1, CXCL8/interleukin (IL)-8, chemokine (C-C motif) ligand (CCL)19 and CCL21, and the corresponding chemokine receptors, chemokine (C-X-C motif) receptor (CXCR)4, CXCR1, CXCR2 and chemokine (C-C motif) receptor (CCR)7, respectively. The results revealed that compared with the normal lung tissues, lung carcinoma tissues expressed significantly higher mRNA levels of CXCL12/SDF-1, CXCR4, CXCL8/IL-8, CXCR2, CCL19 and CCR7 (P<0.01). In four histological subtypes, adenocarcinoma presented dominant expression of CXCR4, CXCR2, CXCL8/IL-8 and CCL19 (P<0.05). In addition, it was demonstrated that tumor staging was inversely correlated with chemokine receptor CCR7 and CXCR2 mRNA expression as well as positively correlated with CXCL12/SDF-1, CXCL8/IL-8 and CCL19 mRNA levels (P<0.05). Lymph node metastasis presented a positive correlation with CXCR4, CXCR2 and CXCL8/IL-8 expression and a negative correlation with CCL19 and CCR7 expression (P<0.05). Furthermore, vascular invasion was more prevalent in patients with higher expression levels of CXCR4, CCR7 or CCL19 (P<0.01). In conclusion, these data suggested that the ligand-receptor interaction of CXCL8-CXCR2, CXCL12-CXCR4 and CCL19-CCR7 may be involved in the tumorigenesis of lung carcinoma. Higher expression levels of chemokines and lower expression of chemokine receptors indicated poor tumor staging. The CXC chemokine receptors, CXCR4 and CXCR2, promoted lymphatic metastasis through the activation of their specific ligands, while CCL19 and its receptor CCR7 had an essential role in hematogenous metastasis of lung carcinoma.

One case of inflammatory myofibroblastic tumor--a case report.

Inflammatory myofibroblastic tumor is a rare mesenchymal tumor, it can also be found on the trunk, head and neck, internal organs, and soft tissue. It has been named as inflammatory pseudotumor, plasma cell granuloma, solitary mast cell tumor, pseudotumor pneumonia and tissue cells, and other inflammatory pseudotumor. The main treatment of inflammatory myofibroblastic tumor patients is through surgical complete excision of the lesion.

Overexpression of Pygopus-2 is required for canonical Wnt activation in human lung cancer.

Lung cancer is the most common cause of cancer-related mortality worldwide. It is necessary to improve the understanding of the molecular mechanisms involved in lung cancer in order to develop more effective therapeutics for the treatment of this fatal disease. The canonical Wnt signaling pathway has been known to be important in a number of cancer types, including lung cancer. Pygopus (Pygo) is a recently identified downstream component of the Wnt signaling pathway required for ß-catenin/T-cell factor (TCF)-dependent transcription. However, the role of Pygo in lung cancer remains to be elucidated. The present study showed that Pygo2 is overexpressed in human lung cancer tissue samples and cell lines. Expression levels of Pygo2 were found to correlate with cytosolic ß-catenin protein levels in the samples examined. Co-immunofluorescent staining showed that Pygo2 protein accumulated in the nuclei and colocalized with nuclear ß-catenin in lung cancer cell lines expressing Pygo2. To investigate the functional importance of the Pygo2 overexpression in lung cancer, short hairpin RNA (shRNA) was used to knockdown Pygo2 mRNA in lung cancer cells expressing the gene. Pygo2 shRNA was observed to inhibit cell proliferation and decrease ß-catenin/TCF-dependent transcriptional activity in vitro. Furthermore, Pygo2 shRNA significantly suppressed lung cancer xenograft models in vivo (P<0.05). These results suggested that Pygo2 is a putative therapeutic target for human lung cancer and overexpression of Pygo2 may be important for aberrant Wnt activation in lung cancer.

[Autopsy clinicopathologic comparison in different age groups of patients with coronary atherosclerosis].

