1. Fluorescent In Situ Hybridization (FISH) to Identify Genetic Changes in Fine Needle Biopsy of Lung Lesions Prepared by Jin Jen NCI

2. Why the Need?
Spiral CT screening is highly sensitive for detecting lung cancers in patients at high risk.
Spiral CT plus fine needle biopsy increases the diagnostic rate of accurate diagnoses but a portion of the cases are undetermined.
Molecular markers might be used to assist clinical diagnosis by detecting the presence of specific chromosomal amplifications.

3. How it is Possible? Tumors are highly genetically unstable.
Tumor related changes often are reflected at the chromosome level and are, by definition, specific to tumors.
Genetic analyses can be done in very small tissues and are not subject to the method of detection.

4. Minimal Chromosome Probes That Can Detect Most Lung Cancers (n = 84)
Amplifications
# Detectable by Based on Any Alteration (n =84)
Percentage
1q, 3q, 5p, 8q 83
98.8%
1q, 5q, 8q 80
95.2%
1q, 3q, 8q 78
92.3%
1q, 3q, 5p 77
91.7%
3q, 5p, 8q 75
89.3%

5. Method to Identify Genetic Changes in FNAs by FISH
Identify and prepare BAC probes unique to specific chromosomal regions
Protease digestion of biopsy samples and isolate nuclei
Hybridization with fluorescently labeled BAC probes
Visualize under fluorescent microscope for Ch. copy number changes
Because of the limited time, I will only introduced to one such analysis called … It is basically a method that detects and allows for direct visualization of chromosome number changes in single cells. The general protocol for FISH is list here which involves the ………
Correlate copy number changes with
cytological/pathological findings and clinical outcome

6. Detecting Lung Cancer in FNA-obtained Spiral CT Identified Lung Lesions
Probes Labeling: 1q and 5q (green)
8q and 3q (red)
nuclei (blue)
Samples: 20 paraffin embedded FNA samples formalin fixed biopsies.
All samples have pathology and clinical information.
20 malignant
10 benign specific
10 benign non-specific

7. Sample 503 (Normal Lung) Green: 5p Red: 8q
Normal samples often show 2 signals/cells representing
their diploid status.

8. Sample 577 (Normal Lung) Green: 5p, 2 signals/cell
Red: 8q, 2 signals/cell

9. Cornell Sample: JJ001 Green: 1q, 5 signals Red: 3q, 4 signals
Green: 5p, 9 signals

10. Sample: JJ003 Green: 1q, 6 signals Red: 3q, 4 signals
A case is considered neoplastic or tumor when 5 or more signals were observed in a cell by one or more probes.

11. Sample: JJ005
Green: 1q, 8 signals
Red: 3q, 12 signals

12. Sample JJ006 Green: 1q, 4 signals Red: 3q, 5 signals
Green: 5p, 4 signals
Red: 8q, 5 signals

13. Sample: JJ010
Red: 8q, 6 signals
Green: 5p, 5 signals

14. Sample: JJ011
Green: 1q, 2 signals
Red: 3p, 6 signals

15. Sample: JJ012 Green: 1q, 2 signals Red: 3p, 2 signals
Green: 5p, 2 signals
Red: 8q, 2 signals

16. Sample: JJ014
Green: 1q, >7 signals
Red: 3p, 2 signals

17. Sample: JJ017
Green: 5p, 10 signals
Red: 8q, 7 signals

18. Sample: JJ018 Green: 5p, 3 signals Red: 8q, 6 signals