Progress of Immunotherapy in EGFR-mutated Advanced Non-small Cell Lung Cancer / 中国肺癌杂志

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) are currently the first-line standard of care for patients with non-small cell lung cancer (NSCLC) that harbor EGFR mutations. Nevertheless, resistance to EGFR-TKIs is inevitable. In recent years, although immune checkpoint inhibitors (ICIs) have significantly shifted the treatment paradigm in advanced NSCLC without driver mutation, clinical benefits of these agents are limited in patients with EGFR-mutated NSCLC. Compared with wild-type tumors, tumors with EGFR mutations show more heterogeneity in the expression level of programmed cell death ligand 1 (PD-L1), tumor mutational burden (TMB), and other tumor microenvironment (TME) characteristics. Whether ICIs are suitable for NSCLC patients with EGFR mutations is still worth exploring. In this review, we summarized the clinical data with regard to the efficacy of ICIs in patients with EGFR-mutated NSCLC and deciphered the unique TME in EGFR-mutated NSCLC.
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EGFR gene mutations and environmental exposure factors in patients with non-small cell lung cancer in Bazhong City / 公共卫生与预防医学

Objective To analyze the EGFR gene mutations and environmental exposure factors in patients with non-small cell lung cancer (NSCLC) in Bazhong City, and to provide a theoretical basis for the diagnosis and treatment of NSCLC patients. Methods A total of 356 NSCLC patients admitted to Bazhong Hospital from 2019 to 2020 were selected. All patients underwent EGFR gene detection and were divided into mutant group (n=171) and wild-type group (n=185) according to EGFR gene mutation. Environmental exposure data of patients were collected, including smoking status, smoking index, frequent frying of food, etc. Univariate analysis and logistic regression were used to analyze the environmental risk factors of EGFR gene mutations in NSCLC patients. Results A total of 171 EGFR gene mutations were detected in 356 NSCLC patients, and the mutation rate was 48.03%. The mutation rate of EGFR gene in females was significantly higher than that in males (P0.05). The mutation rate of EGFR gene in patients with adenocarcinoma was significantly higher than that in patients without adenocarcinoma (P0.05). Among the 356 NSCLC patients, there were 171 cases with EGFR gene mutations (48.03%), including 335 single mutations, 181 exon 19 mutations, 129 exon 21 L858R mutations, 12 exon 21 L861Q mutations, 8 exon 20 insertion mutations, and 5 Exon 18 mutations. There were 18 cases carrying double mutations and 3 cases carrying triple mutations. There were significant differences between the two groups in smoking status, smoking index, use of coal stove, use of smoke extraction equipment, cooking fumes, fried food intake, and family history of cancer (P<0.05). Non-smoking (OR=3.19), not using smoke exhaust equipment (OR=3.58), and using coal stove (OR=2.19) were the environmental exposure factors of EGFR mutation in NSCLC patients (P<0.05). Conclusion The EGFR gene mutation rate is high in NSCLC patients in Bazhong City, and most of them are female non-smoking patients. EGFR gene detection should be performed in NSCLS patients without smoke exhaust equipment and using coal stoves to improve the detection rate of EGFR mutation.

Clinicopathological Features and Prognosis of Patients Newly Diagnosed With Lung Adenocarcinoma With Both EGFR Mutation and C-MET Amplification / 中国医学科学院学报

Objective To explore the clinicopathological features and prognosis of the patients newly diagnosed with lung adenocarcinoma with both EGFR mutation and C-MET amplification.Methods The pathological sections were reviewed.EGFR mutation was detected by amplification refractory mutation system-quantitative real-time polymerase chain reaction,and C-MET amplification by fluorescence in situ hybridization.The clinicopathological features and survival data of the patients newly diagnosed with lung adenocarcinoma with both EGFR mutation and C-MET amplification were analyzed retrospectively.Results In 11 cases of EGFR mutation combined with C-MET amplification,complex glands and solid high-grade components were observed under a microscope in 10 cases except for one case with a cell block,the tissue structure of which was difficult to be evaluated.The incidence of lung adenocarcinoma in the patients with EGFR mutation combined with C-MET amplification at clinical stage Ⅳ was higher than that in the EGFR mutation or C-MET amplification group (all P<0.001),whereas the difference was not statistically significant between the EGFR mutation group and C-MET amplification group at each clinical stage (all P>0.05).There was no significant difference in the trend of survival rate between EGFR gene group and C-MET amplification group (χ2=0.042,P=0.838),while the survival of the patients with EGFR mutation combined with C-MET amplification was worse than that of the patients with EGFR mutation (χ2=246.72,P<0.001) or C-MET amplification (χ2=236.41,P<0.001).Conclusions The patients newly diagnosed with lung adenocarcinoma with EGFR mutation plus C-MET amplification demonstrate poor histological differentiation,rapid progress,and poor prognosis.The patients are often in the advanced stage when being diagnosed with cancer.Attention should be paid to this concurrent adverse driving molecular event in clinical work.With increasing availability,the inhibitors targeting C-MET may serve as an option to benefit these patients in the near future.

