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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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.

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.

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.

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.