Cell Cycle Analysis; DNA Ploidy and S Phase
Test Code
PLOIDY
Associations
Breast cancer, Non-small cell lung carcinoma, Bladder cancer
Methodology
Flow cytometry analysis
CPT Codes
88182 – Flow cytometry, DNA or cell cycle analysis
Turnaround Time
3 days
Specimen Requirements
- 4.0 mL (min. 2.0 mL) whole blood sodium heparin preferred, EDTA whole blood accepted
- 3.0 mL (min. 1.0 mL) bone marrow aspirate sodium heparin preferred, EDTA bone marrow accepted
- 5 mm^3 fresh or frozen tissue in MPLN RPMI transport media
- Formalin fixed paraffin embedded tissue
- 30.0 mL (min 15.0 mL) fresh bladder wash in sterile container
Specimen Stability
- Whole blood or bone marrow stable at 18-25°C for up to 48 hours
- Fresh tissue stable at 2-8°C up to 72 hours
- Frozen tissue stable indefinitely at -20°C
- Formalin fixed paraffin embedded tissue stable at room temperature indefinitely
- Fresh urine stable at 2-8°C up to 48 hours
Storage & Handling
- Whole blood, bone marrow and formalin fixed paraffin embedded tissue, ship ambient
- Fresh tissue and urine, ship in a Styrofoam container with an ice pack (Do not allow the ice pack to directly contact the sample)
- Frozen specimens should be shipped on dry ice.
Causes for Rejection
Insufficient volume; Fresh samples not sent within 24 hours
Reference Range
Diploid population detected
Description
Deoxyribonucleic acid (DNA) ploidy and cell cycle analysis is a rapid and efficient way to evaluate the DNA content (ploidy) and proliferative activity (cell cycle/S-phase fraction) of cells. By staining the DNA with a fluorescent dye, flow cytometry can measure the dye fluorescence in many individual cells, and the data can be analyzed for ploidy (diploid/normal content or aneuploid/abnormal content) and proliferative activity.
Assessment of DNA ploidy in tumors is achieved by determining if an aneuploid peak is present and calculating the DNA index (DI), which is the ratio of the mean tumor sample G0/G1 DNA content compared to that of normal diploid reference cells. Diploid cells (normal or neoplastic) have a DI of 1.0. Aneuploid populations have a DI greater or less than 1.0. The S-phase fraction (SPF), or % S-phase, refers to the percent or proportion of cells preparing for mitosis by active synthesis of DNA. In general, a high SPF correlates with high proliferative activity.
Ploidy and S-phase fraction have been reported to be associated with the grade of neoplastic processes and may have prognostic value. Some studies have shown correlation between DNA ploidy, S-phase proliferation and tumor progression, including tumors from prostate, breast and bladder cancer as well as melanoma.
Assessment of DNA ploidy in tumors is achieved by determining if an aneuploid peak is present and calculating the DNA index (DI), which is the ratio of the mean tumor sample G0/G1 DNA content compared to that of normal diploid reference cells. Diploid cells (normal or neoplastic) have a DI of 1.0. Aneuploid populations have a DI greater or less than 1.0. The S-phase fraction (SPF), or % S-phase, refers to the percent or proportion of cells preparing for mitosis by active synthesis of DNA. In general, a high SPF correlates with high proliferative activity.
Ploidy and S-phase fraction have been reported to be associated with the grade of neoplastic processes and may have prognostic value. Some studies have shown correlation between DNA ploidy, S-phase proliferation and tumor progression, including tumors from prostate, breast and bladder cancer as well as melanoma.
References
- Da Costa Silva Neto, Jacinto et al. (2008). DNA content analysis of breast lobular invasive carcinoma. Internet Journal of Pathology. Volume 7 Number 2.
- Yildirim-Assaf, Selma et al. (2007). The prognostic significance of determining DNA content in breast cancer by DNA image cytometry: the role of high grade aneuploidy in node negative breast cancer. J Clin Path. 60:649-655.
- Isobe, Hiroshi et al. (2006). Prognostic and therapeutic significance of the flow cytometric nuclear DNA content in non-small cell lung cancer. Cancer. Volume 65, Issue 6; 1391-1395.
- Bol, M G W et al. (2003). Proliferation markers and DNA content analysis in urinary bladder TaT1 urothelial cell carcinomas: identification of subgroups with low and high stage progression risks. J Clin Path. 56(6):447-52.
