Cellular apoptosis susceptibility gene expression in endometrial carcinoma:: Correlation with Bcl-2, Bax, and caspase-3 expression and outcome

Deregulation of proliferation and apoptosis is known to contribute to neoplastic transformation and growth. Using specific antibodies for the cellular apoptosis susceptibility (CAS) gene, caspase-3, Bcl-2, and Bax, we examined the protein expression in 89 endometrial carcinomas and in 56 samples of nonneoplastic adjacent endometrium for comparison. Immunostaining results were scored with regard to approximate percentage of positive tumor cells (< 10%, 10% to 50%, > 50%) and relative immunostaining intensity (1+, 2+, 3+). In nonneoplastic endometrium, CAS protein was expressed in 70.6%, Bax in 64%, caspase-3 in 52%, and Bcl-2 in 87%. In neoplastic tissue, CAS was present in 93% of the tumors, Bax in 88%, caspase-3 in 77%, and Bcl-2 in 51%. Bcl-2:Bax ratio was >1 in 9 cases (10%). In cases of atrophy (n=24) and simple (n=10) and complex (n=22) hyperplasia in the adjacent endometrium, lower levels of expression compared with carcinoma were observed for CAS (p=0.003), caspase-3 (p=0.034), and Bax (p=0.04) and higher levels for Bcl-2, although for this protein the results were not statistically significant (p=0.32). There was no association between immunoscores and FIGO stage. High caspase-3 levels were seen in endometrioid tumor type (p=0.017). CAS expression was higher in grade 3 tumors (p=0.002) and older patients (p=0.013). All tumors of younger patients (< 50 years) were Bcl-2 negative (p=0.037). Caspase-3 correlated positively with CAS (p=0.008), Bax (p=0.04), and low Bcl-2:Bax ratio (p=0.043), and inversely (as a trend) with Bcl-2 (p=0.056). Survival analysis (Kaplan-Meier and Cox regression) established a strong association between prognosis and stage, grade, and histologic type (all p less than or equal to0.0036). In addition, shorter survival was observed for patients whose tumors contained >50% of positive cells for caspase-3 (p=0.024) or for CAS (p=0.04). Age, Bcl-2, Bax, and Bcl-2:Bax ratio did not provide prognostic information. Our results suggest a role of CAS, Bcl-2, Bax, and caspase-3, which are apparently involved in the progressive deregulation of proliferation and apoptosis leading from simple and complex hyperplasia to carcinoma. In addition, CAS and caspase-3 protein levels may be useful markers in predicting the outcome of the patients.