Related Experiment Video
Updated: Jun 12, 2025
Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure
Published on: August 11, 2017
EGFR T790M mutation detection in NSCLC patients resistant to tyrosine kinase inhibitor therapy
Rabiga Kadyrbayeva1,2, Dilyara Kaidarova3,4, Oxana Shatkovskaya4
1Chemotherapy Center, Kazakh Institute of Oncology and Radiology, Almaty, Republic of Kazakhstan - kadyrbayevarabiga48@hotmail.com.
Background:
The finding of mutations that activate epidermal growth factor receptor (EGFR) in people with lung adenocarcinoma resulted in the creation of a new class of biological treatments called tyrosine kinase inhibitors (TKI). These medications have changed how patients with EGFR mutations are clinically managed, nearly doubling their survival rate compared to standard chemotherapy. Though 1st and 2nd generation EGFR TKIs are initially highly effective, typically within 9-14 months all tumors with the mutation progress due to secondary resistance mutations involving alternative molecular pathways. In most cases (up to 60%), this is due to the T790M mutation emerging in the EGFR gene.
Methods:
The study included 85 patients with NSCLC with progression of the disease after treatment with TKI 1st and 2nd generation. The T790M mutation was determined by digital polymerase chain reaction (PCR) on the QIAcuity One 5plex digital PCR system and traditional real-time PCR. Real-time PCR analysis of the presence of the T790M mutation was performed using the Therascreen EGFR Plasma RGQ PCR Kit (Qiagen). Using a digital PCR system in QIAcuity One (Qiagen) nanoplanets, the T790M mutation was analysed by digital PCR. The age of the patients ranged from 37 to 85 years.
Results:
Of 85 patients with NSCLC with disease progression after TKI treatment, T790M mutations were detected during digital PCR in 30 of 85 patients, which is 35.2% of the sample, and with traditional real-time PCR, positive mutations came out only in 3 out of 85 patients.
Conclusions:
Thus, completed study can assert that digital PCR is able to replace traditional real-time PCR as a more preferable method of high-performance quantitative determination of target nucleic acids and has a relatively high sensitivity without compromising high specificity. Results of this research also show that a liquid biopsy using digital PCR provides an opportunity to avoid repeated tissue biopsy in patients who cannot provide a tumor tissue sample suitable for molecular analysis.
Insights
Digital PCR detected the T790M resistance mutation in 35.2% of non-small cell lung cancer patients, significantly outperforming traditional real-time PCR. This method offers a sensitive, specific liquid biopsy alternative, avoiding repeat tissue sampling.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Epidermal growth factor receptor (EGFR) mutations drive lung adenocarcinoma, leading to targeted therapies like tyrosine kinase inhibitors (TKIs).
- While effective, TKIs face acquired resistance, often mediated by the T790M mutation, necessitating advanced detection methods.
Purpose of the Study:
- To compare the efficacy of digital PCR versus traditional real-time PCR in detecting the T790M resistance mutation in non-small cell lung cancer (NSCLC) patients.
- To evaluate digital PCR as a sensitive and specific tool for liquid biopsy in NSCLC management.
Main Methods:
- The study analyzed 85 NSCLC patients with disease progression after first- and second-generation TKI treatment.
- T790M mutation detection was performed using both digital PCR (QIAcuity system) and traditional real-time PCR (Therascreen EGFR Plasma RGQ PCR Kit).
Main Results:
- Digital PCR identified T790M mutations in 30 out of 85 patients (35.2%).
- Traditional real-time PCR detected the mutation in only 3 out of 85 patients, highlighting a significant difference in sensitivity.
Conclusions:
- Digital PCR demonstrates superior sensitivity and specificity for detecting T790M mutations compared to traditional real-time PCR.
- Liquid biopsy utilizing digital PCR offers a viable alternative to tissue biopsy, particularly for patients unable to provide suitable tumor samples.
More Related Videos
09:38Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib
Published on: June 26, 2019
07:59Author Spotlight: Advancements in Molecular Biomarker Testing for Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
