1.Impact of smoking status and pathologic type on epidermal growth factor receptor mutations in lung cancer.
Yi-Sheng HUANG ; Jin-Ji YANG ; Xu-Chao ZHANG ; Xue-Ning YANG ; Yu-Juan HUANG ; Chong-Rui XU ; Qing ZHOU ; Zhen WANG ; Jian SU ; Yi-Long WU
Chinese Medical Journal 2011;124(16):2457-2460
BACKGROUNDEpidermal growth factor receptor (EGFR) mutations in lung carcinomas can make the disease more responsive to the treatment with tyrosine kinase inhibitors. We aimed to evaluate the prevalence of EGFR mutations in a large series of lung carcinomas.
METHODSWe examined 1195 consecutive lung cancer patients for EGFR mutations in exons 18, 19, and 21 using direct sequencing of polymerase chain reaction products. A detailed smoking history was obtained. Patients were categorized as never smokers (< 100 lifetime cigarettes), former smokers (quit > 1 year ago), or current smokers (quit < 1 year ago).
RESULTSThere were EGFR mutations in 9 (4.5%) of 201 squamous carcinomas, in 1 (2%) of 50 large cell carcinomas, and in 1 (2.3%) of 44 small cell carcinomas that were investigated. Three hundred and twenty-seven mutations were found in the series of 858 adenocarcinomas (38.1%). Among 858 lung adenocarcinomas, we detected EGFR mutations in 250 (48.6%) of 514 never smokers, 39 (33.9%) of 115 former smokers, and 38 (16.6%) of 229 current smokers. Significantly fewer EGFR mutations were found in people who smoked for more than 15 pack-years (P = 0.0002) or stopped smoking less than 15 years ago (P = 0.033) compared with individuals who never smoked.
CONCLUSIONSAdenocarcinoma is the most frequent EGFR mutation pathologic type in lung cancer. The likelihood of EGFR mutations in exons 18, 19 and 21 decreases as the number of pack-years increases. Mutations were less common in people who smoked for more than 15 pack-years or who stopped smoking cigarettes less than 15 years ago. These data can assist clinicians in assessing the likelihood of exons 18, 19, or 21 EGFR mutations in Chinese patients with lung cancer when mutational analysis is not feasible.
Adenocarcinoma ; genetics ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Mutation ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; Smoking ; adverse effects
3.Research on autosomal dominant polycystic kidney disease in China.
Chinese Medical Journal 2006;119(22):1915-1924
OBJECTIVETo review the history and recent development of research on autosomal dominant polycystic kidney disease (ADPKD) in China.
DATA SOURCESBoth Chinese and English literatures were searched in MEDLINE/CD ROM (1979 - 2006) and the Chinese Biomedical Literature Disk (1979 - 2006).
STUDY SELECTIONPublished articles about ADPKD from mainland of China were selected. Data were mainly extracted from 58 articles which are listed in the reference section of this review.
RESULTSSome preliminary reports on cyst decompression surgeries and mutation analysis represent the contribution to the ADPKD research from China in the history. A serial of basic research and clinical studies on ADPKD in recent years also have been summarized. A technique platform for ADPKD research was firstly established. The genomics/proteomics/bioinformatics approach was introduced, which provide a lot of valuable information for understanding the pathogenesis. By denature high performance liquid chromatography (DHPLC) technique the entire PKD1 and PKD2 gene sequence screening system for Chinese Han population has been successfully established. Based on the characteristic data of Chinese patients, an integrated therapy protocol was put forward and won an advantage over the traditional therapy. Some novel experimental studies on therapy also were encouraging.
CONCLUSIONSRemarkable progress of ADPKD research in China have been made recently. Still many works, including the government support, international collaboration and active participation of more Chinese nephrologists, should be enhanced to advance this process in the near future.
Cell Line ; China ; Humans ; Peptide Fragments ; analysis ; Polycystic Kidney, Autosomal Dominant ; diagnosis ; etiology ; genetics ; therapy ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Research ; TRPP Cation Channels ; analysis
4.Expression and identification of the extracellular domain of human epidermal growth factor receptor type III variant.
Xing-mei ZHANG ; Yu-sheng SHI ; Zhong-ying LIU
Journal of Southern Medical University 2008;28(2):151-153
OBJECTIVETo clone the extracellular domain (ECD) of the type III variant of human epidermal growth factor receptor (EGFRvIII) and construct the recombinant expression plasmid.
