For first-line non-small-cell lung cancer（NSCLC） therapy,detecting mutation status of the epidermal growth factor receptor（EGFR） gene constitutes a prudent test to identify patients who are most likely to benefit ...For first-line non-small-cell lung cancer（NSCLC） therapy,detecting mutation status of the epidermal growth factor receptor（EGFR） gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor（TKI） therapy.Now,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded（FFPE） tissues.DNA extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction（PCR） are two key steps for detecting EGFR gene mutation.We showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR gene.Extracting DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100（pH=10.0） was performed by heating at 95 °C for 30 min.Meanwhile,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for comparison.DNA extracted products were used as template for amplifying the exons 18,19,20 and 21 of EGFR by PCR for different amplified fragments.Results show that DNA fragment size extracted from FFPE tissues with 1% Triton X was about 250―500 base pairs（bp）.However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands bp.The DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial kit.For about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial kit.However,for about 200 bp fragment.This simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.展开更多
Background:To investigate the mutation types and mutation rate of the epidermal growth factor receptor(EGFR)gene in patients with lung adenocarcinoma and the clinical features of lung adenocarcinoma with EGFR gene mut...Background:To investigate the mutation types and mutation rate of the epidermal growth factor receptor(EGFR)gene in patients with lung adenocarcinoma and the clinical features of lung adenocarcinoma with EGFR gene mutations in Karamay,Xinjiang,China.Methods:Paraffin-embedded tissue samples of adenocarcinoma patients were collected in the Karamay Central Hospital from March 2016 to June 2019,and mutations in exon 18–21 of the EGFR gene were detected by the allele-specific amplification polymerase chain reaction(Amplification RefractoryMutation System–PCR)method.The relationships between themutation types,mutation incidence,and clinical features were analyzed.Results:Of the 170 patients with lung adenocarcinoma,83 had EGFR mutations.The total mutation rate of EGFR in patients with lung adenocarcinoma was 48.8%,which included mutations in exons 18(1.2%[2/170]),19(19.4%[33/170]),20(2.4%[4/170]),and 21(20.6%[35/170]).Intriguingly,there was a case with 9 mutations in exons 20 and 21.The mutations in exon 19 of EGFR resulted in the deletion of codons 746 to 750.The main mutation in exon 21 was L858R(91.4%[32/35]).There was no significant difference in exons 19 and 21 mutation rates(P>0.05).The mutation rate of EGFR in female patients was significantly higher than that in male patients(P<0.05)but had no correlation with the age,smoking status,and clinical stage of patients with non–small cell lung cancer(P>0.05).The EGFR mutation rate may be related to the degree of tumor differentiation.Conclusions:Among patients with lung adenocarcinoma in Kelamayi(city in Xinjiang),EGFR mutations were more frequently detected in female patients,and the main sites of mutations were exons 19 and 21.展开更多
Objective：The epidermal growth factor receptor （EGFR） inhibitors monoclonal antibodies （MoAbs） have already shown the therapeutic effectiveness in patients with metastatic colorectal cancer （mCRC）.But many pati...Objective：The epidermal growth factor receptor （EGFR） inhibitors monoclonal antibodies （MoAbs） have already shown the therapeutic effectiveness in patients with metastatic colorectal cancer （mCRC）.But many patients resist to the treatment.The aim of this meta-analysis was to assess EGFR gene copy number （GCN） as a candidate predictive biomarker for resistance to anti-EGFR MoAbs in mCRC treatment.Methods：Systematic computerized searches of the PubMed,EMBase and Cochrane Library were performed.The primary endpoint was objective response rate （ORR）.The second endpoints included progression-free survival （PFS）,and overall survival （OS）.The pooled odd ratio （OR） and pooled sensitivity,specificity,and summary receiver operator characteristic （SROC） for ORR were estimated.The pooled hazard ratios （HR） for PFS and OS were also calculated.Results：Fourteen studies with 1,021 patients were included.Increased EGFR GCN was associated with increased ORR （OR=6.905; 95％ CI：4.489-10.620）.It was also found in wild-type KRAS mCRC patients,with the pooled OR of 8.133 （95 ％ CI：4.316-15.326）.GCN has medium value for predicting ORR,with the pooled sensitivity of 0.79 （95％ CI：0.73-0.84）,the pooled specificity of 0.59 （95％ CI：0.55-0.62）.In wildtype KRAS mCRC patients,the sensitivity and the specificity were 0.80 （95％ CI：0.70-0.87） and 0.60 （95％CI：0.53-0.66）,respectively.Increased EGFR GCN was associated with increased PFS （HR=0.557; 95％ CI：0.382-0.732） and OS （HR=0.579; 95％ CI：0.422-0.737）.Conclusions：This meta-analysis suggests that EGFR GCN represents a predictive biomarker for tumor response in mCRC patients treated with MoAbs regardless of KRAS mutation.mCRC patients with increased EGFR GCN are more likely to have a better response,PFS,and OS when treated with cetuximab or panitumumab.展开更多
