A Case of Non-small Cell Lung Cancer Treated with Three ALK Inhibitors and Chemotherapy / 中国肺癌杂志

The echinoderm microtubule associated protein-like 4 (EML4) and anaplastic lymphoma kinase (ALK) were fractured and fused to become EML4-ALK. Most of these EML4-ALK-positive non-small cell lung cancer patients respond well to the ALK inhibitor. Many patients can benefit from drug target therapy for a long time, and some patients can achieve long-term survival of more than 7 years under the optimized treatment mode. This patient has lung adenocarcinoma positive for EML4-ALK fusion gene, but the treatment outcome is obviously different from that of other patients with lung cancer positive for EML4-ALK fusion gene. After the first to third generations of ALK inhibitor targeted therapy and chemotherapy, the disease progresses rapidly, the drug resistance time is short, the survival time is short, and the benefit is limited. The patient received targeted therapy of Crizotinib, Ceritinib and Lorlatinib successively from July 15, 2019, followed by two chemotherapy courses of Bevacizumab combined with Pemetrexed and Carboplatin. The patient died on September 10, 2020, with a survival of 15 months. At the same time, the treatment showed common adverse reactions of ALK inhibitors. This paper analyzed the therapeutic effect and treatment dilemma of this patient, and provided an exploration direction for the treatment of patients with EML4-ALK fusion gene positive lung cancer.
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Efficacy and Safety of Crizotinib in Advanced or Recurrent ALK-positive Non-small Cell Lung Cancer / 中国肺癌杂志

BACKGROUND@#Anaplastic lymphoma kinase (ALK) positive in non-small cell lung cancer (NSCLC) was about 5%-7% and ALK tyrosine kinase inhibitor (TKI) was the standard treatment in NSCLC. The aim of this study is to evaluate the efficacy and safety of crizotinib in patients with advanced ALK gene-positive or recurrent NSCLC.@*METHODS@#Three methods were used to screen patients with advanced or recurrent NSCLC harboring ALK gene fusion/translocation. The patients with ALK positive tested by flourescence in situ hybridization (FISH) was given orally crizotinib, 250 mg, bid. The objective response rate (ORR), progression-free survival (PFS) and safety were evaluated.@*RESULTS@#A total of 226 patients were screened, 39 of whom had ALK fusion or translocation, and 37 were enrolled in the study. 35 patients were evaluated for objective response, ORR was 70.3%, and disease control rate (DCR) was 94.6%, and median PFS was 11.8 mon. The main adverse reactions were elevated transaminase (Grade 1, 91.7%), elevated transaminases (Grade 2, 23.4%), nausea (Grade 1, 75.6%), anemia (Grade 1-2, 62.3%), visual impairment (Grade 1, 21.8%), weight loss (Grade 1, 31.4%), pneumonia (Grade 2, 3.5%).@*CONCLUSIONS@#Crizotinib can be used for the treatment of advanced NSCLC with ALK fusion/translocation. It is highly effective and well tolerated.

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.

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.

Distribution and clinical significance of EML4-ALK fusion gene in phase Ⅰ lung cancer / 中华胸心血管外科杂志

Objective To detect the mutation frequency of EML4-ALK fusion gene in lung cancer patients, and to inves-tigate the distribution of mutation character for EML4-ALK fusion gene in Ⅰ stage lung cancer patients and clinical features as well as provide a reference for the individual treatment of lung cancer .Methods 256 fresh tumor tissue specimens of lung cancer patients were screened from the specimen bank of our hospital and all the patients had accepted the surgical treatment from February 2013 to December 2014.Total RNA was extracted and then be transcribed into cDNA, the amplification-refrac-tory mutation system(ARMS) was used to detect mutation of EML4-ALK fusion gene.The results according to the positive con-trol, negative control and RNA quality control for EML4-ALK fusion type were analyzed.Results During the 256 patients ofⅠ stage lung cancer, there were 17 patients(6.64%) had mutations in EML4-ALK fusion gene.In lung adenocarcinoma mu-tation rate(16/207, 7.73%) was higher than that of lung squamous cell mutation rate(1/39, 2.56%), lung adeno-squamous mutation rate(0/4, 0) and large cell carcinoma(0/5, 0) of the mutation rate;young lung cancer patients( <63 years) of the mutation rate(14/139, 10.07%) was significantly higher than the high age of lung cancer patients(≥63 years old) mutation rate(3/117, 2.56%), P =0.009.EML4-ALK fusion with tumor invasion and visceral pleura group incidence (9/80, 11. 25%) was significantly higher than that of non-invasive and visceral pleura group incidence rate(8/176, 4.55%), P =0.045.Conclusion The occurence of EML4-ALK fusion correlates with patients’ age as well as whether visceral pleura is in-vaded, type 1 EML4-ALK fusion was detected more in phase I lung cancer patients.