METHODSA DNA fragment (vIII ECD) encoding the extracellular domain of human EGFRvIII was obtained by PCR, and its T-A was cloned and sequenced. The DNA fragment was then ligated into the GST fusion expression vector to construct the recombination plasmid. After identification with restriction digestion and DNA sequencing, the recombinant plasmid was transformed into E. coli BL21 (DE3) for expression of the recombinant protein. The target protein was identified by SDS-PAGE and Western blotting.
RESULTSThe results of restriction digestion and DNA sequencing confirmed the successful construction of the recombinant plasmid. SDS-PAGE showed that the fusion protein was expressed as inclusion bodies in E. coli BL21 (DE3), and the amount of the fusion protein expressed in the bacteria, after induction for 4 h, accounted for up to 15% of the total bacterial proteins. Western blotting demonstrated that the fusion protein could be recognized by the specific anti-EGFR antibody.
CONCLUSIONWe have successfully constructed the recombinant expression vector of vIII ECD and induced the expression of the fusion protein, which may facilitate functional and immunological studies of EGFRvIII.
Cloning, Molecular ; Escherichia coli ; Genetic Vectors ; Humans ; Plasmids ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Sequence Analysis, DNA
5.Effect of epidermal growth factor receptor on airway remodeling in asthmatic mice and its mechanism.
Chinese Journal of Contemporary Pediatrics 2010;12(2):137-140
OBJECTIVETo explore the relationship of airway remodeling with epidermal growth factor receptor (EGFR) and heparin-binding epidermal growth factor-like growth factor (HB-EGF) levels in asthmatic mice and the effect of EGFR tyrosine kinase inhibitor (AG1478) on airway remodeling.
METHODSTwenty-four male BALB/c mice were randomly divided into three groups: normal control, asthma, AG1478-treated. Mice were sensitized and challenged with ovalbumin (OVA) and a mouse mode1 of asthma was prepared. Collagen deposition was determined in Masson-stained lung sections. Periodic acid Schiff (PAS) staining was used to observe the proliferation of goblet cells. Immunohistochemistry was used to determine the protein expression of HB-EGF. RT-PCR was used to determine the mRNA expression of HB-EGF and EGFR.
RESULTSThe characteristic changes of airway remodeling occurred in the asthma group. The expression of HB-EGF and EGFR in the epithelial cells of bronchi in the asthma group was significantly higher than that in the normal control group. Compared with the asthma group, the AG1478-treated group had decreased inflammation reactions, decreased collagen deposition and proliferation of goblet cells and lower expression of EGFR and HB-EGF.
CONCLUSIONSEGFR tyrosine kinase inhibitor (AG1478) ameliorates the progression of airway remodeling in mice with asthma by inhibitions of EGFR and HB-EGF expression and EGFR signal pathway.
Airway Remodeling ; Animals ; Asthma ; drug therapy ; pathology ; Heparin-binding EGF-like Growth Factor ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; analysis ; genetics ; Male ; Mice ; Quinazolines ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; genetics ; physiology ; Tyrphostins ; therapeutic use
6.Comparison of three methods for detecting epidermal growth factor receptor mutations in plasma DNA samples of Chinese patients with advanced non-small cell lung cancer.
Ling QIN ; Wei ZHONG ; Li ZHANG ; Long-yun LI ; Meng-zhao WANG
Chinese Medical Journal 2011;124(6):887-891
BACKGROUNDEpidermal growth factor receptor (EGFR) mutations can predict tumor response to tyrosine kinase inhibitors (TKIs). Detecting EGFR mutations in plasma DNA samples in patients with advanced non-small cell lung cancer is challenging and promising. We compared three methods for detecting plasma EGFR mutations, including direct DNA sequencing, denaturing high-performance liquid chromatography (DHPLC) and Scorpions Amplification Refractory Mutation System (Scorpions ARMS).
METHODSPlasma DNA samples from 73 patients with stage IIIB to IV adenocarcinoma were analyzed for EGFR mutations in exons 19 (deletion mutation) and 21 (L858R mutation) using direct DNA sequencing, DHPLC and Scorpions ARMS. Sensitivities of the three methods were compared and the relationship between EGFR mutations and patients' survival was analyzed.
