Objective Microheterogeneity is an important problem in molecularly biological and proteomic research on malignancies.The goal of this study was to investigate the method to purify targeted cells in lung cancer and its paired normal tissues for proteomic study on lung cancer applying laser capture microdissection(LCM).Methods LCM technique was emploged to obtain the cells of lung cancer tissues and their paired normal tissues from frozen sections.Results LCM can be applied to quickly and precisely obtain pure targeted cells subgroup or single cell without interstitium under the microscope.Conclusion LCM can tackle the problem of tissue heterogeneity in molecular analysis successfully.These results indicate that LCM has a potential as a tool in lung cancer proteomic research.

BACKGROUND: It has been widely reported that the most types of cancer are probably originated from cancer stem cells (CSCs) which are subpopulations of tumor cells. Recent studies also suggested that lung cancer could arise from CSCs. However, the phenotypic characteristics of CSCs in lung cancer have not been precisely described.OBJECTIVE: To systematically analyze the expression of the most common CSC markers in cell line A549.METHODS: A549 cells were cultured for 1 week under the condition of conventional high glucose. After that, flow cytometry was used to assess the expression of putative stem cell markers, including CD133, CD24, CD44 and ABCG2.Cells were then sorted according to the expression of CD44 and CD24 markers by fluorescence-activated cell sorting (FACS) and characterized using their sphere-forming capacity in serum free medium supplemented with several growth factors.RESULTS AND CONCLUSION: A549 cells expressed the CSC markers CD44 and CD24 at 64.23％ and 58.62％,whereas the expression of both ABCG2 and CD133 was around 0.9％. Double-positive CD44/133 populations were rare.CD44+/CD24+ and CD44+/CD24- subpopulations respectively exhibited 54.64％ and 23.38％ expression. CD44+/CD24+ and CD44+/CD24- subsets were sorted by FACS. Both isolated subpopulations formed spheres in the serum free medium supplemented with basic fibroblast growth factor and epidermal growth factor. However, there was no significant difference in the sphere formation efficiency among CD44+/CD24+ and CD44+/CD24- subsets as well as A549 cells. Our findings suggest that CD44 and CD24 cannot be considered as potential markers for isolating lung CSCs in cell line A549, and further investigation using in vivo assays is required.

Objective To investigate the influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells.Methods The S100A6 gene RNA interference vector was transfected in A549 lung adenocarcinoma by lentivirus.The experiment was divided into three groups:pLenR-GPH group (the vector without S100A6 RNAi gene was transfected),negative control group (no vectors was transfected),and RNAi group (the vector with S100A6 RNAi gene was transfected).S100A6 mRNA and protein were detected using real-time PCR and Western blotting.The biological behavior including cell proliferation,invasion,cell cycle and cell apoptosis were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide,transwell,and flow cytometer,respectively.Results The expression of S100A6 mRNA of A549 lung adenocarcinoma cell line in RNAi group (0.009 ± 0.001) was significantly decreased than those in negative control group (0.049 ± 0.005) and pLenR-GPH group (0.030 ± 0.006),with statistically significant differences (t =57.56,P =0.000;t =48.21,P =0.000).The expression of S100A6 protein of A549 lung adenocarcinoma cell line in RNAi group (0.107 ± 0.002) was significantly decreased than those in negative control group (0.341 ± 0.005) and pLenR-GPH group (0.311 ± 0.006),with statistically significant differences (t =37.34,P =0.000;t =27.51,P =0.001).The ability of cell proliferation at 48 hours in RNAi group (0.230 ± 0.008) was significantly declined than those in negative control group (0.292 ± 0.038) and pLenR-GPH group (0.307 ± 0.013),with statistically significant differences (t =25.31,P =0.003;t =29.42,P =0.001).The number of transmembrane cells in RNAi group (11.40 ± 1.36) was significantly declined than those in negative control group (26.80 ± 1.83) and pLenR-GPH group (25.80 ± 1.93),with statistically significant differences (t =29.44,P =0.001;t =23.17,P =0.005).The cell proportion of S phase in RNAi group (28.26％ ± 0.38％) was significantly lower than those in pLenR-GPH group (44.73％±0.66％) and negative control group (45.15％ ± 1.69％),with statistically significant (t =63.69,P=0.000;t =71.55,P =0.000).Cell propotion of G2-M phase in RNAi group (26.99％ ± 0.29％) was signi-ficantly higher than those in negative control group (13.26％ ±0.49％) and pLenR-GPH group (12.41％ ± 0.46％),with statistically significant (t =56.31,P =0.000;t =51.39,P =0.000).The cell apoptosis proportion in RNAi group (8.90％ ±0.48％) was significantly higher than those in negative control group (5.84％ ±0.21％) and pLenR-GPH group (5.99％ ±0.37％),with statistically significant (t=51.34,P =0.000;t =47.27,P =0.000).Conclusion S100A6 gene involves the proliferation,invasion,cell cycle and apoptosis of tumor cells,which has close correlation with occurrence,development and metastasis of lung adenocarcinoma.S100A6 gene is hopeful to become a molecular target for the diagnosis and treatment of lung adenocarcinoma.

