J Korean Med Sci.  2017 Nov;32(11):1784-1791. 10.3346/jkms.2017.32.11.1784.

GAB2 Amplification in Squamous Cell Lung Cancer of Non-Smokers

• Park YR
• Bae SH
• Ji W
• Seo EJ
• Lee JC
• Kim HR
• Jang SJ
• Choi CM

• KMID: 2390304
• DOI: http://doi.org/10.3346/jkms.2017.32.11.1784

Abstract

Lung squamous cell cancer (SCC) is typically found in smokers and has a very low incidence in non-smokers, indicating differences in the tumor biology of lung SCC in smokers and non-smokers. However, the specific mutations that drive tumor growth in non-smokers have not been identified. To identify mutations in lung SCC of non-smokers, we performed a genetic analysis using arrays comparative genomic hybridization (ArrayCGH). We analyzed 19 patients with lung SCC who underwent surgical treatment between April 2005 and April 2015. Clinical characteristics were reviewed, and DNA was extracted from fresh frozen lung cancer specimens. All of copy number alterations from ArrayCGH were validated using The Cancer Genome Atlas (TCGA) copy number variation (CNV) data of lung SCC. We examined the frequency of copy number changes according to the smoking status (non-smoker [n = 8] or smoker [n = 11]). We identified 16 significantly altered regions from ArrayCGH data, three gain and four loss regions overlapped with the TCGA lung squamous cell carcinoma (LUSC) patients. Within these overlapped significant regions, we detected 15 genes that have been reported in the Cancer Gene census. We also found that the proto-oncogene GAB2 (11q14.1) was significantly amplified in non-smokers patients and vice versa in both ArrayCGH and TCGA data. Immunohistochemical analyses showed that GAB2 protein was relatively upregulated in non-smoker than smoker tissues (37.5% vs. 9.0%, P = 0.007). GAB2 amplification may have an important role in the development of lung SCC in non-smokers. GAB2 may represent a potential biomarker for lung SCC in non-smokers.

Keyword

ArrayCGH; Lung SCC; Non-Smoker; GAB2; Proto-oncogene

MeSH Terms

Biology
Carcinoma, Squamous Cell
Censuses
Comparative Genomic Hybridization
DNA
Epithelial Cells*
Genes, Neoplasm
Genome
Humans
Incidence
Lung Neoplasms*
Lung*
Neoplasms, Squamous Cell
Proto-Oncogenes
Smoke
Smoking
DNA
Smoke

Figure

• 

  Fig. 1 Genomic positions of amplified and deleted regions in lung SCC.Statistically significant (A) six amplified and (B) ten deleted regions were found. The X-axis represents the normalized amplification signals (top) and significance by q-value (bottom). The green line represents the significance cutoff at q-value = 0.25.SCC = squamous cell cancer.

• 

  Fig. 2 Frequency of copy number changes for non-smokers and smokers. The frequency of copy number changes was calculated for all measurement points in the arrays and plotted relative to the position along the chromosome for (A) all squamous lung cancer sample, (B) smoker samples, (C) non-smoker samples, and (D) significant copy number differences bewteen smoker and non-smoker. The number of analyzed tumors is indicated. Green bars above the horizontal line indicate the percentage of tumors with copy number gains and red bars below the horizontal line indicate the percentage of tumors with copy number losses.

• 

  Fig. 3 Genetic differences in non-smoking SCC patients compared with smoking SCC patients. X-axis represents genes by chromosomal order, and Y-axis represents 19 samples that are clustered by Euclidian distance based on normalized intensity values. A line plot at the top panel is the P value from a t-test which was log2 transformed.SCC = squamous cell cancer.

• 

  Fig. 4 Immunohistochemical staining for GAB2 in Lung SCC of non-smoker and smoker. (A) Lung SCC of non-smoker was positive for GAB2 staining. The total immunostaining score was 3% and 20% of cancer cells were stained. (B) Lung SCC of smoker was negative for GAB2 staining. While some cancer cells were shown weak positive for GAB2 staining, the percentage of stained cancer cell was less than 5%, and the total immunostaining score was 0.SCC = squamous cell cancer.

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