Lung cancer is the leading cause of cancer-related deaths worldwide. Non-small cell lung cancer (NSCLC) accounts for 80%-85% of all patients with lung cancer, the majority of patients with lung cancer at the time of diagnosis is in the advanced stage. The development of target therapy based on has changed the mode of treatment in patients with advanced NSCLC. In NSCLC, epidermal growth factor receptor mutation (EGFR) fusion with echinoderm microtubule-associated protein-like4-anaplastic lymphoma kinase (EML4-ALK) has been shown to be a powerful biomarker. It is well known that KRAS is also NSCLC one of the most common mutations in oncogenes, although more than 20 years ago KRAS mutation was found in NSCLC. At present, although there are many drugs used to treat NSCLC patients with KRAS mutation, there is no selective or specific inhibitor for the direct elimination of KRAS activity. NSCLC patients with KRAS mutation have poor responsiveness to most systemic therapy. However, individualized therapy for activated signaling pathways with targeted drugs has a good effect on the prognosis of NSCLC patients with KRAS mutation. In addition, the prognostic and predictive role of KRAS mutation in NSCLC remains unclear. In this review, we focus on the research progress of NSCLC with KRAS mutation, including molecular biology, clinicopathological features, prognosis and prediction of KRAS mutation, which will help to improve the understanding of NSCLC in KRAS mutation.
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Animals ; Carcinoma, Non-Small-Cell Lung ; enzymology ; genetics ; Humans ; Lung Neoplasms ; enzymology ; genetics ; Mutation ; Proto-Oncogene Proteins p21(ras) ; genetics

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 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 investigate the changes in serum protease and cytokine in patients with silicosis, tuberculosis, and lung cancer.

METHODSSerum samples of patients with silicosis, tuberculosis, and lung cancer were collected. The variation trends of the expression of granzyme A, cathepsin G, apolipoprotein A, and interferon-β (IFN-β) were analyzed using enzyme-linked immunosorbent assay.

RESULTSThe concentration of apolipoprotein A of the silicosis group was 200 µg/ml, significantly higher than those of the tuberculosis and lung cancer groups (P < 0.05), and the lung cancer group had a significantly higher concentration of apolipoprotein A compared with the tuberculosis group (P < 0.05). The silicosis group had significantly higher expression of cathepsin G compared with the tuberculosis and lung cancer groups (P < 0.05), and the tuberculosis group and lung cancer group showed no significant difference in the concentration of cathepsin G (P > 0.05). The tuberculosis group had a significantly higher concentration of granzyme A than the silicosis and lung cancer groups (P < 0.05), and the silicosis group and lung cancer group had similar protein concentration trends (P > 0.05). The tuberculosis group and lung cancer group had significantly higher concentration of IFN-β compared with the silicosis group (P < 0.05), and the tuberculosis group and lung cancer group showed no significant difference in IFN-β concentration (P > 0.05).

CONCLUSIONThis study may offer diagnostic markers for the clinical diagnosis of silicosis, tuberculosis, and lung cancer, and could provide a basis for the research, as well as potential molecular targets for the diagnosis and treatment of these diseases.

Biomarkers ; Cathepsin G ; metabolism ; Cytokines ; blood ; Endopeptidases ; blood ; Enzyme-Linked Immunosorbent Assay ; Granzymes ; metabolism ; Humans ; Interferon-beta ; metabolism ; Lung Neoplasms ; enzymology ; Silicosis ; enzymology ; Tuberculosis ; enzymology

BACKGROUND AND OBJECTIVEIt has been proven that telomerase activation correlates with the carcinogenesis, aggressiveness and turnover of lung cancer. Telomerase is one of the improtant molecular biomarkers for diagnosis and targeting therapy in lung cancer. The aim of this study is to investigate the diagnostic value of the combined determination of telomerase activity in induced sputum, pleural effusion and fiberobronchoscopic biopsy in lung cancer patients.

METHODSThe technique of TRAP (telomeric repeat amplification protocal)-PCR-ELISA was employed to detect telomease levels of induced sputum, pleural effusion and fiberobronchoscopic biopsy in 80 lung cancer patients with pleural effusion and 50 benign pulmonary disease patients with pleural effusion.

