OBJECTIVETo study the over-expression of mutant p53 protein in non-mucinous adenocarcinoma in-situ (NMAIS) and invasive adenocarcinoma, NOS of lung.

METHODSImmunohistochemical study for p53 protein was performed on 17 cases of NMAIS and 70 cases of invasive adenocarcinoma, NOS of lung. The difference in p53 over-expression between the two tumor subtypes was analyzed.

RESULTSThe over-expression of mutant p53 protein was observed in 0 case (0%) of NMAIS and 37 cases (52.9%) of invasive adenocarcinoma, NOS of lung. The difference was of statistical significance (P = 0.000).

CONCLUSIONMutant p53 protein over-expression may play a role in the progression of NMAIS to invasive adenocarcinoma, NOS.

Adenocarcinoma ; metabolism ; Adenocarcinoma in Situ ; metabolism ; Humans ; Immunohistochemistry ; Mutant Proteins ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVETo identify differentially expressed microRNAs (miRNAs) related to lung adenocarcinoma in Xuanwei region and predict their target genes and related signaling pathways based on bioinformatic analysis.

METHODSHigh-throughput microarray assay was performed to detect miRNA expression profiles in 34 paired human lung adenocarcinoma and adjacent normal tissues (including 24 cases in Xuanwei region and 10 in other regions). Gene ontology and KEGG pathway analyses were used to predict the target genes and the regulatory signaling pathways.

RESULTSThirty-four miRNAs were differentially expressed in lung adenocarcinoma tissues in cases in Xuanwei region as compared with cases in other regions, including 23 upregulated and 11 downregulated miRNAs. The predicted target genes included GF, RTK, SOS, IRS1, BCAP, CYTOKINSR, ECM, ITGB, FAK and Gbeta;Y involving the PI3K/Alt, WNT and MAPK pathways.

CONCLUSIONThe specific microRNA expression profiles of lung adenocarcinoma in cases found in Xuanwei region allow for a better understanding of the pathogenesis of lung adenocarcinoma in Xuanwei. The predicted target genes may involve the PI3K/Alt, WNT and MAPK pathways.

Adenocarcinoma ; genetics ; metabolism ; Computational Biology ; Gene Expression Profiling ; Humans ; Lung ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; MicroRNAs ; genetics ; metabolism ; Signal Transduction

OBJECTIVETo investigate PTEN expression and mutation status in the development of cervical adenocarcinoma.

METHODSImmunohistochemistry study of PTEN protein was performed on 42 cases of cervical adenocarcinoma, 20 cases of cervical glandular intraepithelial neoplasia and 28 cases of normal cervix tissue samples. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to detect the presence of mutation of exons 5 and 8 of PTEN gene.

RESULTSPositive expression rates of PTEN protein were 54.8% (23/42), 25.0% (5/20) and 100% (28/28) in cervical adenocarcinoma, cervical glandular intraepithelial neoplasia and normal cervix tissues, respectively. There were significant differences among the 3 groups (P < 0.05). Positive expression rates of PTEN protein were 47.4% (9/19), 20.0% (2/10) and 92.3% (12/13) in mucinous, endometrioid and the other variants of cervical adenocarcinoma, respectively. Mutation rates at exon 5 and exon 8 of PTEN gene were 19.0% (8/42), 45.0% (9/20) and 0 in cervical adenocarcinoma, cervical glandular intraepithelial neoplasia and normal cervix tissue, respectively. There were significant differences among 3 groups (chi(2) = 4.29, chi(2) = 12.70; P < 0.05). The mutation rates were 21.1% (4/19) and 40.0% (4/10) in mucinous and endometrioid variants of cervical adenocarcinoma, respectively. There was no mutation at exons 5 and 8 of PTEN gene detected in other variants of cervical adenocarcinoma.

CONCLUSIONThe development of cervical adenocarcionomas is correlated with the mutation and absence of the protein expression of PTEN, likely in the early phase of their carcinogenesis.