10?g/L may be very valuable for diagnosis of lung cancer, especially for adenocarcinoma. Cyfra21-1 and NSE were better biomarkers for squamous cell carcinoma and SCLC, respectively. The combined detection of Cyfra21-1 and CEA could be used as a better pattern for diagnosis of lung cancer.

BACKGROUNDTo study the expression and clinicopathological significance of CD44V6 in non-small cell lung cancer (NSCLC).

METHODSCD44V6 expression in lung cancer tissues was detected in 90 patients with lung cancer by immunohistohemistry SABC method.

RESULTSThe positive rate of CD44V6 expression in NSCLC was 51.1% (46/90), which was related to TNM stage, but not to cell differention, age and sex of the patients (P > 0.05). The positive rate of CD44V6 expression was 32.2% (10/31) in stage I+II patients and 61.0% (36/59) in stage III+IV disease. A highly significant difference was existed between the two groups (P < 0.01). The positive rate of CD44V6 expression was significantly higher in NSCLC with lymph node metastasis (61.0%, 36/59) than that without lymph node metastasis (32.2%, 10/31) (P < 0.01). The 3-year survival rate (65.5%, 19/29) in patients with low CD44V6 expression was significantly higher than that (35.0%, 7/20) with high CD44V6 expression (P < 0.05).

CONCLUSIONSOverexpression of CD44V6 may be involved in the progression and metastasis of the cancer and prognosis of the patients with lung cancer. It might be helpful to evaluate the progression of the cancer, and to predict the metastasis and progression of lung cancer.

Biopsy, Fine-Needle ; Bronchi ; diagnostic imaging ; pathology ; Endosonography ; adverse effects ; methods ; Humans ; Thoracic Neoplasms ; diagnostic imaging

Since mid-December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has outbroken in Wuhan, Hubei Province, China, and spread rapidly to other provinces in China and dozens of countries and regions around the world, becoming the Public Health Emergency of International Concern (Public Health Emergency of International Concern). SARS-CoV-2 can mainly transmit by droplets or close contact, and is generally susceptible in the crowd. Tumor patients are at high risk of this pathogen because of their impaired immune function. Identifying tumor patients with 2019 novel coronavirus disease (COVID-19) early, and understanding its distribution characteristics can help to improve the cure rate of patients, and better control the epidemic and development of SARS-CoV-2 much better. With comprehensive analysis of relevant literature, this paper reviews the clinical characteristics of neoplastic patients with COVID-19, and puts forward some suggestions on how to deal with this epidemic.
Betacoronavirus ; Coronavirus Infections ; complications ; epidemiology ; prevention & control ; transmission ; Epidemics ; Humans ; Neoplasms ; complications ; Pandemics ; prevention & control ; Pneumonia, Viral ; complications ; epidemiology ; prevention & control ; transmission

