OBJECTIVETo investigate DNA-dependent protein kinase catalytic subunit (DNA-PKcs) content and activity in lung adenocarcinoma cell lines and its correlation with radiosensitivity.

METHODSThe content and activity of DNA-PKcs were analyzed in two lung adenocarcinoma cell lines A549 and H1299 by Western blotting and the Signa TECT DNA-PK assay kit. The dose-survival relationship for two cell lines was analyzed using clonogenic formation assay.

RESULTSA549 was more radiosensitive than H1299. The survival fractions at 2 Gy (SF2) were 0.7412 in A549 cell line and 0.2473 in H1299 cell line. The content of DNA-PKcs was significantly higher in A549 cells than in H1299 cells (t=10.37, P<0.001). The integrated optical densities were 3.29-/+0.44 in A549 cells and 0.50-/+0.17 in H1299 cells. DNA-PKcs activities in A549 and H1299 cells were 8.29-/+1.37 and 2.47-/+1.09, respectively, showing a significant difference between them (t=5.76, P=0.005).

CONCLUSIONDNA-PKcs is an important factor to affect the radiosensitivity of lung adenocarcinoma cell lines.

Adenocarcinoma ; enzymology ; pathology ; Calcium-Binding Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; enzymology ; pathology ; Radiation Tolerance

OBJECTIVETo evaluate the application of traditional cytomorphology, telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis of pleural effusion and bronchoalveolar lavage samples.

METHODSA 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.

RESULTSTelomerase 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.

CONCLUSIONSTelomerase 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.

Breast Neoplasms ; diagnosis ; enzymology ; pathology ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Lung Neoplasms ; diagnosis ; enzymology ; pathology ; Pleural Effusion ; diagnosis ; enzymology ; pathology ; Pleural Effusion, Malignant ; diagnosis ; enzymology ; pathology ; Sensitivity and Specificity ; Telomerase ; metabolism

OBJECTIVETo investigate the expression of tripartite-motif protein 25 (TRIM25) and pyruvate kinase M2 (PKM2) protein in non-small cell lung cancer (NSCLC) and explore their role in the occurrence and progression of NSCLC.

METHODSThe expressions of TRIM25 and PKM2 protein were detected in 60 NSCLC specimens and 20 adjacent normal lung tissue (>5 cm from the lesions) with immunofluorescence histochemical method and in 10 fresh specimens of NSCLC with Western blotting. The results were analyzed in relation with the clinicopathological features of the patients.

RESULTSThe positivity rates of TRIM25 expression was 45% in the 60 lung carcinoma specimens, significantly higher than that in the 20 normal lung tissues (10%, P=0.005). TRIM25 protein was expressed in 28.6% of lung adenocarcinoma tissues and in 59.4% of squamous carcinoma tissues (P=0.017). TRIM25 protein expression was positively correlated with the TNM stages and lymph node metastasis of NSCLC (P<0.05). The expressions of PKM2 protein in 60 cases of lung carcinoma was 73.3%,while in 20 cases of normal lung tissues the expressions was 30%(P=0.001). The positivity rates of PKM2 expression differed significantly between lung adenocarcinoma and squamous carcinoma (57.1% vs 87.5%, P=0.008). An inverse correlation was noted between TRIM25 and PKM2 expressions (P=0.026).

CONCLUSIONTRIM25 and PKM2 protein may participate in the occurrence and progression of NSCLC, and their expressions are inversely correlated.

Adenocarcinoma ; enzymology ; Carcinoma, Non-Small-Cell Lung ; enzymology ; Carcinoma, Squamous Cell ; enzymology ; Carrier Proteins ; metabolism ; Humans ; Lung ; pathology ; Lung Neoplasms ; enzymology ; Lymphatic Metastasis ; Membrane Proteins ; metabolism ; Thyroid Hormones ; metabolism ; Transcription Factors ; metabolism ; Tripartite Motif Proteins ; Ubiquitin-Protein Ligases ; metabolism

OBJECTIVETo investigate the effect of DNA methyltransferases 1 (DNMT-1) inhibition on the ACC-M cells in vitro and in vivo and discuss the role of DNMT-1 in the development, invasion and metastasis of salivary adenoid cystic carcinoma (SACC).

METHODSACC-M cells of stable DNMT-1 inhibition were established in a previous research. In vitro, the growth and invasion of ACC-M cells which stably inhibited DNMT-1 were detected and analyzed by methyl thiazolyl tetrazolium (MTT) growth curve, flow cytometry, plating efficiency and invasion assay. In vivo, the growth and metastasis of ACC-M cells which persistently inhibited DNMT-1 were observed and analyzed by subcutaneous injection and tail vein injection into the nude mice.