OBJECTIVE: To explore the features of coronary atherosclerosis among the patients at different ages and examine their risk factors and pathological characteristics. METHODS: Retrospective analysis was performed for the clinicopathologic data of 87 patients with coronary atherosclerosis confirmed through autopsy from February 1986 to December 2011 at our hospital. They were divided into 3 groups according to age (20 - < 40 years, n = 12; 40 - < 60 years, n = 24; ≥ 60 years, n = 51). Comparative analysis was performed for the degree of coronary artery stenosis and cardiac pathological changes, vulnerable plaque occurrence and morphological features of acute coronary syndrome among different patient groups. RESULTS: There were 56 males and 31 females with a mean age of 66.8 years (range: 23 - 80). Great statistical differences existed in the degree of coronary artery stenosis between different genders (P < 0.05) while there was no significant difference among the above age groups (P > 0.05). In the above age groups of patients with acute coronary syndrome, the incidence of big lipid core was 23.5%, 38.5% and 53.8% (χ(2) = 6.282, P = 0.043), that of thin fibrous cap 29.4%, 41.0% and 58.8% (χ(2) = 6.589, P = 0.037), that of inflammatory cell in 2 filtration 58.8%, 69.2% and 85.0% (χ(2) = 7.435, P = 0.024) and that of calcification formation 35.3%, 56.4% and 71.3% respectively (χ(2) = 8.599, P = 0.014). And the incidence of vulnerable plaque occurrence was 76.5%, 84.6% and 88.8% (χ(2) = 1.850, P = 0.397) in the above age groups. CONCLUSIONS: The features and risk factors of coronary atherosclerosis are different according to different ages and genders. Thus we should pay more attention to the early diagnosis and reasonable treatments of young patients and those with both mild coronary stenosis and vulnerable plaque.

[Application of combined telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis].

OBJECTIVE: To evaluate the application of traditional cytomorphology, telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis of pleural effusion and bronchoalveolar lavage samples. METHODS: A total of 123 agar-paraffin double-embedded pleural effusion and bronchoalveolar lavage fluid samples were enrolled into study. The cytomorphologic features were reviewed and correlated with immunocytochemical findings and telomerase activity. RESULTS: Telomerase activity was detected in 53 specimens using the real-time telomeric repeat amplification protocol. Amongst the cases studied, 39 samples (31.7%) contained overtly malignant cells while 20 cases (16.0%) were equivocal by conventional cytology. After verification by immunocytochemistry and clinical follow-up data, the diagnostic accuracy of telomerase activity and cytology was 87.0% and 82.1%, respectively. The sensitivity (97.6%) and specificity (100.0%) of cytology examination, when combined with telomerase activity analysis, were greater than those of cytology examination or telomerase activity analysis alone. CONCLUSIONS: Telomerase activity analysis can be used as an adjunctive investigative tool in cytology assessment of pleural effusion and bronchoalveolar lavage samples. The diagnostic accuracy can be further improved with the application of immunocytochemistry on agar-paraffin double-embedded cell block tissues.

Frizzled-8 as a putative therapeutic target in human lung cancer.

Lung cancer is the leading cause of cancer related deaths worldwide. It is necessary to better understand the molecular mechanisms involved in lung cancer in order to develop more effective therapeutics for the treatment of this disease. Recent reports have shown that Wnt signaling pathway is important in a number of cancer types including lung cancer. However, the role of Frizzled-8 (Fzd-8), one of the Frizzled family of receptors for the Wnt ligands, in lung cancer still remains to be elucidated. Here in this study we showed that Fzd-8 was over-expressed in human lung cancer tissue samples and cell lines. To investigate the functional importance of the Fzd-8 over-expression in lung cancer, we used shRNA to knock down Fzd-8 mRNA in lung cancer cells expressing the gene. We observed that Fzd-8 shRNA inhibited cell proliferation along with decreased activity of Wnt pathway in vitro, and also significantly suppressed A549 xenograft model in vivo (p<0.05). Furthermore, we found that knocking down Fzd-8 by shRNA sensitized the lung cancer cells to chemotherapy Taxotere. These data suggest that Fzd-8 is a putative therapeutic target for human lung cancer and over-expression of Fzd-8 may be important for aberrant Wnt activation in lung cancer.

