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Study of tumor infiltrating lymphocytes and transforming growth factor-beta as prognostic factors in breast carcinoma., Marrogi, A. J., Munshi A., Merogi A. J., Ohadike Y., El-Habashi A., Marrogi O. L., and Freeman S. M. , International journal of cancer, 1997 Oct 21, Volume 74, Issue 5, p.492-501, (1997) Abstract

Cytokines and growth factors are powerful modulators of the immune response. Their aberrant expression either by the tumor cells or by the tumor infiltrating lymphocytes confers a selective advantage to the tumor to grow and suppress the cytotoxic activity of the infiltrating lymphocytes. Therefore, analysis of these soluble factors in the tumor microenvironment can provide an insight into the understanding of the tumor behavior and may be used as a prognostic factor. In the present study the nature of the tumor infiltrating lymphocytes (TILs) and cytokine profile was examined in 36 and 19 mammary carcinoma tissues, respectively, by immunohistochemistry and PCR. Phenotypic differences in the number of cytotoxic T lymphocytes (CD8+) and lymphokine activated killer cells (CD16) was observed among TILs when patients with either early disease stage (39% and 46.6%, respectively) or those alive with no residual disease (31% and 52%, respectively) were compared with late stage (9.7% and 22.8%, respectively) or those dead of disease (14.6% and 15.6%, respectively). Furthermore, analysis of the 19 tumor samples for cytokine mRNA expression by RT-PCR revealed the presence of TNF-alpha, IL-10, TGF-beta1, and IL-2. However, semi-quantitative PCR analysis demonstrated TGF-beta1 expression to be significantly higher in patients with a favorable outcome (1.0246 attomoles/micromoles) as compared to patients with a poor prognosis (0.1157 attomoles/micromoles). Our results demonstrate the potential biological significance of certain host factors, particularly TILs and TGFbeta1 expression, on the outcome of breast cancer.

DNA ploidy and proliferating cell nuclear antigen image analysis of peritoneal and pleural effusions. A possible diagnostic role., el-Habashi, A. H., Freeman S. M., el-Morsi B., El-Didi M., Martin A., and Marrogi A. J. , Acta cytologica, 1997 May-Jun, Volume 41, Issue 3, p.636-48, (1997) Abstract

OBJECTIVE: To determine the role of DNA and proliferating cell nuclear antigen (PCNA) image analysis (IA) in enhancing the diagnostic sensitivity of conventional cytology (CC).

STUDY DESIGN: The histopathologic and clinical data on 87 consecutive pleural and peritoneal effusions were used to evaluate the accuracy of CC and DNA IA results.

RESULTS: CC showed a sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 65%, 100%, 100% and 62%, respectively. Aneuploidy peaks were seen in 49 cases; 47 of them were true positives. Thirty of 38 diploid cases were true negatives. The sensitivity, specificity, PPV and NPV were 85%, 94%, 96% and 80%, respectively. There were positive correlations between DNA ploidy profile and PCNA proliferative index (PI), (R = .697) and significant differences in PCNA PI between malignant and benign effusions (P < .001).

CONCLUSION: The DNA IA PI by PCNA can be used as a complementary diagnostic tool with CC in cytologically inconclusive cases.

p53 and PCNA coexpression of 81 pleural and peritoneal effusion specimens: an immunohistochemical study., el-Habashi, A. H., Freeman S. M., el-Morsi B., Morris G. F., and Marrogi A. J. , Pathology, research and practice, 1996 Aug, Volume 192, Issue 8, p.834-9, (1996) Abstract

Several studies have indicated that wild type p53 plays an important role in controlling cell growth and acts as a cyclin modifier. Abnormalities in p53 induce the overexpression of proliferating cell nuclear antigen. The aim of this study is to correlate immunocytochemically the expression of mutant p53 and the proliferative index (PI) as indicated by image analysis of PCNA immunoreactivity in 81 cases of pleural and peritoneal effusions. There was a strong correlation (r = 73%) between p53 immunoreactivity and PCNA PIs. Forty-three (71%) cases indicated p53 immunostaining out of 61 cases with PCNA immunoreactivity, forty of which (93%) proved to have a diagnosis of malignancy using histological or clinical data. Furthermore, 7 malignant cases showed PCNA reactivity but no p53 immunostaining. An additional four malignant cases indicated no reactivity for either p53 or PCNA. Also, there was a significant difference in the PCNA PI between benign and malignant effusions (p < 0.001). These clinical observations confirm the function of wild p53 as a check point during cell cycling, and as a strong negative feedback effect on PCNA expression. Furthermore, such co-expression represents a significant indicator for malignancy.