RESULTSTelomemse levels of induced sputum, pleural effusion and fiberobronchoscopic biopsy were all significantly higher in patients with lung cancer than those with benign pulmonary disease (P < 0.001). There was no significant difference in the level of telomerase activity between different pathologic types (P > 0.05). The sensitivity of induced sputum, pleural effusion and fiberobronchoscopic biopsy were 62.5% (50/80), 46.3% (37/80) and 60.0% (48/80), respectively. The specificity were 72.0% (36/50), 66.0% (33/50) and 70.0% (35/50), respectively. The overall accuracy were 66.2% (86/130), 53.8% (70/130) and 63.8% (83/130), respectively. The sensitivity, specificity and overall accuracy of combined induced sputum, pleural effusion and fiberobronchoscopic biopsy were 85.0% (68/80), 78.0% (39/50) and 82.3% (107/130), respectively. The sensitivity of telomease level in combined detection for diagnosis of lung cancer was much higher than that in single sample detection (P < 0.01).

CONCLUSIONThe sensitivity of telomease activity in combined three samples was the highest. It can further improve the accuracy for the diagnosis of lung cancer with pleural effusion.

Aged ; Biopsy ; Bronchoscopy ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lung Neoplasms ; diagnosis ; enzymology ; Male ; Middle Aged ; Pleural Effusion ; enzymology ; Polymerase Chain Reaction ; Sputum ; enzymology ; Telomerase ; metabolism

OBJECTIVETo investigate the frequency of anaplastic lymphoma kinase (ALK) expression in non-small cell lung cancer (NSCLC) patients and its correlation with the clinicopathologic features.

METHODSALK immunohistochemistry and ALK fluorescent in situ hybridization (FISH) were performed on formalin-fixed, paraffin-embedded tissue in 100 cases of NSCLCs between 2011 and 2013. Relevant clinicopathologic data were collected and correlated with ALK expression.

RESULTSAll patients with immunohistochemical score of 3 (n = 12) were FISH-positive and all patients with score of 0 (n = 78) were FISH-negative. Among patients with immunohistochemical scores of 1 and 2, 2/3 and 6/7 were FISH-positive, respectively. The sensitivity and specificity of ALK immunohistochemistry with intensity score of 1 or more were 100% and 98%, respectively. Invasive mucinous adenocarcinoma, solid or acinar growth pattern, presence of mucous cells (signet-ring cells or goblet cells), extracellular mucus and lack of significant nuclear pleomorphism characterized ALK-rearranged cancer.

CONCLUSIONSALK-rearranged cancers possess specific histological features. Immunohistochemistry can be used as a routine test for screening ALK-positive cases in advanced NSCLC, and FISH testing should be used to confirm ALK translocation for patients with tumors showing staining for ALK by immunohistochemistry. All of these can help physicians identify patients who may benefit from targeted therapy.

Carcinoma, Non-Small-Cell Lung ; enzymology ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; enzymology ; Male ; Paraffin Embedding ; Receptor Protein-Tyrosine Kinases ; metabolism ; Sensitivity and Specificity

Molecular target therapy is one of the most popular field of non-small cell lung cancer (NSCLC) treatmnet. Epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) rearragement are the most important two oncogenic drivers in NSCLC, early studies suggested that EGFR mutations and ALK rearrangements are mutually exclusive, but isolated cases or small sample research with concomitant EGFR and ALK alterations have been constantly reported. The co-occurrence of EGFR mutations and anaplastic lymphoma kinase (ALK) rearrangements constitutes a rare molecular, the frequency of EGFR/ALK co-alterations was about 1%, however, little has been known about clinicopathologic feature and treatment. This review summarized published case report, EGFR and ALK alterations are common in female, Asian origin, never smoker, IV stage, and denocarcinomas. First-line treatment can choose EGFR or ALK tyrosine kinase inhibitors (TKIs). However, studies about the origin and resistance mechanism in EGFR/ALK co-alterations are little, require more experimental and clinical research.
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Anaplastic Lymphoma Kinase ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; enzymology ; genetics ; ErbB Receptors ; genetics ; Humans ; Lung Neoplasms ; diagnosis ; enzymology ; genetics ; Mutation ; Prognosis ; Receptor Protein-Tyrosine Kinases ; genetics

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 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