Epidermal growth factor receptor （EGFR） is frequently overexpressed in non-small-cell lung cancer （NSCLC） and plays a key role in tumorigenesis.1 Small molecule tyrosine kinase inhibitors （TKIs）,such as gefitini...Epidermal growth factor receptor （EGFR） is frequently overexpressed in non-small-cell lung cancer （NSCLC） and plays a key role in tumorigenesis.1 Small molecule tyrosine kinase inhibitors （TKIs）,such as gefitinib,erlotinib,and icotinib,which can inhibit receptor tyrosine kinase activity of EGFR become clinically available for the treatment of non-small-cell lung cancer （NSCLC）.2 NSCLC patients with EGFR mutation have experienced a marked response to EGFR-TKIs therapy.3 Detection of mutations of the EGFR gene is critical for predicting the response to therapy with TKIs.4展开更多
To further illustrate the roles of p53 gene, epidermal growth factor receptor (EGFR) gene and loss of heterozygosity (LOH) on chromosome 10 and 17?p in human glioma progression Methods p53 mutations were scann...To further illustrate the roles of p53 gene, epidermal growth factor receptor (EGFR) gene and loss of heterozygosity (LOH) on chromosome 10 and 17?p in human glioma progression Methods p53 mutations were scanned in 50 gliomas with various malignant grades using the polymerase chain reaction single strand conformation polymorphism (PCR SSCP) assay, and were confirmed by direct sequencing LOH for chromosome 10, 17?p and amplification of the EGFR gene were also assessed using Southern blot analysis Results p53 mutations were found in 9 of 17 high grade astrocytomas (53%), 1 of 15 low grade astrocytomas (7%), and the only subject of eppendymoblastoma but in none of the 10 medulloblastomas and 7 eppendymomas The majority of gliomas (38/50) analyzed here retained both 17?p alleles The frequency of p53 mutations was 13% in this group of tumors and increased to 50% (6/12) in tumors with one 17?p allele ( P <0 025) LOH on chromosome 10 was found in 35% (6/17) of high grade astrocytomas, in 10% (1/10) of medulloblastomas, but in 0% of low grade gliomas EGFR gene amplification was found in 9 high grade gliomas, 60% (6/9) of which also presented LOH for chromosome 10 Conclusions These results indicate that p53 inactivation is a common genetic event in astrocytoma progression that may be more strongly associated with the progression of astrocytomas than with their origin Absence of p53 mutations in 50% of the tumors with one 17?p allele suggests that a tumor suppressor gene other than p53 may be located on chromosome 17?p and involved in progression to malignancy of some gliomas The loss of alleles on chromosome 10 and the amplification of the EGFR gene appear to be restricted to high grade tumors, suggesting that these events may be related to tumor progression rather than initiation展开更多
基金Supported by the Jilin Science & Technology Development Plan,China(No.201201060)the Scientific Research Foundation of Jilin Province,China(No.20100942)+1 种基金the Fund of Developing and Reforming Community of Jilin Province,China(No.2010-1928)the Health Scientific Research Foundation of Jilin Province,China(Nos.2009z081,2010Z083)
文摘For first-line non-small-cell lung cancer（NSCLC） therapy,detecting mutation status of the epidermal growth factor receptor（EGFR） gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor（TKI） therapy.Now,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded（FFPE） tissues.DNA extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction（PCR） are two key steps for detecting EGFR gene mutation.We showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR gene.Extracting DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100（pH=10.0） was performed by heating at 95 °C for 30 min.Meanwhile,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for comparison.DNA extracted products were used as template for amplifying the exons 18,19,20 and 21 of EGFR by PCR for different amplified fragments.Results show that DNA fragment size extracted from FFPE tissues with 1% Triton X was about 250―500 base pairs（bp）.However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands bp.The DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial kit.For about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial kit.However,for about 200 bp fragment.This simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.
基金supported by a grant fromthe Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2021D01A24).
文摘Background:To investigate the mutation types and mutation rate of the epidermal growth factor receptor(EGFR)gene in patients with lung adenocarcinoma and the clinical features of lung adenocarcinoma with EGFR gene mutations in Karamay,Xinjiang,China.Methods:Paraffin-embedded tissue samples of adenocarcinoma patients were collected in the Karamay Central Hospital from March 2016 to June 2019,and mutations in exon 18–21 of the EGFR gene were detected by the allele-specific amplification polymerase chain reaction(Amplification RefractoryMutation System–PCR)method.The relationships between themutation types,mutation incidence,and clinical features were analyzed.Results:Of the 170 patients with lung adenocarcinoma,83 had EGFR mutations.The total mutation rate of EGFR in patients with lung adenocarcinoma was 48.8%,which included mutations in exons 18(1.2%[2/170]),19(19.4%[33/170]),20(2.4%[4/170]),and 21(20.6%[35/170]).Intriguingly,there was a case with 9 mutations in exons 20 and 21.The mutations in exon 19 of EGFR resulted in the deletion of codons 746 to 750.The main mutation in exon 21 was L858R(91.4%[32/35]).There was no significant difference in exons 19 and 21 mutation rates(P>0.05).The mutation rate of EGFR in female patients was significantly higher than that in male patients(P<0.05)but had no correlation with the age,smoking status,and clinical stage of patients with non–small cell lung cancer(P>0.05).The EGFR mutation rate may be related to the degree of tumor differentiation.Conclusions:Among patients with lung adenocarcinoma in Kelamayi(city in Xinjiang),EGFR mutations were more frequently detected in female patients,and the main sites of mutations were exons 19 and 21.
