An Alternative Method for Screening EGFR Mutation Using RFLP in Non-small Cell Lung Cancer Patients

Kawada, Ichiro; Soejima, Kenzo; Watanabe, Hideo; Nakachi, Ichiro; Yasuda, Hiroyuki; Naoki, Katsuhiko; Kawamura, Masafumi; Eguchi, Keisuke; Kobayashi, Koichi; Ishizaka, Akitoshi

doi:10.1097/jto.0b013e318186fadd

Cited by 44 publications

(34 citation statements)

References 23 publications

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“…This finding may be explained by the fact that thymidylate synthase, one of the main targets of pemetrexed, was found to be differentially expressed among the histotypes of lung cancer, with a lower expression in AC and a higher expression in SCC and small-cell lung cancer (4,5). Although the most important prediction factor of an epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) has been reported to be the EGFR mutation status rather than the histology, the histology of AC remains an important clinical predicting factor for EGFR-TKI, particularly gefitinib, in the case that the mutation status is not obtained (6,7). Bevacizumab, a targeted therapy agent that inhibits tumor angiogenesis, has only been registered for the treatment of non-SCC in the USA, Europe and other countries due to serious hemorrhagic complications in SCC (3,8).…”

Section: Introductionmentioning

confidence: 99%

Identification of microRNAs differentially expressed between lung squamous cell carcinoma and lung adenocarcinoma

Hamamoto

Soejima

Yoda

et al. 2013

Molecular Medicine Reports

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Abstract. Recent advances in the treatment of non-small cell lung cancer (NSCLC) with new agents require accurate histological subtyping at diagnosis to avoid the higher risk of an adverse response and to obtain the maximum therapeutic response. However, interobserver variability, tumor hetero geneity and the degree of differentiation may affect the decision concerning a pathological diagnosis of NSCLC. Therefore, the aim of this study was to identify specific microRNAs (miRNAs) as standardized biomarkers with high sensitivity and specificity in order to distinguish between squamous cell carcinoma (SCC) and adenocarcinoma (AC). Quantitative polymerase chain reaction (qPCR)-based miRNA array analysis was performed to identify microRNAs differentially expressed between SCC and AC using 86 resected NSCLC samples in addition to adjacent normal tissues. The results were confirmed by independent qRT-PCR assays with the same test samples and 88 additional validation samples, and from this we evaluated the usefulness of the identified miRNAs as biomarkers to distinguish between SCC and AC. Three miRNAs (hsa-miR-196b, hsa-miR-205 and hsa-miR-375) were identified. Discriminant analysis combining the three miRNAs appeared to distinguish SCC from AC accurately in the test and validation samples, demonstrating a sensitivity and specificity of 76 and 80%, and 85 and 83%, respectively. hsa-miR-196b, hsa-miR-205 and hsa-miR-375 were identified as biomarkers capable of distinguishing between lung SCC and lung AC. These newly identified miRNAs may prove to be highly valuable molecular markers for the classification of NSCLC histological subtypes and may contribute to the pathogenesis of each subtype of NSCLC.

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Section: Introductionmentioning

confidence: 99%

Identification of microRNAs differentially expressed between lung squamous cell carcinoma and lung adenocarcinoma

Hamamoto

Soejima

Yoda

et al. 2013

Molecular Medicine Reports

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“…Response rates exceeding 70% for patients with EGFR mutations have been reported in several prospective clinical trials (6,7). About 90% of such somatic mutations are clustered in exon 19 (deletion mutation) and exon 21 (point mutation at codon 858) and were found to involve ligand-independent activation and prolonged receptor kinase activity after ligand stimulation (8,9). However, lung adenocarcinomas with these drug-sensitive EGFR mutations that initially respond to EGFR-TKI eventually develop acquired resistance (10)(11)(12).…”

Section: Introductionmentioning

confidence: 99%

The Combination of Multiple Receptor Tyrosine Kinase Inhibitor and Mammalian Target of Rapamycin Inhibitor Overcomes Erlotinib Resistance in Lung Cancer Cell Lines through c-Met Inhibition