Analysis of clinicopathological features of seven patients with lung adenocarcinoma translating to small cell lung cancer / 中国肿瘤临床

Objective: To analyze the clinicopathological features of small cell lung cancer transformed from lung adenocarcinoma. Methods: We retrospectively analyzed the clinical and pathological characteristics and follow-up data of seven patients who had been diagnosed with small cell lung cancer transformed from lung adenocarcinoma following treatment from January 2014 to December 2018 at the Fourth Hospital of Hebei Medical University. Results: The latest follow-up had been performed on June 1, 2020. The median time of small cell lung cancer transformation from lung adenocarcinoma following treatment was 31 months; the median time of tyrosine kinase inhibitor (TKI) application before transformation is 14 months. Three patients had transformation at the same site as the original. Seven patients had higher levels of neuron-specific enolase (NSE) before transformation. Before the transformation, disease progression mostly occurred at multiple sites, and the lung, bone, brain, pleura, and lymph nodes were commonly affected. In all cases, immunohistochemical indicators after transformation showed that thyriod transcription factor-1 (TTF-1) was positive; Napsin A was negative; Syn, CD56, and AE1/AE3 were positive; Ki67 expression was high; and PD-L1 expression was negative. Genetic testing after transformation showed that six patients had maintained the original mutant EGFR gene. Treatments after transformation were mainly comprehensive, based on chemotherapy. The median progression-free survival time after transformation was 6 months, and median survival time after transformation for five patients who died was 10 months. Conclusions: Once lung adenocarcinoma undergoes transformation to small cell lung cancer, the disease progresses rapidly, and survival time is short. Patients with lung adenocarcinoma due to EGFR E19 mutation who undergo EGFR-TKI therapy are more prone to small cell lung cancer transformation, and the time to transformation generally exceeds 2 years. The sites of disease progression before transformation are often multiple, and NSE is increased. After transformation, patients generally maintain the original EGFR mutation.

Study of EGFR mutation status in multiple pulmonary ground-glass nodules / 中国肿瘤临床

Objective: To compare the mutation status of epidermal growth factor receptor (EGFR) between different lesions and clini-cal characteristics of synchronous multiple ground-glass nodules (SMGGNs). Methods: A retrospective analysis was conducted using clinical data from 35 patients with SMGGNs who were admitted to and received surgery at The Fourth Hospital of Hebei Medical Uni-versity Hospital from January 2017 to December 2018. Next generation sequencing (NGS) was performed for all surgical specimens to detect the mutation status of exons 18, 19, 20, and 21 of the EGFR gene to analyze the relationship between the EGFR mutation sta-tus of the lesions and patient gender, age, lesion location, imaging manifestation of nodules, and adenocarcinoma pathological type . Results: The EGFR mutation rate was 65.7% (23/35 patients). Non-smoking patients and females had higher EGFR mutation rates (P=0.015, P<0.001). The EGFR mutation rate of invasive adenocarcinoma nodules was higher than those of atypical adenomatous hyper-plasia, adenocarcinoma in situ, and minimally invasive adenocarcinoma ( P<0.001). Exon 19 deletion and L858R mutation were the most common mutations of the EGFR gene. There was no significant difference between the pathological subtypes of adenocarcino-ma and the EGFR mutant subtype (P=0.707). Among the 27 patients with multiple nodules with detectable EGFR mutations, the EGFR mutation rate was 85.2% (23/27 patients). Conclusions: The EGFR gene mutation status is different in patients with multiple pulmo-nary ground-glass nodules, suggesting that the occurrence and development of each nodule are independent events. EGFR gene muta-tion is closely related to the development of ground-glass nodules, especially in the invasion of tumors.

