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flow cytometric analysis for intratumoral heterogenity of DNA content in laryngeal sqamous cell carcinoma

Among studies investigating the expression of DNa content in laryngeal SCC there are conflicting results concerning the potentuial prognostic utility of DNA parameters. Intratumoral heterogeneity in macroscopically different tumor areas has previously been demostrated for DNA content of laryngeal SCC. Also our previous investigation demonstrated heterogeneity for AgNOR in different microscopically defined tumor decided to investigate DNa parameters in different microscopically defined tumor areas of SCCD of the larynx. Thirty-one total laryngectomy specimens that were formalin fixed and paraffin embedded have been investigated. Three distinct areas were analyzed: proliferative and invasive tumor margin and tumor center. Speciments were dewaxed at room temperature, then incubated for 90 min in pepsin at 37 oC. Nuclei ewre stained with propidium iodide. Analysis was performed using a FACS calibur flow cytometer( becton Dickinson), using an excitation wavelenght of 488nm and a 15mW argon ion laser ; 20 000 thousand nuclei were counted for each sample.We performed DNA histogram analysis using ModFitLT softwere. DNa ploidy, DNA index, and S-phase fraction werw determined. A significant difference was observed between the inveswtigated atreas concerning the percentage of aneuploid samples( centra:proliferative margin:invasive margin=78:60:40) as well as S phase fraction( p=0.03). Our results, together with our previous data for AgNOR, strongly underline the fact that intratumoral heterogeneity is not to be neglected. The prognostic potential of this approach has yet to be elucidated.

flow cytometry; laryngeal SCC

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