Clinical value of antibody D5F3 and Survivin in non small cell lung cancer / 中国生化药物杂志

Objective To investigate the clinical value of antibody D5F3 and Survivin in patients with non small cell lung cancer.Methods 200 paraffin embedded specimens of patients with non small cell lung cancer tested by RT-PCR(including EML4-ALK mutant and wild type)from October 2012 to June 2014 were selected.The gene protein expression were tested by ALK new antibody D5F3 and the sensitivity and specificity were compared by negative and positive(+～3+).ResultsTest results show(+)the coincidence rate is 15.78％,(+ +)the coincidence rate is 27.27％and(+ + +)compliance rate was 87.5％,the difference was statistically significant(P<0.05).The expression of Survivin protein in NSCLC tissues was correlated with clinicopathological features.The positive rate of Survivin protein expression was correlated with the degree of differentiation(P<0.05),but not with other clinical and pathological features,with the decrease of differentiation,the positive rate of Survivin protein expression was significantly increased.Conclusion D5F3 and Survivin antibodies are highly sensitive and specific in patients with NSCLC,with the screening value,save social resources,for the majority of patients with lung cancer services.

Clinical value of Antibody D5F3 and Survivin in patients with non- small cell lung cancer / 中国生化药物杂志

Objective To investigate the clinical value of D5F3 and Survivin in patients with non-small cell lung cancer. Methods 100 patients with non - small cell lung cancer admitted in our hospital from January 2016 to June 2017 were examined by immunohistochemical method, to analyze the clinical value of D5F3 and Survivin antibody in non - small cell lung cancer (NSCLC). Results The coincidence rate of D5F3 was (20％), (++) was 33.33％, and the coincidence rate was (87.50％), and the difference was statistically significant (P<0.05). The positive rate of Survivin protein expression was correlated with the differentiation degree of non-small cell lung cancer, TNM stage and lymph node metastasis (P<0.05). Conclusion D5F3 has high accuracy, sensitivity and specificity, and has important clinical diagnostic value; The detection of Survivin expression is helpful for clinical differentiation, clinical staging and lymph node metastasis in patients with non-small cell lung cancer.

Bypass signaling pathway activation mediates resistance of EML4-ALK fusion gene positive lung cancer cell line H3122 to alectinib / 中国病理生理杂志

AIM:To detect the changes of active status of bypass signaling pathways in EML4-ALK positive lung cancer cell line H3122 treated with alectinib, hepatocyte growth factor (HGF), epidermal growth factor (EGF) and transforming growth factor-α (TGF-α), and to explore the potential mechanisms.METHODS:EML4-ALK positive cell line H3122 was treated with increasing concentrations of alectinib or/and induced by HGF, EGF and TGF-α.The cell viability was measured by CCK-8 assay.The cell apoptosis was analyzed by flow cytometry.The protein levels and phosphorylation status of ALK, c-Met and EGFR, and the downstream molecules AKT, ERK, p-AKT and p-ERK were examined by Western blot.RESULTS:The viability of the H3122 cells was inhibited by alectinib in a dose-dependent manner after administrated for 72 h, and the IC50 value was 0.042 μmol/L.The concentration-growth curves of the H3122 cells shifted to the right after induced by HGF, EGF and TGF-α.After treatment with alectinib at 0.05 μmol/L for 48 h, the apoptotic rate of H3122 cells was (20.12±1.36)%, while the apoptotic rates of the cells in the groups of alectinib combined with HGF, EGF or TGF-α were (7.85±1.03)%, (5.60±0.79)% and (4.58±1.00)%, respectively.Those values were remarkably lower than those in alectinib single treatment group (P<0.05).Alectinib inhibited the protein levels of p-ALK and its downstream signaling pathway molecules, while HGF significantly up-regulated the protein levels of p-Met and its downstream p-AKT and p-ERK.Besides, EGF and TGF-α remarkablely up-regulated the protein levels of p-EGFR and its downstream p-AKT and p-ERK.Combined treatment with crizotinib and 17-DMAG successfully inhibited the viability of the H3122 cells even in the presence of the HGF and EGFR ligands, respectively.CONCLUSION:Bypass signaling pathways are activated by HGF, EGF and TGF-α in EML4-ALK positive lung cancer cell line H3122, which may be linked to alectinib resistance.