Relationship between driver gene mutation and clinicopathological features in 300 cases of non-small cell lung cancer based on next generation sequencing / 临床与实验病理学杂志

Purpose To explore the mutation characteristics of common driver genes in non-small cell lung cancer (NSCLC) and its relationship with clinicopathological features.Methods Next generation sequencing (NGS) was used to detect the mutations of common driving genes such as EGFR, KRAS, ALK, ROS1, BRAF, MET, RET and HER-2 in 300 paraffin-embedded NSCLC tissues. Results In 300 patients with NSCLC, the mutation rates of EGFR, KRAS, ALK, ROS1, BRAF, MET, RET, HER-2 were 52.00％, 10.33％, 6.67％, 1.67％, 3.67％, 3.33％, 1.00％, and 2.33％, respectively. A case of EGFR 21 exon L858 R mutation was combined with LINCO1446-EGFR gene fusion. EGFR 20 th exon C797 S and T790 M existed in cis or trans form and merged with EGFR sensitive mutations in 1 case each. 3 cases of EGFR gene point mutation was associated with MET gene copy number amplification. EGFR mutations were more commonly detected in non-smoking women with lung adenocarcinoma (P<0.05).KRAS mutations were more commonly found in smoking men (P<0.05). ALK mutations were associated with age (P<0.05), and more commonly noted in patients younger than 60 years.ROS1 fusion mutations were associated with gender (P<0.05), more commonly detected in women. BRAF, MET, RET, and HER-2 gene mutations were not associated with gender, age, smoking, histological type, and c TNM stage. Conclusion EGFR can coexist with other driver gene mutations. Gene mutations and clinicopathological features like gender, age, smoking, and histological types have corresponding links. The incidence of BRAF, MET, RET, and HER-2 mutations is low, and its clinical significance remains to be explored. Coexisting gene mutations and rare mutations discovered by NGS should be taken seriously.

Construction of fused recombinant vector of EGFR gene containing G719S and T790M by overlap PCR / 临床检验杂志

Objective To construct the recombinant pMD19-exon18-exon20 plasmid containing locus G719S and T790M of EGFR gene associated with cervical cancer,which may provide a template for preparing the mutant recombinant vector of EGFR gene.Methods Using the healthy human genome DNA as templates,the segments of exon 18 and exon 20 of EGFR gene were amplified by two pairs of specific primers which were designed based on the sequences of overlapping and complementary area.The amplified segments were linked by overlap PCR.The products of linked exon18-exon20 were further inserted into the vector pMD19-T.The constructed recombi nant pMD19-exon18-exon20 plasmid was finally transformed into competent cells E.coli DH5α and then identified by PCR with bacterial solution and genome sequencing.Results The amplified fragments of exon18 and exon20 were clearly appeared at 778 bp and 596 bp and the fused product of exon18-exon20 was showed at 1 374 bp on agarose gel electrophoresis.The recombinant plasmid of fusion EGFR gene was consistent with the expected results via bacterial PCR assay and DNA sequencing.Conclusion We successfully fused the segments of exon18 with exon20 and constructed the recombinant expression vector of EGFR gene by using overlap PCR method.

Statistical analysis of EGFR mutation and EML4-ALK gene fusion in non-small cell lung cancer patients of Zhongshan City / 国际检验医学杂志

Objective To detect the mutation of epidermal growth factor receptor(EGFR) gene,fusion of echinoderms microtubule associated protein sample-4 and gradual change of lymphoma kinase(EML4-ALK) gene,as well as describe their relationship with the clinicopathological features in patients with non-small cell lung cancer(NSCLC) from Zhongshan city of Guangdong province.Methods Mutations of EGFR gene and EML4-ALK fusion gene in 753 NSCLC patients from Zhongshan People's hospital were detected by ARMS real-time PCR.To study the relationship between the mutation and clinical features and explore the significance of EGFR gene mutation and EML4-ALK fusion in NSCLC.Results The EGFR mutation rate of 753 NSCLC patients is 43.16％(325/753),with highest mutation rate in 19 and 21 exons,43.08％(140/325) and 47.38％ (154/325) respectively,and the main mutation in 21 exon is L858R mutation.EGFR mutation is more common in female/non-smoking patients,or patients with adenocarcinoma/adenosquqmous carcinoma/adenocarcinoma metastasis(P＜0.05),but not relates with the age of patients(P＞0.05).The EML4-ALK fusion gene of 110 patients whose EGFR mutation were checked were simultaneously detected,showing a 9.09 ％ (10/110) mutation rate,and the mutation rate in type 1(80％) is significantly higher than type 2(10％) and 3(10％).Patients with EML4-ALK gene mutation tend to be younger(P＜0.05),but the EML4-ALK gene mutation rates show no significant differences in groups classified by gender,smoking history or pathological classification(P＞0.05).EGFR gene mutation and EML4-ALK fusion were detected in one patient simultaneously.Conclusion The EGFR mutation rate of patients with NSCLC in Zhongshan city is consistent with results reported in domestic and foreign literatures.Detections of EGFR gene mutation and EML4-ALK fusion are necessary test items,providing important evidence in molecular targeting therapy in NSCLS.

