Prognostic value of flow cytometric DNA analysis in non-small-cell lung cancer: rationale of sequential processing of frozen and paraffin-embedded tissue.

The objective of this study was to determine the prognostic information provided by flow cytometric DNA analysis in non-small-cell lung cancer. Lung samples of 132 consecutive patients submitted to surgery were prospectively processed. When no aneuploid populations were detected in fresh frozen samples, the process continued as a second step in paraffin-embedded tissue, consuming all the tumor available. The influence of ploidy on the postoperative outcome was studied by both a univariate and a multivariate analysis. Aneuploidy was found in 81 patients (61.4%). Fourteen patients showed no aneuploidy in fresh frozen samples; and only after further analysis in paraffin-embedded tissue was abnormal DNA detected. Overall, the 36-month survival was 69% for the diploid group and 24% for the aneuploid group (p = 0.0006). Including subjects submitted to complete tumor removal (stages I, II, and IIIA) in a multivariate analysis adjusted for TNM stage and histologic type, bearers of aneuploid tumors exhibited a higher risk of relapse (hazard ratio 2.65; CI 95% 1.5-4.66;p = 0.004) or death (hazard ratio 2.17; CI 95% 1.08-4.39;p = 0.032) than patients with diploid tumors. DNA ploidy resulted an independent prognostic factor of survival and tumor relapse in completely resected non-small-cell lung cancer. Sequential analysis of fresh and paraffin-embedded samples can help avoid the bias due to intratumoral DNA content heterogeneity. DNA ploidy could be an useful parameter in any future multifactorial analysis of outcome in such tumors.

Analysis of the prognostic significance of cytosolic determination of CA 125 tumor-associated antigen, carcinoembryonic antigen and squamous cell carcinoma antigen in patients with nonsmall cell lung carcinoma.

BACKGROUND: The use of new prognostic factors as guidelines for the indication of treatment ancillary to surgery has been advocated for patients with nonsmall cell lung carcinoma (NSCLC). This article is an analysis of the prognostic information derived from determination of tumor-tissue cytosolic concentration of CA 125 tumor-associated antigen (CA 125), carcinoembryonic antigen (CEA), and squamous cell carcinoma antigen (SCC) in patients with NSCLC. METHODS: Tumor samples were obtained from 97 patients who successfully underwent surgery for NSCLC with curative intent. CA 125 and CEA were determined by enzyme immunoassay. SCC was determined by radioimmunoassay. The relationship between the tumor-tissue marker level and survival or likelihood of disease free survival was analyzed. RESULTS: Thirty-month survival post-treatment in patients registering CA 125 levels less than 26 U/mg was 68%, versus 30% among patients with levels of CA 125 greater tha 26 U/mg (P < 0.005). For SCC, these values were 57% and 39%, respectively (P = 0.07). No significant difference was seen for CEA (60% versus 44%; P = 0.3). Patients whose tumors had CA 125 levels lower than cutoff recorded a disease free survival rate of 61% versus 29% (P < 0.001); with SCC, likelihood of remaining free of tumor recurrence was 55% versus 34% (P < 0.05). Again, no significant difference was seen for CEA (49% versus 45%; P = 0.5). For CA 125 and SCC, the relationship between marker level and outcome held only with the most favorable histologic types. For CA 125 however, this relationship also held for Stage I tumors (P < 0.005). CONCLUSIONS: Ascertainment of concentrations of CA 125 and SCC in tumor tissue is useful parameter in the determination of postoperative prognosis for patients with NSCLC.

Quantitative analysis of carcinoembryonic antigen, squamous cell carcinoma antigen, CA 125, and CA 50 cytosolic content in non-small cell lung cancer.

BACKGROUND: The cytosolic content of carcinoembryonic antigen (CEA), squamous cell carcinoma (SCC), CA 125, and CA 50 antigens in non-small cell lung cancer (NSCLC) is analyzed in this study. The aim was to ascertain the relationship between tumor marker content and the clinicopathologic aspects of this neoplasm. METHODS: Lung tissue samples were obtained at the time of surgery from 75 patients with NSCLC patients (samples of tumor and unaffected tissue) and 29 subjects with idiopathic pneumothorax. All determinations were performed on cytosols obtained from lung specimens. CEA and CA 125 were determined by enzyme immunoassay, SCC antigen by radioimmunoassay, and CA 50 by fluoroimmunoassay. Tumor marker content was analyzed by TNM stage, histologic type, tumor grade, and number of atypias. RESULTS: The concentration of the four markers was significantly higher in cytosol obtained from neoplastic tissue. Frequency of elevated levels of CEA was higher in adenocarcinoma (87% cases expressing high levels of the marker), SCC antigen in epidermoid carcinoma (65% expressing high levels), and CA 125 in large cell carcinomas (100% expressing high levels). No association was found between TNM stage and cytosol concentration for any of the four markers. CEA exhibited significantly greater concentration in well differentiated tumors, whereas this was true of CA 125 in poorly differentiated tumors. CA 125 content was higher in tumors with more atypia. CONCLUSIONS: Cytosolic quantification of tumor markers may be an adjuvant mechanism to evaluate histologic subtypes of non-small cell lung cancer and identification of tumors with poorly differentiated features.