[The clinicopathologic analysis in autopsy patients with cardiovascular disease].

OBJECTIVE: To investigate the clinical autopsy results of patients died of cardiovascular disease or other disease complicated with cardiac damage. METHODS: Complete autopsy was performed on 86 cases with uncertain cause of death. Through integrating clinical diagnosis and treatment with gross autopsy findings and microscopic observations, 86 autopsies were determined the major cause of death. RESULTS: In 86 autopsies, 69 cases were heart disease. Differences between pathological diagnosis and clinical diagnosis were compared. Twenty-seven cases were cardiac deaths, with diagnosis accordance rate of 81.5%. Forty-two cases died of non-cardiac disease but complicated with heart disease or involving the heart which accelerated the death in patients, with accordance rate of 78.6%. CONCLUSION: Scientific and correct performance of autopsy was important to determine the causes of death, to promote development of related disciplines and to improve the diagnosis and treatment of the diseases.

Use of serum circulating CCNB2 in cancer surveillance.

Cyclin B2 (CCNB2), a member of the cyclin protein family, has been found to be up-regulated in human cancers. To evaluate the potential use of circulating CCNB2 in serum in cancer surveillance, we examined relative expression levels of serum circulating CCNB2 mRNA in 103 cancer patients, 19 normal controls, and 40 benign disease patients using real-time quantitative reverse transcriptase polymerase chain reaction. We found that the relative expression level of circulating CCNB2 mRNA in cancer patients was significantly higher (p<0.0001) than that in normal controls and benign diseases group. Circulating CCNB2 mRNA level was significantly (p<0.001) correlated with cancer stage and metastasis status. Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.87 and 0.83 (p<0.05) in identifying cancer patients' metastasis status in lung and digestive tract cancer, respectively. Moreover, we observed that expression levels of circulating CCNB2 mRNA in cancer patients significantly decreased (p=0.0084) after their therapeutic treatments. These data suggest that detection of serum circulating CCNB2 mRNA may have potential clinical applications in screening and monitoring of metastasis and therapeutic treatments.

Use of serum circulating CCNB2 in cancer surveillance.

Cyclin B2 (CCNB2), a member of the cyclin protein family, has been found to be up-regulated in human cancers. To evaluate the potential use of circulating CCNB2 in serum in cancer surveillance, we examined relative expression levels of serum circulating CCNB2 mRNA in 103 cancer patients, 19 normal controls, and 40 benign disease patients using real-time quantitative reverse transcriptase polymerase chain reaction. We found that the relative expression level of circulating CCNB2 mRNA in cancer patients was significantly higher (p<0.0001) than that in normal controls and benign diseases group. Circulating CCNB2 mRNA level was significantly (p<0.001) correlated with cancer stage and metastasis status. Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.87 and 0.83 (p<0.05) in identifying cancer patients' metastasis status in lung and digestive tract cancer, respectively. Moreover, we observed that expression levels of circulating CCNB2 mRNA in cancer patients significantly decreased (p=0.0084) after their therapeutic treatments. These data suggest that detection of serum circulating CCNB2 mRNA may have potential clinical applications in screening and monitoring of metastasis and therapeutic treatments.

Detection of telomerase activity in cultured cells and tumor tissue of lung carcinoma by modified telomeric repeat amplification protocol.

