Journal List > Kosin Med J > v.30(2) > 1057046

Kim: Analysis of an EGFR mutation by PNA clamping method in lung carcinoid tumors

Abstract

Background

Pulmonary carcinoid tumors consisting of typical carcinoid tumors (TC) and atypical carcinoid tumors (AC) are rare, accounting for 2–5% of all lung tumors. TC is considered a low-grade tumor with a rate of distant metastasis up to 12%. In contrast, ACs are more aggressive tumors, displaying a metastatic rate up to 70%. Surgery is the treatment of choice; however, the current treatment outcomes of metastatic lung carcinoids are discouraging. This study aimed to investigate the EGFR mutation using the PNA-mediated clamping method and to provide basic data for using EGFR-TK1 and its clinical implications.

Materials and Methods

A total of 14 cases that underwent surgery were diagnosed as carcinoid tumors and pathologically classified as TC and AC. The paraffin-embedded tissues were analyzed for EGFR mutations using the PNA-mediated PCR clamping technique. The mutant type was noted in the cases with aCt greater than 2.0.

Result

Of 14 cases, eight were AC and six cases were TC. No known EGFR mutation was detected with aCt less than 2.0.

Conclusion

The EGFR genotype determined using the PNA-mediated PCR clamping method was wild-type in all pulmonary carcinoid tumors. Therefore, the application of EGFR-TK1 is limited in pulmonary carcinoid tumors.

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Fig. 1.
The four steps of the PCR cycle in PNA ClampTM
kmj-30-141f1.tif
Fig. 2.
The wild type was detected in PNA-mediated clamping real-time PCR for EGFR.
kmj-30-141f2.tif
Table 1.
The clinical and pathological characteristics of 14 cases of pulmonary carcinoid tumors
Case Dx Age Gender Tumor Size (cm) No of mitosis events (/10HPF) Necrosis Lymph node metastasis Neuroendocrine markers
1 T 63 F 2.8 0 +
2 T 41 F 3 1 +
3 T 48 M 5 0 +
4 T 53 M 3.5 1 +
5 T 31 M 1.8 0 +
6 T 59 M 1.2 0 +/-
7 A 34 M 4 6 +
8 A 55 M 5 7 + +
9 A 47 M 4.8 5 +
10 A 61 M 4.5 10 + +
11 A 54 M 3.3 4 + +
12 A 72 M 5 4 + +
13 A 62 M 2.3 3 +
14 A 57 M 0.5 5 +

A: atypical carcinoid, T: typical carcinoid, M: male, F: female, +: present, -: absent

Table 2.
The results of analysis of EGFR PNA-mediated clamping using real-time PCR
Sample No(Ct) G719X (①) E19 del. (②) T790M (③) S768I (④) Ins. 3 dup (⑤) Ins.3 (⑥) L858R & L861Q(⑦) Non-PNA (⑧)
15 39.11 38 38 33.24 39.02 36.5 38 28.63
Clamping control 38 36.16 39.79 31.82 30.42 35.92 34.42 23.66
standard Ct 36 33 33 30 27 30 33  
△Ct –2 –0.06 –0.37 –3.91 –3.45 –8 –6.62  
determine wild wild wild wild wild wild Wild  

Criterion: EGFR mutation is noted when the △Ct value, standard Ct minus sample Ct, is 2.0 or above. [△Ct = standard ct – sample ct]

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