RESULTSIn 73 patients, we detected EGFR mutations in 5 samples (6.9%) by direct DNA sequencing, in 22 samples (30.1%) by DHPLC, and in 28 samples (38.4%) by Scorpions ARMS. EGFR mutations were found in 13 samples in exon 19 and in 9 samples in exon 21 by DHPLC, while we found mutations in 15 samples in exon 19 and in 13 samples in exon 21 by Scorpions ARMS. Among the 73 patients, there was 90.4% concordance between DHPLC and Scorpions ARMS (66/73, κ = 0.79, P = 0.07). Of the 73 patients, 46 patients were treated with gefitinib, including 18 patients with mutations and 28 patients without mutations as determined by Scorpions ARMS. The 18 patients with mutations had a significantly longer progression-free survival (PFS) time (median PFS was 21.0 months) than the 28 patients without mutations (median PFS was 7.0 months) (P = 0.022).
CONCLUSIONSAmong the three methods for detecting EGFR mutations in plasma DNA samples of patients with advanced lung adenocarcinoma, direct gene sequencing had the lowest sensitivity, while Scorpion ARMS showed the highest mutation detecting capability. DHPLC is slightly less sensitive than Scorpion ARMS. EGFR mutations in exons 19 (deletion mutation) and 21 (L858R mutation) predict a longer PFS.
Asian Continental Ancestry Group ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; methods ; Exons ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; Reproducibility of Results ; Sequence Analysis, DNA
7.Epithelial growth factor receptor (EGFR) exon double-sequencing analysis in NSClC.
Ren-sheng LAI ; Ling XIE ; Long-shu SHEN ; Chang-le ZHU ; Jun QIAN
Chinese Journal of Oncology 2006;28(8):599-602
OBJECTIVETo study the mutation patterns of epithelial growth factor receptor (EGFR) exon 18, 19 and 21 in Chinese non-small-cell lung cancers (NSCLC).
METHODSSomatic mutation in samples of 32 cases without Iressa-treatment were compared with that in 10 volunteers blood control. The mutations were identified for the forward and reverse sequence chains for the tyrosine kinase domain of the EGFR gene, followed by DNA template abstraction and Touchdown PCR.
RESULTSNine types of mutation were found in sequences of 7 cases among the 32 non-small cell lung carcinoma tissues, namely, five reported mutation within exon 19, and two new heterozygous mutations, L833V and H835L within exon 21, and two intron polymorphism. These results showed a mutation rate of 9/32 (28.1%) in Chinese with NSCLC, and of 31.6% in lung adenocarcinomas.
CONCLUSIONEGFR mutation rate in Chinese with NSCLC is consistent with those of Asian women reported in the literature but new mutation points in Chinese were presented as L833V and H835L. The mutation rate is in concordance with release rate of NSCLC obtained by Gefitinib treatment in Chinese.
Adenocarcinoma ; genetics ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Carcinoma, Non-Small-Cell Lung ; ethnology ; genetics ; China ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Humans ; Lung Neoplasms ; ethnology ; genetics ; Male ; Middle Aged ; Mutation ; Receptor, Epidermal Growth Factor ; genetics
8.p53 Mutation and Epidermal Growth Factor Receptor Overexpression in Glioblastoma.
Kyung Sik YOON ; Min Cheol LEE ; Sam Suk KANG ; Jae Hyoo KIM ; Shin JUNG ; Yoon Ji KIM ; Jae Hyuk LEE ; Kyu Yoon AHN ; Ji Shin LEE ; Jong Yoon CHEON
Journal of Korean Medical Science 2001;16(4):481-488
Recent molecular studies indicate two different genetic pathways leading to the development of glioblastoma; final progression of astrocytoma and de novo formation. To define the mutual relationships of cytogenetic changes in the pathogenesis of glioblastoma, molecular histopathologic alterations of p53 and epidermal growth factor receptor (EGFR) were evaluated by single stranded conformational polymorphion, reverse transcriptase-polymerase chain reaction and immunohistochemical stains in 15 primary and 21 secondary glioblastomas. Mutations in p53 gene and positive immunoreactivity to p53 protein (DO1) were more prevalent in secondary glioblastomas than in primary glioblastomas. A correlation between p53 mutations and p53 immunopositivities in glioblastomas was observed in 83.3% of the cases. All cases with positive p53 immunoreactivities showed p53 mutations; however, 13.9% of glioblastomas with p53 immuno-positivities lacked the relevant mutations. EGFR amplifications were detected in 73.3% of primary glioblastomas and 9.5% of secondary glioblastomas (p<0.001). The concurrence of p53 mutation and EGFR amplification was revealed in only 2 out of 15 primary glioblastomas and none among the secondary glioblastomas. Immunoreactivities for EGFR were noted in 66.7% of primary glioblastomas and in 9.5% of secondary glioblastomas (p<0.001). A correlation between EGFR amplification and EGFR immunopositivity in glioblastomas was observed in 91.7% of the cases. These data indicate that EGFR amplification and p53 mutations are two independent genetic events in the development of glioblastomas.
