Balkan Journal of Medical Genetics

EPIDERMAL GROWTH FACTOR RECEPTOR MUTATION STATUS AND THE IMPACT ON CLINICAL OUTCOMES IN PATIENTS WITH NON-SMALL CELL LUNG CANCER
Huang HM1, Wei Y1, Wang JJ1, Ran FY1, Wen Y2, Chen QH3, Zhang BF1,*
*Corresponding Author: Dr. Bingfei Zhang, Sinopharm Dongfeng General Hospital, Hubei University of Medicine, No. 16 Daling Road, 442008, Shiyan, Hubei, China. Tel.: + 86-29-8272597, Email: dfzyysszx@163.com
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MATERIALS AND METHODS

Patients A total of 238 NSCLC patients are included in this study and Figure 1 shows the trial profile. Detailed clinical information of these patients was available on the Sinopharm Dongfeng Hospital medical record system. The gender ratio of these patients was 54.6% (n=130) males and 45.4% (n=108) females. 44.5% (n=106) of the 238 patients were aged over 65 years. Of the 227 patients with a known smoking history, 132 patients (58.1%) had never smoked (smoked <100 lifetime cigarettes), 37 patients (16.3%) were former smokers (≥1 year since quitting smoking), and 58 patients (25.6%) were current smokers (still smoking, or <1 year since quitting smoking). There were 211 NSCLC cases (88.7%) with adenocarcinoma, and other histology types were 27 cases (four adenosquamous carcinoma, five large cell carcinoma, and eighteen squamous cell carcinoma). 157 patients (66%) were diagnosed in advanced NSCLC (Stage IIIB-IV) (Table 1) . Tumor specimens Tumor samples were obtained from 238 NSCLC patients in the daily clinical practice between October 2016 and December 2019 at Dongfeng Hospital. Tumor samples were fixed with 10% formalin, embedded in paraffin, and then 5μm thickness sections were cut. Tumor specimens were evaluated to confirm the NSCLC histology by experienced pathologists. All patients provided written informed consent before EGFR mutation testing. All procedures were supervised and approved by Sinopharm Dongfeng General Hospital Ethics Committee (Approval Number: LW-2021-21). EGFR mutation analysis Genomic DNA was extracted using the TIANamp FFPE DNA Kit, according to the manufacturer’s protocol, and DNA quality and purity was measured by Eppendorf Bio Photometer D30. We used commercially available EGFR kits to detect EGFR mutations in exons 18-21 via ARMS-PCR technology. The EGFR kit is able to detect 21 somatic mutation types, namely, 3 point mutations in exon 18 (G719A, G719C, and G719S , which are referred to as G719X); 11 deletions in exon 19 (which are referred to as 19-Del); 2 point mutations (S768I, T790M) and 3 insertion mutations (H773_V774insH; D770_N771insG; V769_D770insASV, which are referred to as 20-Ins) in exon 20; 2 point mutations in exon 21 (L858R and L861Q) (Table 2). The thermocycling conditions were used as following: 1 cycle of 95˚C for 3 min; 45 cycles of 94˚C for 15 sec and 60˚C for 35 sec. The results were analyzed according to the manufacturer’s guideline. Statistical analysis GraphPad Prism 7.0 and SPSS Statistics 22.0 were the software used for statistical analysis. The associations between EGFR mutation status and clinical characteristics, such as gender, age, smoking history were evaluated by Pearson’s χ2 test or the Fisher exact test. PFS and OS were analyzed by the Kaplan-Meier method, and the differences were calculated by a log-rank test. Variables with a p value less than 0.05 in univariate analysis were entered into a multivariate logistic regression analysis to analyze the fa-vorable factors of EGFR mutations. p<0.05 was considered statistically significant.

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