EML4-ALK and EGFR mutation status and survival analysis in Uygur with stageⅣNSCLC / 天津医药

Objective To investigate the relationship between the echinoderm microtubule associated protein like 4-anaplastic lymphoma kinase (EML4-ALK) and epithelial growth factor receptor (EGFR) mutation status and overall survival (OS) in Uygur patients with stageⅣnon-small cell lung cancer (NSCLC) who did not accept tyrosine kinase inhibitor treat-ment. Methods Totally 97 tissue samples were collected from Uygur patients with stageⅣNSCLC who did not accept tyro-sine kinase inhibitor treatment. EML4-ALK fusion gene and EGFR mutation status were detected by using FISH and ARMS methods. The survival rates were analysed. Results In 97 tissue samples, EML4-ALK fusion genes were found in 6 (6.2%) samples, EGFR mutations were found in 26 (26.8%) samples. The survival analysis showed that there was no significant dif-ference in OS between EML4-ALK fusion gene group and no EML4-ALK fusion gene group (P=0.941). There was no signifi-cant difference in OS between EGFR mutation group and wild-type EGFR group (P=0.607). The values of median OS were 17.7 months, 17.3 months and 16.2 months for EGFR mutant group, EML4-ALK positive group and EML4-ALK negative+EGFR wild-type group, and thre was no significant difference between them (P=0.915). Conclusion Excluding the thera-peutic influence in TKIs, EML4-ALK fusion gene and EGFR mutation status of tumor tissue can not be used as an indepen-dent factor in assessing the prognosis in Uygur patients with stageⅣNSCLC.

Research progress of Alectinib in the treatment of non-small cell lung cancer / 实用肿瘤学杂志

Recently,targeted therapy plays an important role in the treatment of non-small cell lung cancer.Crizotinib,the first generation of anaplastic lymphoma kinase( ALK) tyrosine kinase inhibitor,has been ap-proved for the treatment of ALK-rearranged NSCLC in the US since 2011.However,the crizotinib therapy is ef-fective only in the early stage of treatment.After a long time treatment,therapy resistance will follow because of the emerging second mutation of the tumor cell.In recent years,as the second generation of anaplastic lymphoma kinase tyrosine kinase inhibitor, alectinib has been approved in Japan for the treatment of ALK -rearranged NSCLC patients.Thus,research progress of Alectinib for NSCLC therapy is summarized in this article.

Alectinib: a novel second generation anaplastic lymphoma kinase (ALK) inhibitor for overcoming clinically-acquired resistance

The development of inhibitors for the tyrosine anaplastic lymphoma kinase (ALK) has advanced rapidly, driven by biology and medicinal chemistry. The first generation ALK inhibitor crizotinib was granted US FDA approval with only four years of preclinical and clinical testing. Although this drug offers significant clinical benefit to the ALK-positive patients, resistance has been developed through a variety of mechanisms. In addition to ceritinib, alectinib is another second-generation ALK inhibitor launched in 2014 in Japan. This drug has a unique chemical structure bearing a 5H-benzo[b]carbazol-11(6H)-one structural scaffold with an IC50 value of 1.9 nmol/L, and is highly potent against ALK bearing the gatekeeper mutation L1196M with an IC50 of 1.56 nmol/L. In the clinic, alectinib is highly efficacious in treatment of ALK-positive non-small cell lung cancer (NSCLC), and retains potency to combat crizotinib-resistant ALK mutations L1196M, F1174L, R1275Q and C1156Y.