Adolescence
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Adult
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Brain Neoplasms/*genetics/metabolism
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Female
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*Genes, p53
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Glioblastoma/*genetics/metabolism
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Human
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Immunohistochemistry
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Loss of Heterozygosity
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Male
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Middle Age
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*Mutation
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Protein p53/analysis
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Receptor, Epidermal Growth Factor/analysis/*genetics
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Reverse Transcriptase Polymerase Chain Reaction
9.The construction of recombinant adenovirus expressing bifunctional fusion protein sCAR-EGF and the detection of its activity.
Peng-Kang REN ; Feng WANG ; Hui-Ming LI ; Zong-Hai LI ; Qian HUANG
Chinese Journal of Biotechnology 2006;22(5):713-719
To improve the targeting of adenovirus vector for gene therapy, a fusion gene sCAR-EGF, in which epidermal growth factor gene was fused to the 3' end of extracellular Coxsackie virus-adenovirus receptor gene, was constructed and cloned into shuttle plasmid pDC315 to obtain a recombinant plasmid pDC315-sCAR-EGF. With the AdMax system, AD-293 cells were co-transfected with pDC315-sCAR-EGF and adenovirus genomic plasmid pBHGloxdeltaE13cre. Through high efficiency site specific recombination, a replication-defective adenovirus Ad5-CMV-sCAR-EGF was constructed. The recombinant adenovirus was analyzed by PCR and Western blotting, the results indicated that Ad5-CMV-sCAR-EGF contained the fusion gene sCAR-EGF, and the adenovirus infected cells was induced to produce and secrete the fusion protein into the supernatant. We have demonstrated that the fusion protein sCAR-EGF is helpful for elevating the infection efficiency of Ad5-CMV-luc with the reporter gene in vitro, which providing a new approach to the gene therapy for tumors overexpressing EGFR.
Adenoviridae
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genetics
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Cell Line
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Coxsackie and Adenovirus Receptor-Like Membrane Protein
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Enzyme-Linked Immunosorbent Assay
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Epidermal Growth Factor
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analysis
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genetics
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Genetic Therapy
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Humans
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Neoplasms
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therapy
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Polymerase Chain Reaction
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Receptors, Virus
;
analysis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
10.Clinicopathological characteristics and prognosis of patients with adenosquamous lung carcinoma.
Ye GUO ; Lin JIA ; Guo-Guang SHAO ; Hong-Wei SUN ; Xing-Xing WANG ; Guan-Jun WANG ; Ke-Wei MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(3):350-355
This study was aimed to characterize clinicopathological features and prognosis of patients with adenosquamous lung carcinoma (ASC). Among the 2531 patients with lung cancer who underwent surgery between January 2000 and June 2012 in our hospital, 59 were histologically diagnosed as having ASC. The clinicopathological features and follow-up data of ASC patients were collected and analyzed statistically. Superior lobectomy was accomplished in 40 patients, middle and inferior lobectomy in 3, lobectomy plus partial resection of contralateral lung in 5, partial lung resection in 4, and pneumonectomy in 7. Moreover, 22 cases were found to be adenocarcinoma-predominant, and 18 to be squamous cell carcinoma-predominant. The median survival time was 13.6 months, and the 1-, 3-, and 5-year survival rates were 59.9%, 36.4% and 31.2%, respectively. Of the 52 cases with tissue specimens available, 11 had an EGFR mutation (21.2%) and 2 had a KRAS mutation (3.8%). Multivariate analysis showed that histology subtype, pleural invasion, TNM stage, and postoperative treatment were all independent prognostic factors. The data from the current study demonstrated that SCC-predominant histology represents a better prognosis of ASC. Histology subtype, pleural invasion, TNM stage, and postoperative treatment are independent prognostic factors for ASC and adjuvant therapy may help control the disease.
Adult
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Aged
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Carcinoma, Adenosquamous
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genetics
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pathology
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surgery
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Diagnosis, Differential
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Female
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Humans
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Lung Neoplasms
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genetics
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pathology
;
surgery
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Male
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Middle Aged
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Mutation
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Prognosis
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Proto-Oncogene Proteins p21(ras)
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genetics
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Receptor, Epidermal Growth Factor
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genetics
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Retrospective Studies
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Survival Analysis