Nakachi

Naoki

Soejima

et al. 2010

Molecular Cancer Research

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Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI) show antitumor activity in a subset of non-small cell lung cancer (NSCLC) patients. However, the initial tumor response is followed by recurrence. Several studies have suggested the importance of other receptor tyrosine kinases (RTK) and downstream kinases as potential targets in the treatment of NSCLC. We used the multiple-RTK inhibitor AEE788, which inhibits EGFR, vascular endothelial growth factor receptor, and human epidermal growth factor receptor 2, with and without the downstream kinase inhibitor RAD001 (an inhibitor of mammalian target of rapamycin). AEE788 inhibited cell growth more effectively than did erlotinib in three NSCLC cell lines examined (A549, H1650, and H1975). However, in the EGFR-TKI-resistant cell line H1975 harboring T790M resistance mutation, cell growth inhibition by AEE788 was only mild, and the phosphorylation of its leading targets such as EGFR and vascular endothelial growth factor receptor 2 was not inhibited. In H1975, AEE788 induced significantly greater cell growth inhibition when combined with RAD001 than when used alone. This cooperative effect was not seen with the combination of erlotinib and RAD001. We found that c-Met was highly phosphorylated in this cell line, and the phosphorylated c-Met was inhibited effectively by AEE788. Using a phospho-RTK array, the phosphorylation of c-Met and insulin-like growth factor-I receptor was inhibited by AEE788. These results suggest that upstream RTK inhibitor overcomes the acquired resistance to EGFR-TKI when combined with downstream kinase inhibitor. Thus, the combined inhibition of upstream and downstream RTKs is a promising strategy for the treatment of NSCLC.

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“…The fundamental approach to EGFR mutation detection by sequencing is gradually replaced by other approaches [11][12][13][14]. Many promising PCR-based techniques have been demonstrated to exceed the mutation detection sensitivity of DNA sequencing.…”

Section: Introductionmentioning

confidence: 99%

A novel high‐resolution chipCE assay for rapid detection of EGFR gene mutations and amplifications in lung cancer therapy by a combination of fragment analysis, denaturing CE and MLPA

Minárik¹,

Gassman

Belšánová³

et al. 2010

Electrophoresis

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There is a growing interest in evaluating molecular markers as predictors of response to new generation of targeted cancer therapies. One of such areas is biological therapy targeting epidermal growth factor receptor gene (EGFR) in lung cancer. The testing of tumor tissue is focused on specific EGFR mutations and EGFR gene amplification, since tumors exhibiting positivity of either of the two marker types are highly sensitive towards the treatment. Although traditional methods of DNA sequencing and fluorescence in situ hybridization are still in use for the detection of EGFR mutations and gene amplification, respectively, there is a need for new dedicated techniques with the primary emphasis on simplicity, sensitivity, speed and cost effectiveness. The main purpose of this work was to integrate diverse assays for both EGFR tests onto a single platform to eliminate the need for different instruments and separate processing. We demonstrate a chip capillary electrophoresis (chipCE) application for EGFR mutation detection by a combination of fragment analysis and denaturing CE along with multiplex ligation-dependent probe amplification (MLPA) for evaluation of EGFR amplification. All separations are carried out in denaturing sieving polymer on a modified Bioanalyzer 2100 chipCE instrument running at temperatures of up to 65°C. The main strength of the resulting high-resolution chipCE application is in its simplicity, speed of analysis and minimal amount of sample required for complete testing of EGFR status. Such an approach could potentially fit medium throughput laboratories providing molecular pathology services for clinical oncologists with fast turnaround times and limited consumption of tissue material.

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An Alternative Method for Screening EGFR Mutation Using RFLP in Non-small Cell Lung Cancer Patients

Abstract: RFLP is a useful method for screening EGFR mutations and can also be applied to predicting the sensitivity of NSCLC patients to EGFR-tyrosine kinase inhibitors.

Cited by 44 publications

References 23 publications

Identification of microRNAs differentially expressed between lung squamous cell carcinoma and lung adenocarcinoma

Identification of microRNAs differentially expressed between lung squamous cell carcinoma and lung adenocarcinoma

The Combination of Multiple Receptor Tyrosine Kinase Inhibitor and Mammalian Target of Rapamycin Inhibitor Overcomes Erlotinib Resistance in Lung Cancer Cell Lines through c-Met Inhibition

A novel high‐resolution chipCE assay for rapid detection of EGFR gene mutations and amplifications in lung cancer therapy by a combination of fragment analysis, denaturing CE and MLPA

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