EML4-ALK fusion gene and non-small cell lung cancer / 第二军医大学学报

The fusion gene of echinoderm microtubule-associated protein-like 4 (EML4) and anaplastic lymphoma kinase(ALK) has recently been identified as a new molecular target for non-small cell lung cancer (NSCLC). EML4-ALK fusion gene is about 3%-11% in patients with NSCLC, and it has a higher morbidity in NSCLC patients who are young, without or with light smoking history and with adenocarcinomas. EML4-ALK positive NSCLC patients may be treated with ALK inhibitor (crizotinib). This review focuses on the biology, detection method, clinical characteristics and the therapeutic application of EML4-ALK in NSCLC.

Advances in assay of EML4-ALK gene rearrangements in non-small-cell lung cancer / 军事医学

Echinoderm microtubule associated protein like 4-anaplastic lymphomakinase ( EML4-ALK) rearrangement is another driver-mutation in non-small-cell lung cancer (NSCLC) besides epidermal growth factor receptor (EGFR).Accu-rate identification of EML 4-ALK lung adenocarcinoma is essential for the selection of appropriate therapy .Immunohisto-chemistry (IHC), fluorescence in situ hybridization (FISH), and polymerase chain reaction (PCR) are the main three assay methods for clinical practice .Recently, researchers have done lots of work on the exploration of detection , diagnosis, and treatment prediction of carcinoma based on serum proteomics .In this paper, recent developments in these fields are reviewed.

EML4-ALK rearrangement in cancer tissue and serum of non-small cell lung cancer patients / 国际肿瘤学杂志

Objective To observe EML4-ALK fusion gene mutation expression rate in serum and cancer tissue of patients with non-small cell lung cancer (NSCLC) in Chinese populations,and the consistency of mutation in serum and cancer tissues,and the feasibility of real-time,dynamic detection of EML4-ALK fusion gene therapy by using FQ-PCR.Methods 123 cases of serum and 98 cases of tissue of NSCLC patients were detected by fluorescence quantitative polymerase chain reaction,and 77 cases of which were matched.The clinical curative effects of ALK inhibitor (crizotinib) were analyzed.Results 13 rearrangement in 123 (10.6％) of the patients'serum samples and 11 rearrangement in 98 (11.2％) tumor tissue.EML4-ALK rearrangement were mainly discovered in adenocarcinoma (x2 =4.083,P =0.036),and non-smokers in NSCLC (x2 =5.326,P =0.019).The consistency of patients with EML4-ALK matched tumor tissue and serum reached 66.7％ (6/9,κ =0.779).The EML4-ALK fusion gene rearrangement in patients receiving ALK inhibitor (crizotinib) treatment achieved significant benefit.Conclusion The EML4-ALK rearrangement mainly exists in the serum and tumor tissue of adenocarcinoma and non-smokers in NSCLC.When tumor tissue samples are unable to be obtained,FQ-PCR can be used to detect serum EML4-ALK fusion gene mutation for selecting NSCLC patients suitable for crizotinib therapy instead of tumor tissue.

Successful Treatment of Crizotinib-Associated Severe Hepatotoxicity in Anaplastic Lymphoma Kinase-Positive Non-Small Cell Lung Cancer

Crizotinib-associated severe hepatotoxicity has been rarely reported and experts recommended stopping crizotinib treatment in patients with grade 3/4 transaminase elevation. We experienced a case of anaplastic lymphoma kinase-positive non-small cell lung cancer occurring as a result of severe hepatotoxicity due to crizotinib-associated hepatitis, accompanied by the reactivation of chronic hepatitis B, which was reversed with dose reduction and anti-viral therapy. Our case highlights the possibility that crizotinib might induce hepatitis and this might be associated with the underlying presence of chronic hepatitis B. In addition, crizotinib could be continued with reduced unless there are any other therapeutic options.

