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The relationship between the presence of chromosomal instability and prognosis of squamous cell carcinoma of the lung: fluorescence in situ hybridization analysis of paraffin-embedded tissue from 47 Korean patients.

Yoo JW, Seo KW, Jang SJ, Oh YM, Shim TS, Kim WS, Lee DS, Lee SD, Choi CM - J. Korean Med. Sci. (2010)

Bottom Line: Survival rates were compared in terms of age, T factor, N factor, CIN, and smoking status.In multivariate analysis, the presence of CIN was a predictive factor for survival.CIN-positive based on FISH can be prognostic factor of lung SCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary and Critical Care Medicine, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea.

ABSTRACT
To evaluate the prognostic importance of chromosomal instability (CIN) in squamous cell carcinoma (SCC) of the lung, the relationship between CIN detected by fluorescence in situ hybridization (FISH) and survival in SCC patients was examined. Forty-seven surgical specimens of lung SCC were analyzed. To identify tumors with CIN, p16 and multi-target DNA FISH assays for c-myc, chromosome 6, EGFR, and chromosome 5 (LAVysion, Vysis) were performed on nuclei extracted from paraffin-embedded tumor tissues. Survival rates were compared in terms of age, T factor, N factor, CIN, and smoking status. A sample was defined as CIN-positive if at least four of the five chromosomes were positive. Among the 47 specimens, 9 (19%) were CIN-positive. The overall survival rate was 66%. Overall survival rates were estimated as 33.3% for CIN-positive patients and 76.7% for CIN-negative patients (Hazard ratio 3.47; 95% Confidence interval, 1.25-9.67; P=0.017). In multivariate analysis, the presence of CIN was a predictive factor for survival. CIN-positive based on FISH can be prognostic factor of lung SCC.

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Image of fluorescent in situ hybridization (FISH) performed on extracted nuclei of lung cancer tissue. (A) LaVysion FISH: cell showing increase copy number of 7p12, chromosome 5 and 8q24 region. A Cell shows five red signals (7p12 region), three Gold signals (8q 24 region), four green signals (5p15.2) and two aqua signals (chromosome 6). (B) p16 FISH: cell with deletion p 16 gene. A cell shows homozygous deletion of p16 gene (no orange signal) on normal number of chromosome 9 (two green signal).
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Figure 1: Image of fluorescent in situ hybridization (FISH) performed on extracted nuclei of lung cancer tissue. (A) LaVysion FISH: cell showing increase copy number of 7p12, chromosome 5 and 8q24 region. A Cell shows five red signals (7p12 region), three Gold signals (8q 24 region), four green signals (5p15.2) and two aqua signals (chromosome 6). (B) p16 FISH: cell with deletion p 16 gene. A cell shows homozygous deletion of p16 gene (no orange signal) on normal number of chromosome 9 (two green signal).

Mentions: FISH was performed according to the manufacturer's instructions with minor modifications, as described in a previous report (9). The commercially available multi-target FISH assay (LAVysion; Vysis; Downers Grove, IL, USA) includes directly labeled DNA FISH probes for EGFR (7p12,SpectrumRed), MYC (8q24,SpectrumGold), chromosome 5 (5p-15.2,SpectrumGreen), and chromosome 6 (centrometric at 6p11.1-q11,SpectrumAqua). FISH analysis of p16 was conducted using the LSI® p16 (9p21) SpectrumOrange/CEP®9 SpectrumGreen™ Probe (Vysis; IL, USA). After staining, 4',6-diamidino-2-phenylindole (DAPI II) (Abott/Vysis) was applied to the target areas, and the slides analyzed under a fluorescence microscope using single bandpass filter sets. FISH signals were measured at a magnification of ×630 after automatic relocation of equivocal cells with the appropriate software in the DAPI II single bandpass filter set on the microscope. Nuclear signals were only measured in cells with clearly defined nuclear borders and visible signals. In total, 200 nuclei were counted, and the level of each centromeric signal recorded. EGFR, MYC, chromosome 5, and chromosome 6 (Fig. 1) were analyzed for increased copy number, and the p16 gene examined for deletion. Cells displaying gain (>2) or loss (<2) in copy number were determined when they constituted 10% of the total cell population. This cutoff value was based on earlier studies reporting that the mean+3 S.D. value of gain or loss of each chromosome in lymphocytes and normal colorectal epithelium was less than 10% (10).