Effect of exectinib on the expression of EGFR genes 19 and 21 in patients with advanced lung cancer / 中国生化药物杂志

Objective To investigate the effect of ectectin on the expression of EGFR gene 19 and 21 in patients with advanced lung cancer.Aim to provide scientific reference.Methods68 patients with advanced lung cancer were treated with 3 months of ectectin.Serum free DNA was extracted from patients before and after treatment.RT-PCR was performed to amplify EGFR exon 19 and exon 21, EGFR gene mutations were detected by gene sequencing, and the mutation status of exon 19 and exon 21 of EGFR gene before and after treatment were evaluated.ResultsIn 68 patients with lung cancer before and after treatment with ectatinib, Histopathological types were correlated with the efficacy of ectectin hydrochloride.DCR of adenocarcinoma was significantly higher than that of squamous cell carcinoma.18 cases of EGFR mutations, there is no CR and PD cases, PR 11 cases, SD 7 cases, ORR 61.1%, DCR 100%.In the remaining 50 patients with unknown EGFR status, PR 24, SD 4, PD 15, ORR 48.0%, DCR 56.0%.There was no significant difference in the mutation rate of exon 19 and 21 between before and after chemotherapy.The mutation status of serum EGFR was correlated with the pathological type of patients (P<0.02).The EGFR gene mutation rate was 35.4% (17/48) before chemotherapy and 50% after chemotherapy.The rate of EGFR gene mutation was 55% before and after chemotherapy.ConclusionEctectin is effective in the treatment of advanced lung cancer, and it is effective in lung adenocarcinoma and may lead to the change of serum EGFR gene mutation in lung cancer patients,for the first-line application of patients treated with ectectin hydrochloride should always observe the changes in serum EGFR gene, to avoid blindness in treatment of patients.

Statistical analysis of EGFR mutation and EML4-ALK gene fusion in non-small cell lung cancer patients of Zhongshan City / 国际检验医学杂志

Objective To detect the mutation of epidermal growth factor receptor(EGFR) gene,fusion of echinoderms microtubule associated protein sample-4 and gradual change of lymphoma kinase(EML4-ALK) gene,as well as describe their relationship with the clinicopathological features in patients with non-small cell lung cancer(NSCLC) from Zhongshan city of Guangdong province.Methods Mutations of EGFR gene and EML4-ALK fusion gene in 753 NSCLC patients from Zhongshan People's hospital were detected by ARMS real-time PCR.To study the relationship between the mutation and clinical features and explore the significance of EGFR gene mutation and EML4-ALK fusion in NSCLC.Results The EGFR mutation rate of 753 NSCLC patients is 43.16％(325/753),with highest mutation rate in 19 and 21 exons,43.08％(140/325) and 47.38％ (154/325) respectively,and the main mutation in 21 exon is L858R mutation.EGFR mutation is more common in female/non-smoking patients,or patients with adenocarcinoma/adenosquqmous carcinoma/adenocarcinoma metastasis(P＜0.05),but not relates with the age of patients(P＞0.05).The EML4-ALK fusion gene of 110 patients whose EGFR mutation were checked were simultaneously detected,showing a 9.09 ％ (10/110) mutation rate,and the mutation rate in type 1(80％) is significantly higher than type 2(10％) and 3(10％).Patients with EML4-ALK gene mutation tend to be younger(P＜0.05),but the EML4-ALK gene mutation rates show no significant differences in groups classified by gender,smoking history or pathological classification(P＞0.05).EGFR gene mutation and EML4-ALK fusion were detected in one patient simultaneously.Conclusion The EGFR mutation rate of patients with NSCLC in Zhongshan city is consistent with results reported in domestic and foreign literatures.Detections of EGFR gene mutation and EML4-ALK fusion are necessary test items,providing important evidence in molecular targeting therapy in NSCLS.