Adenocarcinoma ; genetics ; metabolism ; Adenocarcinoma, Mucinous ; genetics ; metabolism ; Carcinoma, Endometrioid ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; genetics ; metabolism ; Cervix Uteri ; metabolism ; Exons ; Female ; Humans ; Mutation ; PTEN Phosphohydrolase ; genetics ; metabolism ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Uterine Cervical Neoplasms ; genetics ; metabolism

BACKGROUND: m6A RNA methylation modification plays an important role in the occurrence and progression of lung cancer and regulates tumor immunity. Current studies mostly focus on the differential expression of some specific m6A effectors and infiltrating immune cell. m6A methylation modification is the result of mutual adjustment and balance between effectors, and changes in the expression of one or two effectors are far from enough to reflect the panorama of m6A methylation. The role of m6A in the immune microenvironment of lung adenocarcinoma (LUAD) is still poorly understood. The aim of this study is to investigate the effect of different m6A modification patterns in immune microenvironment of LUAD. METHODS: LUAD data was obtained from The Cancer Genome Atlas (TCGA), University of California Santa Cruz Xena (UCSC Xena) and Gene Expression Omnibus (GEO) databases. Gene mutation, differential expression and survival analysis were performed for 24 m6A effectors. The m6A modification pattern was constructed by unsupervised clustering method, and the m6A clusters survival analysis, gene set variation analysis, immune score and immune cell infiltration analysis were performed. The association between LRPPRC protein expression levels and infiltration of CD8+ cytotoxic T lymphocytes and CD68+ macrophages in the tumor microenvironment was validated by immunohistochemistry in LUAD tissue microarray with 68 cases. RESULTS: The mutations of m6A effector were found in 150 of 567 LUAD cases with a frequency of 26.46%. 6 readers and 3 writers were significantly up regulated in LUAD tissues compared with normal tissues. IGF2BP1 and HNRNPC are the independent risk factors for prognosis of LUAD. Abundant cross-talks among writers, erasers and readers were demonstrated. Three m6A modification patterns with different immune cell infiltration characteristics and clinical prognosis were established. Among m6A effectors, LRPPRC was found to be inversely associated with the infiltration of CD8+ cytotoxic T lymphocytes and CD68+ macrophages, and was validated in 68 LUAD tissues. CONCLUSIONS m6A modification patterns play non-negligible roles in regulating the immune microenvironment. LRPPRC has potential to be a new biomarker for checkpoint inhibitor immunotherapy.
Adenocarcinoma/genetics* ; Adenocarcinoma of Lung/pathology* ; Adenosine/metabolism* ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms/pathology* ; Methylation ; Tumor Microenvironment/genetics*

To explore the role of the Chk2 protein expression and DNA double strand breaks (DSBs) repair in low dose hyper-radiosensitivity (HRS)/increased radioresistance (IRR) of non-small cell lung cancer, A549 cells were subjected to irradiation at the dosage ranging from 0.05-2 Gy. Clonogenic survival was measured by using fluorescence-activated cell sorting (FACS) plating technique. Percentage of cells in M-phase after low doses of X-irradiation was evaluated by phospho-histone H3-FITC/PI and Western blotting was used to detect protein expression of Chk2 and phospo-Chk2. DNA DSBs repair efficiency was also measured by induction and persistence of γ-H2AX. The results showed that the killing ability of irradiation with A549 cells increased at low conditioning dose below 0.3 Gy. Within the dose of 0.3 to 0.5 Gy, A549 cells showed a certain extent of radiation resistance. And when the dose was more than 0.5 Gy, survival fraction exhibited a negative correlation with the dosage. There was no difference between the 0.1 or 0.2 Gy dosage groups and the un-irradiated group in terms of the percentage of cells in M phase. But in the high dosage group (0.3-1.0 Gy), the percentage of cells in M phase was decreased markedly. In addition, the percentage of cells in M phase began to decrease two hours after irradiation. One hour after irradiation, there was no conspicuous activation of Chk2 kinase in 0.1 or 0.2 Gy group, but when the irradiation dose reached 0.3 Gy or higher, Chk2 kinase started to be activated and the activation level showed no significant difference among high dosage groups (0.4, 0.5, 1.0 Gy). Within 1 to 6 h, the DNA DSBs repair efficiency was decreased at 0.2 Gy but increased at 0.5 Gy and 1.0 Gy, which was in line with Chk2 activation. We are led to conclude that the mechanism of HRS/IRR in A549 cell line was probably due to early G(2)/M checkpoint arrest and enhanced DNA DSBs repair. In this regard, Chk2 activation plays a key role in G(2)/M checkpoint activation.
Adenocarcinoma ; genetics ; metabolism ; Cell Line, Tumor ; Checkpoint Kinase 2 ; genetics ; metabolism ; DNA Damage ; genetics ; DNA Repair ; genetics ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Radiation Tolerance ; genetics