Role of mTOR signaling pathway in crizotinib-induced apoptosis of EML4-ALK fusion gene-positive lung adenocarcinoma cell line H2228 / 中国病理生理杂志

AIM:To investigate the mammalian target of rapamycin ( mTOR) signaling pathway as the center playing a role in the crizotinib-induced apoptosis of non-small cell lung cancer (NSCLC) cell line H2228, which represents positive echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene. METHODS:H2228 cells were processed according to different purposes .Fluorescence quantitative PCR is used to ob-serve the gene states .MTT assay is used to detect the cell inhibition rates .The cell apoptosis and cell cycle were analyzed by flow cytometry .The expression and activation levels of the key proteins in the mTOR signaling pathway were determined by Western blotting .RESULTS:Crizotinib promoted the apoptosis of H 2228 cells in a time-and dose-dependent manner . Crizotinib blocked the H2228 cells staying at the G1 phase.In apoptotic H2228 cells processed with crizotinib, the activa-tion level of mTOR was decreased , and the activation levels of the key proteins in upstream and downstream of mTOR path -way were both declined .The expression level of the fusion protein EML 4-ALK variant 3 was not affected , but its active form of p-ALK was significantly suppressed .CONCLUSION:mTOR signaling pathway has a certain relationship with the crizotinib-induced apoptosis of lung cancer cell H 2228, which represents positive EML4-ALK fusion gene.

Biological characteristics and therapeutic application of EML4-ALK fusion gene in non-small cell lung cancer / 肿瘤

The fusion gene between echinoderm EML 4 (microtubule-associated protein-like 4) and ALK (anaplastic lymphoma kinase) has been identified in non-small cell lung cancer (NSCLC). EML4-ALK is most commonly detected in never smokers with NSCLC and has unique pathologic features. EML4- ALK is oncogenic both in vitro and in vitro . ALK inhibitor (crizotinib) has demonstrated a remarkable clinical efficacy in EML4-ALK-positive NSCLC patients. This review emphasizes the biological and clinical characteristics and the therapeutic application of EML4-ALK in NSCLC. © 2012 by Tumor.

Progress of EML4-ALK fusion gene non-small cell lung cancer inhibitor / 国际肿瘤学杂志

EML4-ALK fusion gene is a new molecular type of non-small cell lung cancer,which is formed by inversion of two gene segments on chromosome 2.Up to now,9 subtypes have been found and all of them have deteriorated and carcinogenic abilities. The detection rate of this fusion gene is much higher in patients who have adenocarcinoma,never or seldom smoke and lack EGFR/KRAS mutation.This fusion gene codes ALK tyrosine kinase,that abnormally phosphorylates downstream products through signaling pathways,leading to canceration.Therefore,the specific inhibitors which target this fusion gene could inhibit its expression and have good treatment effect.At present,the relevent inhibitors which are undergoing clinical trials are PF02341066 and TAE684.

Clinicopathologic and Molecular Characteristics of Lung Adenocarcinoma Arising in Young Patients

Lung cancer rarely occurs in young patients. Recent studies have demonstrated that epidemiologic data are closely correlated to some molecular characteristics. We investigated the clinicopathologic characteristics of lung adenocarcinoma in young patients and evaluated immunohistochemically detected epidermal growth factor receptor (EGFR) mutation status and anaplastic lymphoma kinase (ALK) positivity. Among lung adenocarcinoma patients, 31 cases were of the 50 yr-old group. Young patients were more likely to be females (67.7% vs 40.2%), and nonsmokers (58.1% vs 45.2%) and more often had high TNM stage (stage IV was 80.6% vs 52.1%) and had a high rate of distant metastasis (51.6% vs 28.0%) compared with older patients. The signet ring cell feature was more common (25.8% vs 11.5%) and lepidic growth pattern was rarely present (3.2% vs 16.5%) in the adenocarcinoma of young patients. There was no significant survival difference between the two age groups. The rate of EGFR mutation status and ALK positivity did not show a statistical difference between two groups. In conclusion, lung adenocarcinoma of young patients demonstrates distinct pathologic features with frequent presence of a signet ring cell feature and rare occurrence of lepidic growth pattern. Further investigation for other genetic abnormalities would be needed.

EML4-ALK fusion oncogene and non-small cell lung cancer / 中国癌症杂志

Non-small cell lung cancer(NSCLC) is one of the most life-threatening human malignancies due to its high morbidity and mortality rates.Molecular targeted therapy is promising in treating some patients with NSCLC.However,it is still challenging to select proper candidates.The EML4-ALK fusion oncogene represents a novel molecular target which appears mainly in lung adenocarcinoma,and appears to be a potential biomarker associated with resistance to EGFR tyrosine kinase inhibitors.This review was intended to outline current status of preclinical and clinical research of this molecule.