Telomerase activity is found in various cell types including stem cells, neoplastic cells, and immortalized cells, suggesting a close association with their proliferation capacity. The telomeric repeat amplification protocol (TRAP) has been traditionally used to detect semi-quantitatively the telomerase activity by polyacrylamide gel electrophoresis (PAGE), which is difficult to apply for large scale analysis because of laborious post-PCR manipulation and potential carryover contamination. In the present study, a specific reverse primer was designed and the TRAP protocol was adapted to either PAGE or real-time PCR assay. Using cultured cell lines, the real-time TRAP showed a dramatic improvement in the reliability and accuracy of quantitation of telomerase activity and was able to discriminate the A549 cells from hundreds-fold human embryonic lung cells. Using clinical samples of 60 lung cancers and 8 inflammatory lesions, the real-time TRAP was also superior in quantitation, high-throughput capability and standardization. Our modified real-time TRAP should be applicable for the detection of telomerase activity for the initial screening and progression monitoring of lung cancer patients. Our approach is particularly useful when only limited clinical specimen is available, such as fine needle aspiration or other cytological specimens that may contain only a small number of tumor cells.

[Expression of angiotensin II receptor subtypes in atrial fibrillation underlying rheumatic heart disease].

OBJECTIVE: To examine the expression of angiotensin II (Ang II) receptor subtypes in human left and right atrial tissue in atrial fibrillation underlying rheumatic heart disease. METHODS: Atrial tissue samples were obtained from 39 patients with rheumatic heart disease, 25 with atrial fibrillation (AF) and 14 with sinus rhythm(SR) during open heart surgery. AT1 and AT2 mRNA levels were measured with semi-quantitative reverse transcription polymerase chain reaction techniques. AT1 and AT2 protein levels were measured with immunohistochemical techniques. RESULTS: Compared with that of the SR group, left atrial inner diameter was significantly increased in the patients of the AF group. The AT1 mRNA and protein levels in the LA significantly increased in patients with AF compared with those in patients with SR (P < 0.05), whereas AT2 mRNA and protein were not significantly altered. Investigations of Ang II receptor subtypes' mRNA and protein levels in the RA did not exhibit any significant changes either in AT1 or AT2 in patients with AF and SR. CONCLUSIONS: AF is associated with an up-regulation of AT1 in LA, but does not appear to influence the AT2 expression. This may indicate a possible pathophysiologic role for renin-angiotensin system in the development of AF. The series of effects mediated by AT1 activation may be one of the molecular mechanisms involved in the process of atrial remodeling.

EGFR mutations are more frequent in well-differentiated than in poor-differentiated lung adenocarcinomas.

Somatic mutations in epidermal growth factor receptor (EGFR) tyrosine kinase domain, particularly deletions in exon 19 and point mutation in exon 21, are associated with clinical outcome in patients with lung adenocarcinoma, suggesting that EGFR mutation would have an important role in clinical decision making. DNA was extracted from the excised specimens of 60 lung adenocarcinoma patients with phenol-chloroform and ethanol precipitation. Exon 19 and 21 were amplified by PCR, and direct sequenced from both sense and antisense directions. EGFR somatic mutations were present in 13 of 60 patients (21.67%), including seven cases of in-frame deletion in exon 19 around codon 746 and six cases of amino acid substitution in exon 21. Exon 21 mutation is more frequent in adenocarcinomas with bronchi-alveolar component than exon 19 deletions. Mutations were more prevalent in well-differentiated adenocarcinomas (9/27, 33.33%) than in moderate to poor-differentiated adenocarcinomas (4/33, 12.12%) (P < 0.05). Adenocarcinomas with bronchi-alveolar components had higher mutation frequency (8/22,36. 36%) than those without bronchi-alveolar components (5/38, 13.16%) (P < 0.05). In this study, female patients had more mutation rate than male patients. This trend was also observed in the patients with pathologic stage I-II compared with stage III-IV, but neither of them was statistically significant. Patients with cisplatin-based adjuvant chemotherapy had no significantly prolonged survival compared with single radical resection. But patients with EGFR mutation had relative longer survival. In conclusion, our study suggest that EGFR mutations may be a valuable prognostic factor for disease free survival of surgically treated lung adenocarcinoma patients independently from adjuvant chemotherapy.