OBJECTIVE: To explore the expression of fibronectin type Ⅲ domain containing 1(FNDC1) protein in lung adenocarcinoma and its prognostic significance. METHODS: The expression of FNDC1 in lung adenocarcinoma was predicted by analysis of data from GEO database and GEPIA, and the results were verified by immunohistochemical staining in 92 pairs of clinical specimens of lung adenocarcinoma and adjacent tissues.We further analyzed the correlation of FNDC1 expression with the clinicopathological features of the patients, and evaluated its prognostic value using Cox survival analysis. RESULTS: Analysis of the data form GEO database and GEPIA showed a significantly higher expression level of FNDC1 in lung adenocarcinoma than in matched normal tissues (P < 0.05).Kaplan-Meier survival analysis suggested that a high expression of FNDC1 protein was associated with a significantly shorter overall survival time of the patients (P < 0.05).Immunohistochemistry of the clinical specimens also showed a significantly higher protein expression of FNDC1 in lung adenocarcinoma tissues than in paired adjacent tissues (P < 0.001).A high expression of FNDC1 protein was significantly correlated with advanced clinical stage, T stage and N stage (P < 0.05).Cox univariate and multivariate regression survival analysis indicated that an increased expression of FNDC1 was an independent risk factor for poor prognosis of the patients with lung adenocarcinoma (P < 0.05). CONCLUSION FNDC1 protein is highly expressed in patients with lung adenocarcinoma and in closely related with the occurrence, progression and prognosis of the tumor, suggesting the value of FNDC1 protein as a potential biomarker for assessment of the survival and prognosis of patients with lung adenocarcinoma.
Adenocarcinoma of Lung/metabolism* ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms/metabolism* ; Neoplasm Proteins/genetics* ; Prognosis ; Proteins

Objective Many studies have revealed the crucial roles of miRNA in multiple human cancers, including lung adenocarcinoma (LUAD). In this study, we sought to explore new miRNA-mRNA pairs that are associated with LUAD prognosis. Methods A novel miRNA-mRNA regulatory network associated with prognosis in LUAD was identified and validated using the bioinformatic tools including OncomiR database, StarBase, miRnet, GEPIA2, UALCAN. Results Twenty key miRNAs were compiled after the analysis of the expression and prognostic value in OncomiR and StarBase. Targeted mRNAs of these key miRNAs were predicted in miRnet, and the resulting mRNAs were also analyzed for their prognostic values and expression patterns in GEPIA2 and UALCAN, respectively. Further expression correlation analysis was performed in StarBase. Subsequently, a new miRNA-mRNA network was built, of which each RNA pair showed negative expression correlation, opposite expression pattern, and prognostic value. Protein-protein interaction network was under construction for the mRNAs, and 19 hub genes were determined. Enrichment analysis showed that "Cell Cycle, Mitotic" was the most significantly enriched term. Then, a miRNA-hub gene sub-network was built. We selected and validated the regulatory relationship of some miRNA-hub pairs, including hsa-miR-1976/RFC2, hsa-let-7c-5p/RFC2, hsa-let-7c-5p/ESPL1, hsa-let-7c-5p/CDC25A, and hsa-miR-101-3p/KIF2C. Moreover, over-expression of hsa-miR-1976 and hsa-let-7c-5p resulted in significant cell cycle arrest. Conclusions Our results determined new prognosis-associated miRNA-mRNA pairs and might shed further light on the mechanism via which miRNA-mRNA network influences prognosis in LUAD.
Adenocarcinoma of Lung/genetics* ; Humans ; Lung Neoplasms/pathology* ; MicroRNAs/metabolism* ; Prognosis ; RNA, Messenger/metabolism*