The relationship between the presence of chromosomal instability and prognosis of squamous cell carcinoma of the lung: fluorescence in situ hybridization analysis of paraffin-embedded tissue from 47 Korean patients.

Yoo JW, Seo KW, Jang SJ, Oh YM, Shim TS, Kim WS, Lee DS, Lee SD, Choi CM - J. Korean Med. Sci. (2010)

Image of fluorescent in situ hybridization (FISH) performed on extracted nuclei of lung cancer tissue. (A) LaVysion FISH: cell showing increase copy number of 7p12, chromosome 5 and 8q24 region. A Cell shows five red signals (7p12 region), three Gold signals (8q 24 region), four green signals (5p15.2) and two aqua signals (chromosome 6). (B) p16 FISH: cell with deletion p 16 gene. A cell shows homozygous deletion of p16 gene (no orange signal) on normal number of chromosome 9 (two green signal).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2877246&req=5

Figure 1: Image of fluorescent in situ hybridization (FISH) performed on extracted nuclei of lung cancer tissue. (A) LaVysion FISH: cell showing increase copy number of 7p12, chromosome 5 and 8q24 region. A Cell shows five red signals (7p12 region), three Gold signals (8q 24 region), four green signals (5p15.2) and two aqua signals (chromosome 6). (B) p16 FISH: cell with deletion p 16 gene. A cell shows homozygous deletion of p16 gene (no orange signal) on normal number of chromosome 9 (two green signal).
Mentions: FISH was performed according to the manufacturer's instructions with minor modifications, as described in a previous report (9). The commercially available multi-target FISH assay (LAVysion; Vysis; Downers Grove, IL, USA) includes directly labeled DNA FISH probes for EGFR (7p12,SpectrumRed), MYC (8q24,SpectrumGold), chromosome 5 (5p-15.2,SpectrumGreen), and chromosome 6 (centrometric at 6p11.1-q11,SpectrumAqua). FISH analysis of p16 was conducted using the LSI® p16 (9p21) SpectrumOrange/CEP®9 SpectrumGreen™ Probe (Vysis; IL, USA). After staining, 4',6-diamidino-2-phenylindole (DAPI II) (Abott/Vysis) was applied to the target areas, and the slides analyzed under a fluorescence microscope using single bandpass filter sets. FISH signals were measured at a magnification of ×630 after automatic relocation of equivocal cells with the appropriate software in the DAPI II single bandpass filter set on the microscope. Nuclear signals were only measured in cells with clearly defined nuclear borders and visible signals. In total, 200 nuclei were counted, and the level of each centromeric signal recorded. EGFR, MYC, chromosome 5, and chromosome 6 (Fig. 1) were analyzed for increased copy number, and the p16 gene examined for deletion. Cells displaying gain (>2) or loss (<2) in copy number were determined when they constituted 10% of the total cell population. This cutoff value was based on earlier studies reporting that the mean+3 S.D. value of gain or loss of each chromosome in lymphocytes and normal colorectal epithelium was less than 10% (10).

Bottom Line: Survival rates were compared in terms of age, T factor, N factor, CIN, and smoking status.In multivariate analysis, the presence of CIN was a predictive factor for survival.CIN-positive based on FISH can be prognostic factor of lung SCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary and Critical Care Medicine, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea.

ABSTRACT
To evaluate the prognostic importance of chromosomal instability (CIN) in squamous cell carcinoma (SCC) of the lung, the relationship between CIN detected by fluorescence in situ hybridization (FISH) and survival in SCC patients was examined. Forty-seven surgical specimens of lung SCC were analyzed. To identify tumors with CIN, p16 and multi-target DNA FISH assays for c-myc, chromosome 6, EGFR, and chromosome 5 (LAVysion, Vysis) were performed on nuclei extracted from paraffin-embedded tumor tissues. Survival rates were compared in terms of age, T factor, N factor, CIN, and smoking status. A sample was defined as CIN-positive if at least four of the five chromosomes were positive. Among the 47 specimens, 9 (19%) were CIN-positive. The overall survival rate was 66%. Overall survival rates were estimated as 33.3% for CIN-positive patients and 76.7% for CIN-negative patients (Hazard ratio 3.47; 95% Confidence interval, 1.25-9.67; P=0.017). In multivariate analysis, the presence of CIN was a predictive factor for survival. CIN-positive based on FISH can be prognostic factor of lung SCC.

Show MeSH
Related in: MedlinePlus