[Status and clinicopathologic implication of epidermal growth factor receptor mutation in non-small cell carcinoma of lung].

OBJECTIVE: To investigate mutations of epidermal growth factor receptor (EGFR) exon 19 and 21 in non-small cell lung carcinoma and to explore their clinicopathological correlations. METHOD: DNA was extracted from the excised tumor specimens of 66 non-small cell lung carcinoma patients by traditional phenol-chloroform and ethanol precipitation. Exons 19 and 21 were amplified by polymerase chain reaction (PCR), followed by direct sequencing in both sense and antisense directions. RESULTS: EGFR somatic mutations were present in 11 of 66 patients (16.7%), including 7 cases of in-frame deletion involving exon 19 and 4 cases of amino acid substitution involving exon 21. Mutations were more frequently observed in women (9/34, 26.5%) than in men (2/32, 6.3%), in adenocarcinomas (10/43, 23.3%) than squamous (0/13) and adenosquamous carcinomas (1/10). There was no difference in the mutation rates between smokers and non-smokers. Those with adenocarcinoma with bronchiolo-alveolar carcinoma (BAC) components had higher frequency of EGFR mutation (6/11) than those without non-BAC element (4/32, 12.5%). CONCLUSIONS: The mutations appear to occur in highly selected subgroups of lung cancer patients: adenocarcinomas with BAC components and patients of the female gender. The results may offer practical approach to the rapid identification of lung cancer patients who likely respond to EGFR inhibitor therapy.

[Detection of Mycobacterium tuberculosis DNA in sarcoidosis samples using real-time fluorescence polymerase chain reaction].

OBJECTIVE: To study the role of Mycobacterium tuberculosis in the pathogenesis of sarcoidosis. METHODS: Archival material of 22 patients with a histologic diagnosis of sarcoidosis were retrieved. Real-time fluorescent polymerase chain reaction (PCR) was used to detect DNA fragments of the complex-specific insertion sequence IS6110 of Mycobacterium tuberculosis in formalin-fixed and paraffin-embedded biopsy samples. RESULTS: Among the 22 samples studied, Mycobacterium tuberculosis DNA was detected in 11 cases. The sequence of PCR amplified IS6110 DNA fragments completely matched with the related sequence in Mycobacterium tuberculosis gene. CONCLUSIONS: Mycobacterium tuberculosis DNA is identified in a certain proportion of patients with a clinicopathologic diagnosis of sarcoidosis. Mycobacterium tuberculosis may be an important etiologic agent, at least in some of these patients.

[Diagnosis of lymph node micrometastases in human lung carcinoma: determination of p53 gene mutations by polymerase chain reaction-temperature gradient gel electrophoresis (PCR-TGGE)].

BACKGROUND & OBJECTIVE: Metastasis is one of the most important factors in determining the prognosis of lung cancer patients. However, it is difficult to determine single cancer cell in lymph nodes by routine methods. This study was designed to investigate a sensitive method for detecting lymph node micrometastases in human lung carcinomas. METHODS: Mutation detection for p53 gene (exon 5-8) was performed in 39 cases of primary lung carcinomas and 110 corresponding lymph nodes (LNS) by PCR-TGGE method. Serial sectioning was performed to confirm micrometastases in some of these samples. RESULTS: p53 mutations were found in 23 of 39 cases with primary lung cancer. Forty LNSs showed p53 mutation in 67 corresponding LNSs. They were located at the same exon as their primary tumors. Only twenty-six LNSs were detected metastases by routine histopathology. Forty-three LNSs from sixteen cases of primary tumor without p53 mutation and ten LNSs from the patients without tumor disease did not have any p53 mutations. By PCR-TGGE assay p53 gene mutations were determined in fourteen LNSs without histopathological evidence of metastasis. Micrometastases were found in four of the serially sectioned cases. CONCLUSION: Mutation detection for p53 gene (exon 5-8) showed lymph node micrometastases in patients with lung carcinoma. It may be used as a complement for the routine histopathology.