Clinical effects of icotinib on lung denocarcinoma patients with unknown EGFR gene status and poor performance status / 重庆医学

Objective This research is aimed to investigate the efficacy and toxicity of icotinib for lung adenocarcinoma pa-tients with poor performance status and unknown EGFR gene status .Methods A total of 27 lung adenocarcinom patients with poor Eastern Cooperative Oncology Group-Performance status(ECOG-PS) and unknown EGFR gene status referred to Chongqing Canc-er Institute from August 2012 to August 2014 were analyzed .Icotinib (125 mg) was orally administered three times per day .Asess the efficacy and adverse reaction ,calculate survival rates .Results Among the 27 patients ,the objective response rate(ORR) and disease control rates(DCR) were 29 .6% and 81 .5% ,respectively .The median progression free survival time was 6 .0 months .A to-tal of 70 .4% of patients had an significant improvment in ECOG-PS scores ,following icotinib treatment (Z= - 2 .157 ,P= 0 .031) . Fatigue ,anorexia and diarrhea were the most frequent adverse reaction ,which defined as grade 1 to 2 rashes .Conclusion Lung ade-nocarcinoma patients with poor performance status and unknown EGFR gene status may benefit from icotinib therapy ,and patients were tolerated well .

Research progress of molecular biology in ground-glass opacity / 中国癌症杂志

With the low-dose spiral computed tomography (LDCT) widely applied in screening for early-stage lung cancers recently, positive rates of Ground-Glass Opacity (GGO) are gradually increased. Many professional researchers believe GGO has a close relationship with the early-stage lung adenocarcinoma, consequently, qualitative diagnosis and early treatment of GGO plays a signiifcant role in improving the diagnostic and survival rates of patients with early-stage lung cancer. Since relative imaging diagnosis, location methods and surgical approaches of GGO have been reported a lot by domestic and overseas researchers, our reviews would mainly focus on the diverse research progress of molecular biology of GGO in the past several years.

Clinical applicability of xTAG liquidchip technology as EGFR mutations detection method in lung cancer / 临床与实验病理学杂志

Purpose To analyze EGFR exon 18 ~21 gene mutations in lung cancer, and compare xTAG liquidchip technology and Sanger sequencing technology in clinical practice. Methods 1 139 tumor tissue samples from phaseⅠtoⅣlung cancer patients were randomly collected. DNA was extracted from the samples. EGFR gene mutation status in exon 18~21 was detected by xTAG liquidchip technology and Sanger sequencing technology respectively. Results The mutation status of EGFR was obtained by xTAG liquidchip technology in 1 134 patients, and 1 105 by Sanger sequencing technology, detection success rate was 99. 56% and 97. 01% respective-ly. The sensitivity and specificity of xTAG liquidchip technology Comparing with Sanger sequencing was 99. 59% and 94. 54%. Sever-al cases of multiple mutations were detected by both methods. All mutation types detected by two methods are fully consistent. Conclu-sions Comparing with Sanger sequencing technology, xTAG liquidchip technology, which is able to detect mutations of exon 18~21 simultaneously, is more convenient and efficient for EGFR gene mutation detection in lung cancer.

Detecting KRAS,EGFR mutations in nonsmall-cell lung cancer/colorectal cancer patients by high resolution melting curve analysis / 中国基层医药

ObjectiveTo evaluate the feasibility of detecting the mutations of KRAS,EGFR in nonsmall-cell lung cancer and colorectal cancer patients with high resolution melting curve analysis. MethodsAt first,the mutations in KRAS exon 2 and EGFR exons 18 to 21 of 5 patients with non-small cell lung cancer and 5 patients with colorectal cancer were detected by high resolution melting curve analysis.Then the results were confirmed with direct sequencing. ResultsBy high resolution melting curve analysis,1 of 10 patients was found to carry EGFR 19 mutation who was harboring 2236-2250del mutation.By direct sequencing,consistent results of the patients with mutations orwithout mutations were got. Conclusion s The new high resolution melting curve analysis was more efficient,more convenient than direct sequencing.Moreover,it was a low-cost test,which was suitable for clinic test.

Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells / 重庆医科大学学报

Objective:To construct a expression vector of EGFR specific siRNA and transfect it into human ovarian cancer cell line SKOV3,then to evaluate the effect of RNA interference(RNAi) on the target gene expression and radiosensitivity of the cell line.Methods:EGFR sh-RNA was synthesized in vitro and transfected into SKOV3 cells with lipofectamine technique and then the positive clones were selected by G418.The positive RNA interference cell line,negative RNA interference cell 1ine and the normal control cell 1ine were included in the study.RT-PCR and Western Blot technique were performed to detect the inhibitory effect of RNAi on EGFR mRNA level and protein level,respectively.Three kinds of cell lines were exposed to different doses of radiation.Cell survival curve was drawn by means of MTT method.Results:The expression of EGFR gene was surpressed obviously in the specific sequence shRNA group,and EGFR mRNA and protein surpression rates were 77.5% and 76.1%,respectively;Sequence-specific shRNA-EGFR can significantly enhance the radiosensitivity of ovarian cancer cells,and significant statistical differences existed among the three groups(P