BACKGROUND: A lack of effective treatment for lung squamous cell carcinoma (LUSC) makes it an important factor restricting the 5-year survival rate of non-small cell lung cancer (NSCLC). Long non-coding RNA 00668 (LINC00668) was reported to play crucial regulatory roles in the tumorigenesis and progression of various cancers; however, its role in LUSC is unclear. The aim of this study was to investigate the prognosis value and biological function of LINC00668 in NSCLC, especially in LUSC. METHODS: The expression pattern of LINC00668 and its relationship with clinical characteristics and prognosis of patients were investigated in the NSCLC especially LUSC based on The Cancer Genome Altas (TCGA) database. Its function in LUSC cells was explored in vitro. RESULTS: LINC00668 expression was significantly up-regulated in LUSC patients and high expression level of LINC00668 was associated with advanced tumor-node-metastasis (TMN) stage. Moreover, the expression of LINC00668 significantly increased in smoking patients, and was a prognostic indicator for overall survival (OS) of smoking patients with LUSC. In vitro experiments showed that LINC00668 has significantly higher expression level in LUSC cell lines and tissues compared to normal bronchial epithelial cell and para-tumor tissues; meanwhile, functional assay indicated knockdown of LINC00668 effectively inhibited the migration and invasion of LUSC cells. CONCLUSIONS LINC00668 might closely relate to the development of LUSC, and inhibition of LINC00668 may reduce the metastasis of LUSC.
Carcinoma, Non-Small-Cell Lung ; Carcinoma, Squamous Cell/genetics* ; Cell Movement/genetics* ; Humans ; Lung ; Lung Neoplasms/genetics* ; RNA, Long Noncoding/genetics*

BACKGROUND: Solitary pulmonary nodule has received increasing attention in recent years. A couple of lung nodules have been recognized as primary malignant tumors, which leads to an urgent need in enhancing the diagnosis of benign/malignant lung nodules at clinical settings. This study aims to explore the value of the combined detection of cytokines and tumor markers in differencing benign and malignant solitary pulmonary nodules in diagnose. METHODS: With 81 solitary pulmonary nodules cases with a clear diagnosis, the general clinical data, nodule imaging features, pathological diagnosis data, serological index cytokine series and tumor marker expression levels were collected in groups. Both single factor and multi-factors analysis were conducted to screen out the serum influence indexes that can predict the malignant probability of lung nodules, and mean while binary logistic regression analysis was used to construct joint indexes; After receiver operating characteristic curve (ROC) was drawn, the area under the curve and the corresponding sensitivity, specificity and positive of each index predicted value, negative predicted value and accuracy could be calculated with a view to determine the statistical significance of area under the curve (AUC). RESULTS: There are differences in the distribution of malignant solitary pulmonary nodules at different locations, with the highest proportion of the right upper lobe (40.4%). The serum levels of carcinoembryonic antigen (CEA), cytokeratin 19 fragment 21-1 (CYFRA21-1), interleukin-6 (IL-6), interleukin-8 (IL-8) in the malignant nodule group were higher than those in the benign nodule group. Logistic regression analysis suggests that CEA, IL-6 and IL-8 are independent risk factors for predicting malignant nodules. ROC curve analysis shows that the areas under the curve of the individual indicators CEA, IL-6 and IL-8 are 0.642, 0.684 and 0.749. The comparison result of the test efficiency of the area under the curve suggests that CEA+IL-6+IL-8 has a larger area under the curve and higher detection efficiency. CONCLUSIONS CEA, IL-6 and IL-8 are independent risk factors for malignant solitary pulmonary nodules. The combined detection of cytokines and tumor markers has played a role in the differential diagnosis of benign and malignant lung nodules. The diagnostic value of the combined detection of CEA+IL-6+IL-8 is the highest.