Adenocarcinoma, Follicular ; genetics ; metabolism ; pathology ; Biomarkers, Tumor ; genetics ; Diagnosis, Differential ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; ras Proteins ; biosynthesis ; genetics

OBJECTIVEThis study aimed to investigate the effects of gene polymorphism of heat shock protein 70-2 (HSP 70-2) 1267A/G on the mRNA level HSP 70-2 mRNA and the protein level HSP 70 in human lung cancer.

METHODSForty six lung cancer patients diagnosed histopathologically between February and August 2008 from a hospital in zhengzhou were enrolled as the subjects in this study. Gene polymorphism of HSP 70-2 1276A/G in 46 patients with lung cancer was detected by PCR-RFLP. The mRNA levels of HSP 70-2 mRNA and the protein levels of HSP 70 in lung tissue and para-cancerous tissues of these subjects were determined by RT-PCR and Western blotting respectively.

RESULTSThe expression levels of HSP 70-2 mRNA (1.02 ± 0.30) and HSP 70 protein (0.44 ± 0.12) in the lung cancer tissues was significantly higher than that in para-cancerous tissues (0.19 ± 0.04, 0.12 ± 0.02). The relative levels of HSP 70-2 mRNA in the subjects with AA genotype (1.32 ± 0.22) were significantly higher than the patients with AG genotype or GG genotype (0.95 ± 0.17, 0.70 ± 0.16) at the site of 1267 (A/G) (P < 0.01); however, the relative protein levels of HSP 70 were 0.47 ± 0.13 (AA genotype), 0.42 ± 0.11 (AG genotype), 0.45 ± 0.11 (GG genotype), respectively, which showed no statistically significant difference (P > 0.05).

CONCLUSIONThe polymorphism of HSP 70-2 1267 (A/G) is highly associated with the transcription level of HSP 70-2 mRNA, but not with the expression level of HSP 70 protein.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Female ; Genotype ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Lung ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Neoplasm Staging ; Polymorphism, Single Nucleotide ; RNA, Messenger ; genetics

A mammalian expression plasmid pcDNA3. 1-hCC10 was constructed and identified, then CC10 protein expression in A549 lung cancer cell line was detected. A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by using RT-PCR and cloned into expression plasmid cDNA3. 1, and the recombinant plasmid was identified by employing double digestion restriction enzymes Hind III and BamH I and the cDNA sequence was assayed by the Sanger dideoxy-mediated chain termination method. The segment was then transfected into the A549 lung cancer cell line. The protein expression of CC10 was detected by immunofluorescence and Western blot. Our results showed that the cDNA fragment included the entire coding region (273 bp). The recombinant eukaryotic cell expression vector of pcDNA3. 1-hCC10 was successfully constructed, and the sequence of the insert was identical to the published sequence. A549 cells line transfected with the pcDNA3. 1-hCC10 expressed high level of CC10 protein. The recombinant plasmid cDNA3. 1-hCC10 may serve as an effective tool for the study of tumorogenesis and tumor treatment.
Adenocarcinoma/*metabolism ; Adenocarcinoma/pathology ; Cell Line, Tumor ; Genetic Vectors ; Lung Neoplasms/*metabolism ; Lung Neoplasms/pathology ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Transfection ; Uteroglobin/biosynthesis ; Uteroglobin/*genetics