文摘Objective：The epidermal growth factor receptor （EGFR） inhibitors monoclonal antibodies （MoAbs） have already shown the therapeutic effectiveness in patients with metastatic colorectal cancer （mCRC）.But many patients resist to the treatment.The aim of this meta-analysis was to assess EGFR gene copy number （GCN） as a candidate predictive biomarker for resistance to anti-EGFR MoAbs in mCRC treatment.Methods：Systematic computerized searches of the PubMed,EMBase and Cochrane Library were performed.The primary endpoint was objective response rate （ORR）.The second endpoints included progression-free survival （PFS）,and overall survival （OS）.The pooled odd ratio （OR） and pooled sensitivity,specificity,and summary receiver operator characteristic （SROC） for ORR were estimated.The pooled hazard ratios （HR） for PFS and OS were also calculated.Results：Fourteen studies with 1,021 patients were included.Increased EGFR GCN was associated with increased ORR （OR=6.905; 95％ CI：4.489-10.620）.It was also found in wild-type KRAS mCRC patients,with the pooled OR of 8.133 （95 ％ CI：4.316-15.326）.GCN has medium value for predicting ORR,with the pooled sensitivity of 0.79 （95％ CI：0.73-0.84）,the pooled specificity of 0.59 （95％ CI：0.55-0.62）.In wildtype KRAS mCRC patients,the sensitivity and the specificity were 0.80 （95％ CI：0.70-0.87） and 0.60 （95％CI：0.53-0.66）,respectively.Increased EGFR GCN was associated with increased PFS （HR=0.557; 95％ CI：0.382-0.732） and OS （HR=0.579; 95％ CI：0.422-0.737）.Conclusions：This meta-analysis suggests that EGFR GCN represents a predictive biomarker for tumor response in mCRC patients treated with MoAbs regardless of KRAS mutation.mCRC patients with increased EGFR GCN are more likely to have a better response,PFS,and OS when treated with cetuximab or panitumumab.
文摘Epidermal growth factor receptor （EGFR） is frequently overexpressed in non-small-cell lung cancer （NSCLC） and plays a key role in tumorigenesis.1 Small molecule tyrosine kinase inhibitors （TKIs）,such as gefitinib,erlotinib,and icotinib,which can inhibit receptor tyrosine kinase activity of EGFR become clinically available for the treatment of non-small-cell lung cancer （NSCLC）.2 NSCLC patients with EGFR mutation have experienced a marked response to EGFR-TKIs therapy.3 Detection of mutations of the EGFR gene is critical for predicting the response to therapy with TKIs.4
基金ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina (No 395 70 714 )
文摘To further illustrate the roles of p53 gene, epidermal growth factor receptor (EGFR) gene and loss of heterozygosity (LOH) on chromosome 10 and 17?p in human glioma progression Methods p53 mutations were scanned in 50 gliomas with various malignant grades using the polymerase chain reaction single strand conformation polymorphism (PCR SSCP) assay, and were confirmed by direct sequencing LOH for chromosome 10, 17?p and amplification of the EGFR gene were also assessed using Southern blot analysis Results p53 mutations were found in 9 of 17 high grade astrocytomas (53%), 1 of 15 low grade astrocytomas (7%), and the only subject of eppendymoblastoma but in none of the 10 medulloblastomas and 7 eppendymomas The majority of gliomas (38/50) analyzed here retained both 17?p alleles The frequency of p53 mutations was 13% in this group of tumors and increased to 50% (6/12) in tumors with one 17?p allele ( P <0 025) LOH on chromosome 10 was found in 35% (6/17) of high grade astrocytomas, in 10% (1/10) of medulloblastomas, but in 0% of low grade gliomas EGFR gene amplification was found in 9 high grade gliomas, 60% (6/9) of which also presented LOH for chromosome 10 Conclusions These results indicate that p53 inactivation is a common genetic event in astrocytoma progression that may be more strongly associated with the progression of astrocytomas than with their origin Absence of p53 mutations in 50% of the tumors with one 17?p allele suggests that a tumor suppressor gene other than p53 may be located on chromosome 17?p and involved in progression to malignancy of some gliomas The loss of alleles on chromosome 10 and the amplification of the EGFR gene appear to be restricted to high grade tumors, suggesting that these events may be related to tumor progression rather than initiation