RESULTSIn vitro, the doubling time [(34.7 +/- 2.1) h], S phase fraction [(17.4 +/- 1.7)%], plating efficiency [(43.0 +/- 1.3)%] of ACC-M cells was significantly different from those of blank [(26.2 +/- 3.1) h, (31.5 +/- 2.0)%, (71.0 +/- 4.7)%], empty load control [(28.4 +/- 3.9) h, (39.0 +/- 2.0)%, (66.0 +/- 5.2)%], P < 0.05, and the invasion ability was not significantly different among these groups (P > 0.05). In vivo, the subcutaneous tumor forming rate (6/10), volume [(2.18 +/- 0.83) mm(3)], weight [(0.0156 +/- 0.0046) g] of ACC-M cells was also significantly lower than that of blank [10/10, (155.44 +/- 1.67) mm(3), (0.0724 +/- 0.0157) g], empty load control [10/10, (147.46 +/- 1.73) mm(3), (0.0729 +/- 0.0177) g], P < 0.05, but the rate of lung metastasis was not significantly different among these groups (P > 0.05), and the masses (2.0 +/- 0.5), diameter (70.0 +/- 20.3) microm of ACC-M cells was significantly lower than that of blank [(28.0 +/- 5.5), (195 +/- 25.4) microm], empty load control [(27.0 +/- 4.5), (190.0 +/- 19.9) microm], P < 0.05.

CONCLUSIONSInhibition of DNMT-1 is able to inhibit the proliferation and metastasis of ACC-M cells in vitro and in vivo.

Animals ; Carcinoma, Adenoid Cystic ; enzymology ; pathology ; secondary ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; secondary ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Repressor Proteins ; antagonists & inhibitors ; Salivary Gland Neoplasms ; enzymology ; pathology

OBJECTIVETo study the effects of neuron specific enolase (NSE) gene silencing on the cell proliferation and apoptosis of lung cancer cells in vitro.

METHODSNSE protein expression was detected in human small cell lung cancer cell line NCI-H446 and non-small cell lung cancer cell line A549 by immunocytochemistry, and a small interference RNA (siRNA) was transfected into the cells to inhibit NSE gene expression. The changes in the cell cycle, apoptosis, Ki67 protein and caspase-3 activity in the transfected cells were observed by flow cytometry, Western blotting and colorimetric assay, respectively.

RESULTSBoth A549 and NCI-H446 cells expressed NSE protein. Transfection of siRNA for NSE gene significantly inhibited the expression of NSE gene in the cells, resulting in an inhibition rate exceeding 90%. NSE gene silencing caused significantly decreased cell percentage in S phase and the expression of Ki67 protein, and increased the cell apoptotic rate and caspase-3 activity.

CONCLUSIONNSE gene expression promotes the cell proliferation and inhibits the cell apoptosis in lung cancer cells with neuroendocrine differentiation, which might be a causative factor contributing to increased malignancy of the cells.

Adenocarcinoma ; enzymology ; genetics ; pathology ; Apoptosis ; genetics ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Ki-67 Antigen ; metabolism ; Lung Neoplasms ; enzymology ; genetics ; pathology ; Phosphopyruvate Hydratase ; genetics ; RNA Interference ; Small Cell Lung Carcinoma ; enzymology ; genetics ; pathology

OBJECTIVETo determine the cut-off value of serum neuron-specific enolase (NSE) level for distinguishing small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC).

METHODSSerum NSE levels were measured by enzyme-linked immunosorbent assay in 137 patients with NSCLC or SCLC, and the best cut-off value was analyzed using ROC curve.

RESULTSThe positivity rate of serum NSE was significantly higher in patients with SCLC than in those with NSCLC (P<0.01). The best cut-off value was 15.45 microg/L using ROC curve, which gave a sensitivity of 66.7% and specificity of 65.7%.

CONCLUSIONSerum NSE level may allow simple and cost-effective differentiation of SCLC and NSCLC.

Biomarkers, Tumor ; blood ; Carcinoma, Non-Small-Cell Lung ; enzymology ; pathology ; Carcinoma, Small Cell ; enzymology ; pathology ; Diagnosis, Differential ; Female ; Humans ; Lung Neoplasms ; enzymology ; pathology ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; blood

OBJECTIVETo investigate the expression and its significance of DNA topoisomerase I (Topo I) in small cell lung cancer (SCLC).

METHODSTopo I expression was detected by immunohistochemical S-P technique on 50 cases of SCLC and 12 cases of normal lung tissues.

RESULTSThe total positive rate of Topo I in normal lung tissue was 25.0% (3/12) and 78.0% (39/50) in SCLC. The expression of Topo I does not correlate with age, gender, smoking, tumor size and tumor site (P > 0.05), but significantly correlated with lymph node metastasis and clinical stage (P <0.05).

CONCLUSIONHigh Topo I expression is a rationale indication of Topo I inhibitor treatment of malignancies. It should be possible to predict anti-tumor drug sensitivity by assessment of Topo I expression and help to improve the therapeutic efficacy for cancer patients.

Carcinoma, Small Cell ; enzymology ; pathology ; DNA Topoisomerases, Type I ; metabolism ; Female ; Humans ; Immunohistochemistry ; methods ; Lung Neoplasms ; enzymology ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVETo assess protein and mRNA expression levels of heparanase and basic fibroblast growth factor (bFGF) genes in human non-small cell lung cancer (NSCLC) and their roles in tumor invasion, metastasis and prognosis.

METHODSA total of 115 paraffin-embedded and 45 fresh-frozen tissue specimens of NSCLC were studied by immunohistochemistry, Western Blot and in situ hybridization to evaluate the protein and mRNA expression status of heparanase and bFGF genes. The data was analyzed by SPSS statistical software.

RESULTSBoth human heparanase and bFGF were highly expressed in NSCLC cells, in contrast to none or a low expression in normal lung tissue. Expression of heparanase also showed a significantly higher than that in the normal tissue by Western blot (P = 0.041). Immunohistochemistry showed that heparanase expression was both cytoplasmic and membranous. The agreement between heparanase and bFGF was significant. A significant correlation was found between the expression of either protein and TNM stage, vascular invasion, lymphatic metastasis and microvascular density (MVD). Co-expression of the two proteins demonstrated an even higher correlation with the tumor stage and MVD. In addition, expression of bFGF correlated with tumor cell differentiation. Data of a multivariate analysis indicated that tumor cell differentiation, vascular invasion, lymphatic metastasis and expression of bFGF were identified as significant prognostic parameters.

CONCLUSIONSBoth heparanase and bFGF may play important roles in tumor angiogenesis, metastasis, and prognosis of NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; enzymology ; metabolism ; pathology ; Cell Differentiation ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Glucuronidase ; metabolism ; Humans ; Lung Neoplasms ; enzymology ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; Prognosis ; Survival Rate

OBJECTIVETo study the relations among COX-2 expression in non-small cell lung cancer (NSCLC), its clinical characteristics and brognosis.

METHODSImmunostaining was performed with COX-2 antibody to the surgically resected tissue samples from 79 patients with NSCLC. Vessel epithelium cell COX-2 expression was taken as positive control.

RESULTSThe positive rate of adenocarcinoma and squamous cell carcinoma was 85% and 57%, respectively (P = 0.013). COX-2 expression was associated with the extent of adenocarcinoma differentiation, tumor size, and TNM period, but not with the extent of squamous cell carcinoma differentiation. In the COX-2 positive group, the 5-year survival rate and a median survival time were 27.1% and 53 months; however in the negative group they were 52.0% and 61 months (P = 0.029).

CONCLUSIONInvasive development of NSCLC is related to inceased expression of COX-2. COX-2 overexpression may be one of the risky factors for the prognosis of NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; enzymology ; mortality ; pathology ; Cyclooxygenase 2 ; Female ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Lung Neoplasms ; enzymology ; mortality ; pathology ; Male ; Membrane Proteins ; Middle Aged ; Prognosis ; Prostaglandin-Endoperoxide Synthases ; analysis ; Survival Rate

BACKGROUND: Epidermal growth factor receptor (EGFR)-based targeted therapy improves the survival of patients with advanced lung adenocarcinoma harboring EGFR mutations. However, factors including treatment or heterogeneity partly contribute to EGFR genetic status alteration between baseline and disease progresses (PD). The aim of this study is to compare difference of EGFR mutations between biopsy and rebiopsy in real world. METHODS: Data from 61 paired specimens performed EGFR testing in Jilin Provincial Cancer Hospital between January 2015 and December 2017 were collected and analyzed. The specimens were collected at baseline and PD, confirmed by histology or cytology and categorized as tumor tissue, malignant pleural effusion or plasma. All patients were naive and received chemotherapy or targeted therapy as first-line treatment. Amplification Refractory Mutation System (ARMS) was used to detect EGFR mutations. RESULTS: EGFR mutation rate in tumor tissue, pleural effusion or blood was 90.2% vs 88.5%, 6.6% vs 6.6% and 3.2% vs 4.9% at baseline or PD respectively and discrepancy was 72% and 36.3% for the same (n=50) or different (n=11) type of specimens. The EGFR mutation rate was 95.1% and 91.8% in patients before and after treatment, and the discrepancy was 63.9%, among which, 69.2% and 92.3% in chemotherapy-treated patients (n=13) with discrepancy to 46.1% (6/13), and 100.0% and 91.7% in EGFR-TKI-treated patients (n=48) with discrepancy to 70.8%. There were four types of alterations in terms of EGFR mutations: wild type turned into mutation (4.9%), mutation disappeared (8.2%), sensitive mutations transformed (1.6%), and new mutations appeared (49.1%). CONCLUSIONS In real world, the EGFR mutation status in advanced non-small cell lung cancer (NSCLC) patients altered significantly, due to tissue resources and therapeutic approaches, implying the importance of rebiopsy and real-time detection of EGFR mutation, in order to provide data to guide precise strategy in the following treatment.
Adult ; Aged ; Biopsy ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; enzymology ; genetics ; pathology ; ErbB Receptors ; antagonists & inhibitors ; genetics ; Female ; Humans ; Lung Neoplasms ; drug therapy ; enzymology ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Protein Kinase Inhibitors ; therapeutic use ; Retrospective Studies ; Treatment Outcome